CLINICAL IMMUNOLOGY (CL)
A DELETERIOUS ROLE FOR IFN-GAMMA AND ITS TC1/M1 GENE SIGNATURE IN HUMAN LEISHMANIASIS:
IMPLICATIONS FOR IMMUNOTHERAPEUTIC AND VACCINATION STRATEGIES
JOHAN VAN WEYENBERGH1; GEORGE SOARES2; THEO THEPEN3; RICARDO KHOURI4; STEFAN BARTH5;
6
7
8
9
GILVANEIA SILVA-SANTOS ; JACKSON M COSTA ; ALDINA BARRAL ; MANOEL BARRAL-NETTO .
1,2,4,6,7,8,9.CPQGM-FIOCRUZ, SALVADOR - BA - BRASIL; 3,5.FRAUNHOFER INSTITUT, AACHEN ALEMANHA.
Introduction: We previously demonstrated a deleterious role for IFN-beta in human leishmaniasis, in contrast to
murine leishmaniasis. In addition, an antagonistic effect of IFN-beta upon IFN-gamma bactericidal activity has been
recently shown in human leprosy, another cutaneous pathology caused by an intracellular pathogen. Since we
described a similar IFN-beta/gamma antagonism for CD64 (FcgammaRI) expression, we hypothesized that high vs.
low CD64 expression, reflecting IFN-gamma vs. IFN-beta activity, respectively, would be associated to a positive vs
negative clinical outcome in human CL.
Methods and Results: We found that ex vivo expression of CD64 in monocytes was significantly elevated in CL.
Contrary to our hypothesis, CD64 MFI positively correlated to therapeutic failure in two independent cohorts
(p=0.0018 and p=0.006). In a prospective study with 110 patients, systemic IFN-gamma levels also significantly
correlated to therapeutic failure (p=0.0013). To further explore a possible IFN-gamma gene signature in human CL
and its relationship to parasite burden in vivo, we employed nCounter technology to quantify >700 human and
selected Leishmania RNAs in situ in skin biopsies taken from the whole clinical spectrum of human CL: uninfected
skin, localized skin lesions (caused by L. braziliensis) and biopsies from patients with incurable diffuse CL (caused by
L. amazonensis). CD8 and GzmB mRNA levels (normalized to G6PD mRNA) significantly correlated to IFN-gamma
RNA in situ, reinforcing our previous finding of a human leishmaniasis dichotomy: CD8/Tc1 for tissue destruction vs.
CD4/Th1 for parasite killing. IFN-gamma mRNA level was also significantly and positively correlated to macrophage
CD64, CD80, CD14 and HLA-DR mRNAs, demonstrating a robust Tc1/M1 gene signature at the site of infection in
human CL. Next, we found that CD64-targeted immunotoxins selectively induce apoptosis and decrease parasite
survival in Leishmania-infected human macrophages. Finally, we demonstrate short-term immunotoxin treatment in
vivo decreases lesion size, parasite load and inflammation in infected HuCD64-transgenic mice only, but is ineffective
in WT mice used as negative control.
Conclusion: Our results provide proof-of-concept for the therapeutic potential of CD64+ macrophage elimination in
cutaneous leishmaniasis and challenge the clinical relevance of established murine models of Th1/Tc1/M1 boosting
for therapeutic and vaccination strategies in humans.
Financial support: PRONEX
AEROBIC EXERCISE MINIMIZES CISPLATIN-INDUCED ACUTE KIDNEY INJURY BY UP-REGULATION OF HO1
MARIANA YASUE SAITO MIYAGI1; MARILIA CERQUEIRA LEITE SEELAENDER2; LUCAS MACERATESI ENJIU3;
PATRÍCIA ESTLER ROCHA GUILHERME4; MARCUS PISCIOTTANO5; MEIRE HIYANE6; CAROLINE YURI
HAYASHIDA7; VINÍCIUS DE ANDRADE OLIVEIRA8; DANILO CANDIDO DE ALMEIDA9; ANGELA CASTOLDI10;
NIELS OLSEN SARAIVA CAMARA11; MARIANE TAMI AMANO12.
1,6,7,8,9,10,11,12.UNIVERSITY OF SÃO PAULO - DEPARTMENT OF IMMUNOLOGY, SÃO PAULO - SP - BRASIL;
2,3,4,5.UNIVERSITY OF SÃO PAULO - DEPARTMENT OF LIPID METABOLISM, SÃO PAULO - SP - BRASIL.
Cisplatin, a chemotherapy drug used in the treatment of several types of cancer, has a limited use by its side effects
as cachexia and nephrotoxicity. Its use increases TNF levels. Previous studies have shown that aerobic exercises
were able to reduce tumors and to diminish cachexia. However, little is known about the impact of exercise in
cisplatin-induced nephrotoxicity. Our aim was to analyze the role of physical exercise on the protection of cisplatininduced acute kidney injury (AKI). Single i.p. injection of cisplatin 20mg/kg in B6 mice was given on day 0 and
sacrificed on day 4. Mice were divided in sedentary control (CT), exercise control (EX), sedentary cisplatin (CIS), and
exercise cisplatin (EX-CIS) group. Exercise was performed in a treadmill with 1 week of adaptation and 6 weeks of
exercise, until the first day after cisplatin injection. Exercise efficacy was confirmed by higher muscle capillarization in
the exercised groups. We then analyzed the effects of exercise in cachexia and we observed that EX-CIS presented
less weight loss (5.9g±1.3) comparing to CIS (7.1g±0.9). We analyzed nephrotoxicity by urea and creatinine levels in
the serum. CT and EX presented similar levels of urea and an increase was observed in CIS (506.0μg/dL±69.8), while
in EX-CIS they were decreased (318.1μg/dL±179.7). Similar results were observed in creatinine levels confirming that
exercise softened cisplatin-induced AKI. We then investigated inflammatory cytokines and we observed that CIS
presented 10019-fold increase in TNF expression in comparison to CT, while EX-CIS presented 4-fold decrease in
comparison to CIS. IL-10 levels were increased in CIS, and basal level of IL-10 expression in EX-CIS indicated that IL10 might be important to control TNF. IL-6, described to upregulate anti-oxidative stress factors in cisplatin-induced
AKI, presented higher expression in CIS-EX. We further investigated the immune regulation involved in this model
analyzing heme oxigenase (HO) 1 by qPCR, WB and IHC, and we observed a higher expression in CIS-EX when
compared to CIS. Apoptosis was shown to be decreased in EX-CIS group by TUNEL analysis. In conclusion, aerobic
exercise induced IL-6 and HO-1 that diminished TNF expression and consequently the inflammation and apoptosis,
contributing to a less aggressive injury. These data suggest that similar results could be expected in other
chemotherapeutic drugs reinforcing the idea of clinical applications.FAPESP 2011/10349-5
ANTI-INFLAMMATORY EFFECT OF STATIN IN DYSLIPIDEMIC PATIENTS WITH CARDIOVASCULAR DISEASE
TACIANA PEREIRA SANT’ANA SANTOS1; MARIANA DE MENEZES PEREIRA2; ISABELA SILVA OLIVEIRA3;
4
5
ROQUE ARAS JÚNIOR ; AJAX MERCÊS ATTA .
1,2,3.PROGRAMA DE PÓS-GRADUAÇÃO EM IMUNOLOGIA, UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR BA - BRASIL; 4.SERVIÇO DE CARDIOLOGIA, HOSPITAL ANA NERY, UNIVERSIDADE FEDERAL DA BAHIA,
SALVADOR - BA - BRASIL; 5.FACULDADE DE FARMÁCIA, UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR BA - BRASIL.
Introduction. Atherosclerosis is the main cause of cardiovascular diseases as myocardial infarction (MI) and stroke.
In the immunopathogenesis of these diseases, participate atherogenic cytokines (IL-1β and TNF-α) and chemokines
(MCP-1 and IL-8), which are negatively regulated by anti-inflammatory cytokines (IL-10 and TGF-β). On the other
hand, statins are inhibitors of the hydroxyl-3-methylglutaryl co-enzyme A (HMG-CoA) reductase that have lipid
lowering properties and anti-inflammatory effects. The aim of this study was to investigate the serum levels of these
cytokines and chemokines in dyslipidemic patients with a previous history of stroke or MI and treated for dyslipidemia
with statin.Methods and Results. Were investigated 86 dyslipidemic patients treated with statin (21 with a previous
history of stroke and 65 with a previous MI history) and 30 healthy control individuals without statin treatment. The
serum levels of cytokines and chemokines were determined by capture ELISA and their median levels inthese three
groups were compared by the Kruskal-Wallis test followed by the Dunn’s posttest. The median levels of the
atherogenic cytokines and chemokines were differentin the groups (IL-1β, P < 0.0001; TNF-α, P = 0.0004; MCP-1, P =
0.0031 and IL-8, P = 0.0104) and were decreased in the patients with a history of stroke or MI. The serum levels of
IL-10 and TGF-β were also different in these groups (IL-10, P = 0.0013 and TGF-β, P < 0.0001). TGF-βlevels were
higher in stroke and MI groups (stroke = 156.6 pg/mL, MI = 192.4 pg/mL, healthy individuals = 108.5 pg/mL), whereas
their IL-10 levels were decreased (stroke = 5.8 pg/mL, MI = 6.1 pg/mL and healthy individuals = 7.8
pg/mL). Conclusions: Statin inhibits the production of IL-1β, TNF-α, MCP-1 and IL-8 in patients with a previous
history of either stroke or MI. Although statin is able to stimulate the production of TGF-β in these individuals, the use
of statin does not increase their production of IL-10.
Financial support: CNPq and CAPES
ASSOCIATION BETWEEN CIRCULATING LEVELS OF SOLUBLE ENDOGLIN, TRANSFORMING GROWTH
FACTOR BETA 1 AND SOLUBLE TUMOR NECROSIS FACTOR ΑLPHA RECEPTORS AND THE CLINICAL
MANIFESTATIONS OF PREECLAMPSIA
LUIZA OLIVEIRA PERUCCI1; KARINA BRAGA GOMES BORGES2; LETÍCIA GONÇALVES FREITAS3; LARA
CARVALHO GODOI4; PATRÍCIA NESSRALLA ALPOIM5; MELINA DE BARROS PINHEIRO6; ALINE SILVA DE
7
8
9
MIRANDA ; ANTÔNIO LÚCIO TEIXEIRA ; LIRLÂNDIA PIRES DE SOUSA .
1,2,3,4,5,7,8,9.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE
6.UNIVERSIDADE FEDERAL DE SÃO JOÃO DEL REI, DIVINÓPOLIS - MG - BRASIL.
-
MG
- BRASIL;
Introduction: Preeclampsia (PE) is characterized by hypertension and proteinuria. It can be classified according to
the severity or to the onset time of the disease. This study aimed to evaluate the associations between the levels of
soluble endoglin (sEng), transforming growth factor beta 1 (TGF-β1) and tumor necrosis factor alpha (TNF-α) soluble
receptors (sTNF-R1 and sTNF-R2), as markers of TNF-α release, and the clinical manifestations of PE. Methods and
Results: This cross-sectional study was previously approved by local Ethical Committee (0618.0.203.000-10) and
included three groups of women: non-pregnant (n=23), normotensive pregnant (n=21) and preeclamptic (n=43). The
last group was divided in mild (n=12), severe (n=31), early (n=19) and late (n=24) PE. Pregnant women were at third
trimester of gestation. Plasma levels of sEng, TGF-β1, TNF-α, sTNF-R1 and sTNF-R2 were determined by ELISA.
Data are reported as median (25th–75th percentiles) or as mean (standard deviation). sEng levels were higher in
preeclamptic [29.3(16.8-52.3)] than in normotensive pregnant [7.8(4.8-9.4)] and non-pregnant [4.5(3.9-5.1)] (all
p<0.001), and in normotensive pregnant compared with non-pregnant (p<0.001). sTNF-R1 levels were higher in
preeclamptic [3479(3182-4339)] than in normotensive pregnant [3028(2468-3606)] (p=0.014) and non-pregnant [1909
(1649-2182)] (p<0.001), and in normotensive pregnant compared with non-pregnant (p<0.001). sTNF-R2 levels were
higher in preeclamptic [8698(1602)] compared with non-pregnant [6508 (1241)] (p<0.001), but no difference was
found comparing to normotensive pregnant [8931(1277)]. The same parameter was increased in normotensive
pregnant compared with non-pregnant (p<0.001). No differences were found in TGF-β1 levels between the three
studied groups. Late PE had higher levels of sTNF-R1 (p=0.004) and sTNF-R2 (p=0.003) than early PE and no
differences were found for sEng and TGF-β1 levels. sEng levels were higher in severe PE than in mild PE (p=0.005)
and no difference was found for TGF-β1, sTNF-R1 and sTNF-R2 levels. Conclusion: Excessive sEng and TNF-α
release in the circulation could play a role in the pathophysiology of PE. In addition, elevated levels of both TNF-α
receptors are associated with late PE, while higher levels of sEng are found in pregnant women with severe PE.
Financial support: CNPq, FAPEMIG and CAPES.
BLOCKING KV1.3 CHANNELS INHIBITS TH2 LYMPHOCYTE FUNCTION AND TREATS A RAT MODEL OF
ASTHMA
SHYNY KOSHY1; MUSTAFA A. ATIK2; REDWAN U. HUQ3; MARK R. TANNER4; PAUL C. PORTER5; FATIMA S.
6
7
8
9
10
KHAN ; MICHEAL W. PENNINGTON ; NICOLA A. HANANIA ; DAVID B. CORRY ; CHRISTINE BEETON .
1,2,3,4,5,6,8,9,10.BAYLOR COLLEGE OF MEDICINE,
INTERNATIONAL, LOUISVILLE - ESTADOS UNIDOS.
HOUSTON
-
ESTADOS
UNIDOS;
7.PEPTIDES
Introduction
Allergic asthma is a chronic inflammatory disease of the airways. Despite the effectiveness of bronchodilators and
anti-inflammatory medication, the incidence of asthma is on the increase. The inflammatory infiltrates in the lungs that
are characteristic of asthma affect the structural cells of the airways, leading to airway hyper-responsiveness. Of the
different infiltrating immune cells, T lymphocytes that produce Th2 cytokines play important roles in the pathogenesis
of asthma. These T cells are mainly fully-differentiated CCR7- effector memory T (TEM) cells. Targeting TEM cells
without affecting CCR7+ naïve and central memory (TCM) cells has the potential of treating TEM-mediated diseases,
such as asthma, without inducing generalized immunosuppression. Studies in the last decades have identified the
voltage-gated Kv1.3 potassium channel as a target for preferential inhibition of T EM cells in Th1/Th17-mediated
autoimmune diseases, both in vitro and in vivo. Here, we investigated the effects of ShK-186, a selective Kv1.3
channel blocker, for the treatment of asthma.
Methods and Results
We collected induced-sputum from patients with asthma and from healthy controls. Using patch-clamp
electrophysiology and flow cytometry we showed that lung-infiltrating T cells in patients are TEM cells and express high
levels of Kv1.3 channels. ShK-186 inhibited the activation of peripheral blood T cells in response to an allergen.
Immunization of F344 rats against endotoxin-free ovalbumin, followed by intranasal challenges with ovalbumin
induced airway hyper-reactivity which was reduced by the administration of ShK-186. ShK-186 also reduced lung
fibrosis, assessed by histology, total immune infiltrates in the bronchoalveolar lavage and number of infiltrating
eosinophils, assessed by differential counts. Rats with the ovalbumin-induced model of asthma had elevated levels of
the Th2 cytokines IL-4, IL-5, and IL-13 measured by ELISA in their bronchoalveolar lavage. ShK-186 administration
reduced levels of IL-4 and IL-5 and induced an increase in the production of IL-10. Finally, ShK-186 inhibited the
proliferation of lung-infiltrating ovalbumin-specific T cells.
Conclusions
Our results suggest that Kv1.3 channels represent effective targets for the treatment of allergic asthma.
Funding from the National Institutes of Health and the American Lung Association.
CASTLEMAN'S DISEASE ASSOCIATED WITH HIGH TITERS OF ANTI-RNP
MARCELLO WEYNES BARROS SILVA; LUIS FÁBIO BARBOSA BOTELHO; INGRID BRASILINO MONTENEGRO
BENTO DE SOUZA.
UFPB, JOÃO PESSOA - PB - BRASIL.
Introduction: Castleman's disease (CD) is a rare lymphoproliferative disorder of controversial origin. It is also known as
angiofollicular lymphoid hyperplasia, giant lymph node hyperplasia and follicular linforreticuloma. The main feature is
the excessive growth of B cells, there is often high levels of IL-6. It is not customary to be associated with the
presence of autoantibodies like anti-RNP that have high specificity for Mixed Connective Tissue Disease. Objectives:
Report the rare association of Castleman disease with anti-RNP positive. Methods and results: Male patient, ESO, 52,
white, left cervical lymphadenopathy reports, painless lymph nodes and mobile. Denies fever, weight loss and night
sweats. He underwent a lymph node biopsy that was suggestive of Castleman's disease and negative for the
presence of HHV-8 virus by hybridization "in situ". Laboratory results detected a positive ANA (anti-RNP> 1:3500),
and tested negative for Anti-DNA, Anti-RO, anti-La, anti-Sm and ANCA. Clinically, it showed an autoimmune disease
such as SLE and scleroderma. Thus, the presence of high titers of anti-RNP antibodies with other negatives are
suggestive of Mixed Connective Tissue Disease (MCTD), but the patient had no symptoms of the disease that is also
statistically more frequent in women. Besides the laboratory diagnosis is required at least one of the clinical criteria:
swelling of hands, synovitis, myositis, Raynaud's phenomenon and acrosclerosis for MCTD. Conclusion: Although
rare, Castleman disease associated with autoantibodies is possible, due to expansion of mature B cells which
although not neoplastic may have varying degrees of loss of tolerance to self-antigens. A review of the literature,
based on database as PubMed, showed that there was no similar cases.
CHARACTERIZATION OF POST-TRAUMATIC IMMUNOS¬¬UPPRESSION AND ITS REVERSION; NOVEL
OBSERVATIONS IN MAJOR SURGICAL TRAUMA PATIENTS
NAHIDUL ISLAM1; MICHAEL WHITEHOUSE2; SANCHIT MEHANDALE3; MICHAEL HALL4; ASHLEY BLOM5;
6
7
8
9
GORDON BANNISTER ; JOHN HINDE ; RHODRI CEREDIG ; BENJAMIN BRADLEY .
1,8.REGENERATIVE MEDICINE INSTITUTE, NCBES, SCHOOL OF MEDICINE, NATIONAL UNIVERSITY OF
IRELAND, GALWAY, GALWAY - IRLANDA; 2,3,5,6,9.ORTHOPAEDIC RESEARCH UNIT, AVON ORTHOPAEDIC
CENTRE, SOUTHMEAD HOSPITAL, UNIVERSITY OF BRISTOL, BRISTOL - REINO UNIDO; 4.SHANNON
APPLIED BIOTECHNOLOGY CENTRE, INSTITUTE OF TECHNOLOGY TRALEE, CO.KERRY, IRELAND, TRALEE IRLANDA; 7.SCHOOL OF MATHEMATICS, STATISTICS AND APPLIED MATHEMATICS, NATIONAL UNIVERSITY
OF IRELAND, GALWAY, GALWAY - IRLANDA.
Introduction Major trauma suppresses immunity thereby increasing vulnerability to nosocomial infections even leads
to organ failure and death. We previously characterised post traumatic immunosuppression (PTI) after joint
arthroplasty which was worsened by donor blood transfusion. By contrast, re-infusion of unwashed autologous
salvaged blood retrieved from surgical wound site reversed PTI1,2. Here we further investigated PTI and its reversal by
biomarkers including DAMPs, chemokines, cytokines. We also characterised these in the salvaged blood.
Methods and Results Two cohorts of knee arthroplasty patients were recruited; NSBT (18) received no blood and
ASBT (25) received salvaged blood retrieved from wound site 6 hours post-operatively. Venous blood was taken preoperatively and within 3-7 days post-operatively. Salvaged blood samples were taken from the bag 1 and 6 hours after
surgery. Samples were assayed for cytokines, chemokines and DAMPS. Postoperative and salvaged plasma were
compared with preoperative levels.
In both study groups, postoperative haematological changes were equivalent. Postoperative changes in NSBT
included increased anti- (IL-5) and decreased pro-inflammatory (IL-1β/2/17A,IFN-g,TNF-α) cytokines. By contrast in
ASBT, changes included increase in pro- (IL-1β/2/17A,IFN-g,TNF-α) with decreases in anti-inflammatory (IL4/5/9/10/13) cytokines. Interestingly, wound-blood showed increases, up-to 100-fold, in DAMPs (HMGB-1,HSP27/60/70,S-100A8/A9,α-Defensin,Annexin-A2), 90-fold in IL-6, 10-fold in TGF-β1, moderate in chemokines (IL-8,MCP1,MIP-1α) and IL-9/10/1ra, marginal in IL-2/12p70/17A, IFN-g and TNF-α,and no change in IL-1β. During the 6 hour
collection time, there were no additional increase in DAMPs, but further increases of 10-fold in IL-6, up-to 10-fold in
chemokines, and little in IL-1β/17A,IFN-g,TNF-α.
Conclusions Wound blood contained normal level of IL-1β with high levels of IL-6 and DAMPs consistent with certain
models of sterile ischemia-reperfusion3. Mechanisms whereby re-infused blood reversed immune status involved
triggering by ingredients within wound-blood supplemented or modified during collection. We conceived PTI to have
evolved to restrain tissue-destructive inflammatory cascades in favour of tissue regenerative processes with
consequent abrogation of normal immune responses to nosocomial infections.
Funding IRC, SFI, NHS Trust
Lancet.363:1025-30,2004
TATM. 12(1).1-45,2011
Bull.NYU.Hosp.Jt.Dis.67(2):182-188,2009
CLINICAL AND LABORATORIAL SCREENING OF PRIMARY IMMUNODEFICIENCIES IN RIO GRANDE DO
NORTE: A NEW CHALLENGE HAS BEGUN
JULIETA GENRE1; LUANDA CANARIO DE SOUZA2; KERCIA MONALINE JOVENTINO3; GILBERTO BEZERRA4;
GERARDO CAVALCANTE BARROSO5; RAISSSA ANIELLE SILVA BRANDÃO6; VICTOR CESAR TAVARES DE SÁ
LEITÃO7; SARAH DANTAS VIANA8; CLEIA TEIXEIRA AMARAL9; MARIA DANTAS VERA10; BEATRIZ COSTA
11
12
13
CARVALHO TAVARES ; ANTONIO CONDINO NETO ; VALERIA SORAYA SALES .
1,2,3,4,5,7,8,13.CLINICAL IMMUNOLOGY LABORATORY, FEDERAL UNIVERSITY OF RIO GRANDE DO NORTE,
NATAL - RN - BRASIL; 6,9,10.PEDIATRIC DEPARTMENT, FEDERAL UNIVERSITY OF RIO GRANDE DO NORTE,
NATAL - RN - BRASIL; 11.PEDIATRIC DEPARTMENT, FEDERAL UNIVERSITY OF SÃO PAULO, SAO PAULO - SP
- BRASIL; 12.LABORATORY OF HUMAN IMMUNOLOGY, BIOMEDICAL SCIENCE INSTITUTE, UNIVERSITY OF
SÃO PAULO, SAO PAULO - SP - BRASIL.
Introduction: Primary immunodeficiencies (PID) are congenital disorders caused by genetic defects in components of
the immune system, leading to an increased susceptibility to infection in affected individuals. The clinical presentation
of these disorders frequently shows overlapping signs and symptoms, making difficult their correct identification and
treatment. Performing a clinical screening and laboratorial, molecular and epidemiologic study of PID in the Brazilian
state of Rio Grande do Norte (RN) plays a critical role in the early diagnosis and treatment of these diseases in this
region of the country and is the primary source of information used to define the underlying immunological defect.
Methods and Results: 1) After analysis of medical records dated from 2005 to 2012 in the Service of Allergy and
Immunology at Pediatric Hospital Professor Heriberto Ferreira Bezerra and clinical screening of the pediatric patients
according to the 10 warning signs of PID, 186 children from 3 months to 17 years old were found with history of
recurrent infections suggestive of PID. 2) Following clinical screening, serum samples of 79 children with suspicious of
PID were evaluated through turbidimetry to assess the levels of immunoglobulins IgG, IgA and IgM. This analysis
revealed two possible cases of IgG deficiency, one of IgA deficiency and one case with all three isotype reduced.
3)The immunological profile established by flow cytometry of 48 children with suspected PID showed altered number
of cells in one or more lymphocytic populations when compared to reference values, including cases of reduced
CD3+, CD4+, CD8+, CD16+CD56+ and CD19+ cells. 4)Employing additional tests, such as dihidrorhodamine assay,
real time PCR, TRECS and sequencing, the molecular diagnosis for 8 PID cases were done: ataxia telangiectasia,
chronic granulomatous disease, x-linked agamaglobulinemia and severe combined immunodeficiency.
Conclusion: Early diagnosis of children with PID in RN through clinical screening and the implementation of
laboratorial and molecular protocols for their identification was essential for referral of the patients to specialized care
centers and the prompt initiation of appropriate therapy, preventing significant disease-associated morbidity.
Furthermore, the accurate diagnosis affords the opportunity to provide appropriate genetic counseling to the patients
and their families and participation of the LASID registry.
Financial support: CAPES
CLINICAL ASSOCIATION OF POSTMENOPAUSAL OSTEOPOROSIS PATIENTS WITH IL-22 SERUM LEVELS
PABLO RAMON GUALBERTO CARDOSO1; THIAGO SOTERO FRAGOSO2; MOACYR JESUS BARRETO DE
MELO RÊGO3; ALEXANDRE DOMINGUES BARBOSA4; IVAN DA ROCHA PITTA5; ANGELA LUZIA BRANCO
6
7
PINTO DUARTE ; MAIRA GALDINO DA ROCHA PITTA .
1,3,5,7.UNIVERSIDADE FEDERAL DE PERNAMBUCO, RECIFE - PE - BRASIL; 2,4,6.HOSPITAL DAS CLÍNICAS
DA UNIVERSIDADE FEDERAL DE PERNAMBUCO, RECIFE - PE - BRASIL.
Introduction: Osteoporosis is a skeletal condition that affects millions of people characterized by low bone mass and
increased risk of fractures. The osteoclasts are influenced by pro-inflammatory cytokines and anti-osteoclastogenic
cytokines can suppress its. The IL-22 cytokine has pro and anti-inflammatory properties; some studies suggest that it
is involved in the development of autoimmune diseases and allergic airway inflammation.
Methods and Results: A total of 52 patients with postmenopausal osteoporosis and 18 patients without
postmenopausal osteoporosis were recruited from Clinical Hospital of Federal University of Pernambuco
(UFPE). Bone mineral density (BMD) was measured by dual-energy x-ray absorptiometry (DXA) and used to
diagnose osteoporosis according to the classification of the World Health Organization (WHO). Patients were clinically
evaluated; then, blood samples were collected, centrifuged, and the sera separated and stored at -80 ºC. IL-22 in
serum was assayed with an ELISA kit. IL-22 was observed in a greater number of patients with postmenopausal
osteoporosis. IL-22 in serum of patients with post-menopausal osteoporosis (mean 47,04 pg/ml ± 60,25 pg/ml) and
without post-menopausal osteoporosis (mean 27,24 pg/ml ± 20,99 pg/ml) P = 0,2780. High IL-22 levels was observed
in patients with more advanced disease. The levels of IL-22 are especially high in shorter periods of menopause.
When it is durable, levels of IL-22 is low (P = 0,0289; r² = 0,05773).
Conclusion: We found levels of IL-22 increased in patients with shorter duration of menopause. Since the decrease
in estrogen early in the menopause is higher, there is an increased production of IL-22 at the time of production,
suggesting that this cytokine acting in the first years of the disease.
Financial support: INCT_if; CAPES; FACEPE.
DEFECTS IN IL-12 / IFN-Y AXIS: A NEW PERSPECTIVE ON DEVELOPMENT OF WHEEZING
ANGELA FALCAI1; PAULO VÍTOR SOEIRO-PEREIRA2; CHRISTINA ARSLANIAN3; ISABEL RUGUE GENOV4;
5
6
7
VERA RULLO ; DIRCEU SÖLÉ ; ANTONIO CONDINO NETO .
1,2,3,7.UNIVERSITY OF SÃO PAULO, DEPARTMENT OF IMMUNOLOGY, INSTITUTE OF BIOMEDICAL
SCIENCES, SÃO PAULO - SP - BRASIL; 4,6.FEDERAL UNIVERSITY OF SÃO PAULO MEDICAL SCHOOL, SÃO
PAULO-SP, BRAZIL, DIVISION OF ALLERGY, CLINICAL, SÃO PAULO - SP - BRASIL; 5.SANTOS MEDICAL
SCHOOL, SANTOS - SP, BRAZIL, SÃO PAULO - SP - BRASIL.
Introduction: At least 20% of children up to 2 years old show recurrent wheezing (RW) that has been generally
associated with asthma prognosis.
Aim: Evaluate surface markers, transcription factors and cytokine production in cells from RW children.
Methods: We evaluated 25 RW and 9 non-RW children (around 30 months of age). Surface markers and
transcriptional factors were analyzed by flow cytometry on peripheral blood mononuclear cells (PMBC). Cytokine
production was also assessed in response to in vitro stimulation with interferon gamma (IFN-γ), interleukin 12 (IL-12),
phytohemagglutinin (PHA), lipopolysaccharide (LPS), Dermatophagoides pteronissinus (Der p), Blomia tropicalis (Blo
t) and Bla g (Bla g) allergens. Mann Whitney statistical test.
Results: We observed no differences in cell surface receptors (p>0.05). However, lymphocytes from RW children
expressed higher levels of STAT3 and T-bet compared with non-RW (p<0.05). PBMC from RW children released
lower amounts of IL-12 after stimulation with IFN-γ, LPS and Bla g and produced lower levels of IFN-γ after stimulation
with LPS, Blo t and Bla g compared with non-RW children (p<0.05). It was also observed a decrease in TNF-α release
after stimulation with TLR-3, -5 agonists and Derp by PBMC from RW compared to non-RW (p<0,05).
Conclusion: PBMC from recurrent children presents failures in IL-12/IFN-γ production after specific antigen
stimulation, confirming our previous results that RW children have impairment in Th1 response. We also conclude that
lymphocytes of RW present T-bet and stat-1 overexpression, but no defects in receptor of IL-12R, IFN-yR, CD14 and
TLR4 expression. Our research brings a new perspective to understand the establishment of wheezing and asthma.
Financial support: FAPESP, CNPq
DETECTION AND QUANTIFICATION OF EIGHT HERPESVIRUSES IN PATIENTS UNDERGOING ALLOGENEIC
HEMATOPOIETIC STEM CELL TRANSPLANTATION IN BRAZIL
MIRIAM YURIKA HIRAMOTO UEDA1; JULIANA MONTE REAL2; PAULO GUILHERME ALVARENGA GOMES DE
3
4
5
OLIVEIRA ; ELOISA DE SA MOREIRA ; LUIZ FERNANDO LIMA REIS ; CELSO FRANCISCO HERNANDES
6
7
GRANATO ; CELSO ARRAIS RODRIGUES .
1,3,6,7.UNIVERSIDADE FEDERAL DE SÃO PAULO - UNIFESP, SÃO PAULO - SP - BRASIL; 2,5.HOSPITAL SÍRIO
LIBANÊS, SÃO PAULO - SP - BRASIL; 4.DENDRIX RESEARCH, SÃO PAULO - SP - BRASIL.
Miriam yurika hiramoto Ueda (1,2); Juliana Monte Real (2); Paulo Guilherme ALVARENGA GOMES DE Oliveira (3);
Eloisa de Sa Moreira (4); Luiz Fernando Lima Reis (2); Celso Francisco Hernandes Granato (1); Celso Arrais
Rodrigues (2,3)
- Laboratorio de Virologia, UNIFESP, Sao Paulo, Brazil
- Hospital Sírio-Libanês, Sao Paulo, Brazil
- Disciplina de Hematologia e Hemoterapia, UNIFESP, Sao Paulo, Brazil
- Dendrix Research, Sao Paulo, Brazil,
Introduction: Hematopoietic stem cell transplantation (HSCT) is a potentially curative procedure to many
hematological diseases. Human herpesviruses may cause severe complications after HSCT such as interstitial
pneumonia, encephalitis, delayed engraftment and post-transplant lymphoproliferative disease (PTLD). A prospective
survey on the incidence of primary infection or reactivation and clinical features of herpesvirus infections after HSCT
has not yet been performed in Brazilian patients. Additionally, the impact of most of these infections on the HSCT
outcome is still unclear. This study aimed to develop a test to screen and quantify eight human herpesviruses in
patients undergoing a HSCT.
Methods: From March 2011 to August 2012, peripheral blood samples from 99 HSCT recipients were collected once
a week after transplant until day +100, totalizing 824 samples. In a semi-automated workflow, the DNA was extracted
from plasma in the QIAcube robot. A test based on quantitative real-time PCR (Taqman®) was optimized to screen
and quantify all known human herpesviruses (CMV, EBV, HSV1, HSV2, VZV, HHV6, HHV7 and HHV8). The PCR
reactions were set up using QIAgility robot for high-precision pipetting, and have been performed in a 7900HT (Life
Technologies). Infected cell cultures and plasma specimens with a known viral load/amplicon copy number have been
used as controls.
Results: The limit of detection of the qPCR was 5 copies per reaction, representing 250 copies/mL of plasma for all of
the viruses. The efficiencies obtained vary between 90–100% and linearity ranged from 25 to 108 copies per reaction.
No cross-reaction or false positive results were detected. The incidences of primary infection or reactivation of
herpesviruses were: CMV=24%, HHV6=10%, HHV8=5.5%, EBV=2.7%, HSV1=2.7%, VZV=2.7%, HHV7=1.8%, and
HSV2=0.9%. HHV6 was significantly more common after umbilical cord blood transplant than after adult stem cell
transplant. Considering the impact of those infections in transplant outcomes, CMV had no significant impact, HHV6
was associated with an increased risk of platelet engraftment failure, and HHV8 was related with increased risk of
chronic graft-versus-host disease (GVHD).
Conclusion: CMV and HHV6 primary infection or reactivation are frequent after HSCT and may impact on the
transplant outcomes. Monitoring these viruses constitute an essential measure to improve outcomes.
Financial support: FAPESP.
DEVELOPMENT AND VALIDATION OF MULTIPLEX TEST FOR MEASUREMENT OF ANTIBODIES AGAINST C.
DIPHTERIAE, C. TETANI AND H. INFLUENZAE TIPO B (HIB)
TAMIRIS AZAMOR; ANDRÉA DA SILVA; ALESSANDRO DE SOUZA; LUCIANA TUBARÃO; PATRÍCIA NEVES;
DENISE MATOS.
BIO-MANGUINHOS/ FIOCRUZ, RIO DE JANEIRO - RJ - BRASIL.
Introduction and objectives: Liquid microarray, a microsphere-based Multiplex assay is a method that is replacing
immunosorbent assay (ELISA) in assessing the immunogenicity of multicomponent vaccines in preclinical and clinical
trials. This technology utilizes fluorescent distinct microspheres as carriers for different molecules allowing the
simultaneous detection of multiple reactions in a small volume of sample. Studies have demonstrated multiplex
effectivity in a variety of assays for detection and quantification of antibodies and antigens, with high reproducibility
and sensitivity (J. Immun. Met. 335:79-89, 2008). Therein, the aim of this study was to develop and validate a
monoplex and subsequently the Multiplex assay to quantify IgG antibody against diphtheria and tetanus toxins, and
Hib.
Methods and Results: Purified diphtheria, tetanus and phosphoribosylribitol phosphate (PRRP) capsule from Hib,
(DTHib) antigens were coupled to microspheres with different concentrations of those. Standard curve was prepared
used International Reference Serum from NIBISC, and the concentration for tetanus and diphtheria ranged from
0.13UI/mL to 0.0002 and for PRRP ranged from 0.4 μg/mL to 0.0005μg/mL. By optimizing of the DTHib monoplexes
the best results were obtained with concentrations of 10μg/mL for diphtheria, 1μg/mL for tetanus and 400μg/mL for
PRRP. Twenty sera from human pre and post immunized with DTP/Hib vaccine were analyzed by both ELISA and
monoplex assays under conditions established. Concentrations of IgG antibodies were determined by 4-parameter
logistic standard curve using the SoftMax® program. The values in Mean Fluorescence Intensity (MFI) were converted
to IU/mL or μg/mL, also by interpolation for standard curve (4-parameter) for every microesphere region/standard.
Values obtained by this assay were compared to those found by ELISA. Nonparametric Spearman analysis showed
values statistical significant and good correlation between methods with coefficient of 0.608 (p=0.0045) for diphtheria,
0.775 (p=0.001) for tetanus. PRRP results are still being concluded.
Conclusions: The present study show promising results between ELISA and monoplex methods. From these results
we proceed with the validation of Multiplex assay to evaluate the immunogenicity of the DTP/Hib vaccine.
Financial support: Bio-Manguinhos
DISTINCT EXPRESSION PROFILES OF IL32 ISOFORMS IN MYELODYSPLASTIC SYNDROMES
MATHEUS R LOPES1; FABIOLA TRAINA2; JOÃO KLEBER N PEREIRA3; PAULA DE MELO CAMPOS4; JOÃO
AGOSTINHO MACHADO-NETO5; IRENE LORAND-METZE6; SARA T OLALLA SAAD7; PATRICIA FAVARO8.
1,2,3,4,5,6,7.HEMATOLOGY AND HEMOTHERAPY CENTER - UNIVERSITY OF CAMPINAS, INCT DO SANGUE,
CAMPINAS - SP - BRASIL; 8.DEPARTMENT OF BIOLOGICAL SCIENCES, FEDERAL UNIVERSITY OF SÃO
PAULO, DIADEMA - SP - BRASIL.
Introduction: IL32 has been reported as an important mediator in several autoimmune and inflammatory disorders.
There are six splice variants (α,β,γ,δ,ε,φ). IL32 can induce the production of MIP2, IL1β, IL6, IL8, IL10 and TNFα, the
latter described as a key player in the pathophysiology of MDS. Although little is known about IL32 in MDS, marrow
stroma from MDS patients has been described to present higher levels of IL32 mRNA, suggesting an
autoamplification loop of TNFα and IL32. Herein, we investigated the IL32 and its isoform expressions (α,β,δ,γ) in
peripheral blood CD3+ cells from controls and MDS patients. These data were correlated with CD4+ and
CD8+ lymphocyte profiles and with the expression of FOXP3, IL10, TGFβ1, CTLA4 and TNFα. Methods: 29 samples
from controls and 52 patients with MDS (WHO: low-risk=43, high-risk=09) were included in the study. The study was
approved by the ethics committee. T cell frequencies were determined by flow cytometry. The expression levels from
CD3+ cells were determined by qPCR. The statistical methods used were Mann Whitney test and Spearman
correlation. Results: IL32 mRNA expression in peripheral CD3+ cells showed a decreased expression in MDS
patients when compared with controls (P=0.02) and may be a reflection of the isoform IL32γ, which also presented a
lower expression in MDS patients (P=0.02). This statistical difference remained after we classified the patients into
subgroups, but the decreased IL32γ expression was more pronounced in the high-risk MDS (P=0.02). There was no
statistical difference between the MDS and the control groups regarding the other isoforms studied. IL32 correlated
positively with FOXP3, CTLA4 and TGFβ1 expression (P<0.02, r<0.3) in MDS patients. A similar pattern is observed
between isoform IL32δ and TGFβ1 (P=0.02, r=0.37). Isoform IL32γ transcripts positively correlated with
TNFα mRNA levels (P=0.05, r=0,32). Conclusion: Contrary to our expectations, the production of IL32 in peripheral
CD3+ cells of the MDS patients was significantly lower than in controls. This result, together with the positive
correlation of isoform IL32δ with TGFβ1, indicate that IL32δ may play a role in the downregulation of inflammatory
responses. The correlation of isoform IL32γ with TNFα supports the role of this isoform in the pathophysiology of
MDS. Further studies are necessary to better characterize the function of the IL32 variants in MDS patients.
Supported by FAPESP, CNPq and INCT do Sangue.
DISTRIBUTION OF TH1, TH2, TH17 AND EXPRESSION OF FAS E FASL IN CD4+ T LYMPHOCYTES FROM BCLL PATIENTS
1
2
3
FLAVIA AMOROSO MATOS E SILVA ; RODOLFO PATUSSI CORREIA ; NELSON HAMERSCHLACK ; NYDIA
BACAL4; RODRIGO SANTUCCI5; GUSTAVO PESSINI AMARANTE-MENDES6.
1,2.DEPARTAMENTO DE IMUNOLOGIA, INSTITUTO DE CIÊNCIAS BIOMÉDICAS, UNIVERSIDADE DE SÃO
PAULO, SÃO PAULO - SP - BRASIL; 3,4.HOSPITAL ISRAELITA ALBERT EINSTEIN, SÃO PAULO - SP - BRASIL;
5.HOSPITAL ESTADUAL MARIO COVAS, SÃO PAULO - SP - BRASIL; 6.DEPARTAMENTO DE IMUNOLOGIA,
INSTITUTO DE CIÊNCIAS BIOMÉDICAS, UNIVERSIDADE DE SÃO PAULO; INSTITUTO D, SÃO PAULO - SP BRASIL.
B-CLL is characterized by the progressive accumulation of small B lymphocytes that do not undergo apoptosis due to
an underlying defect. One potential mechanism of defective apoptosis may be irregular cytokine production by T cells.
Our hypothesis is that patient’s CD4+ T cells may participate in the course of this hematological malignancy and
contribute to the pathophysiology and evolution of the disease. In this context, we aimed to determine the cytokine
profile in the serum of B-CLL patients, the relative frequency of TH1, TH2, TH17, as well as the expression of FAS
and FASL in CD4+ T lymphocytes. Patients and Methods: We performed flow cytometry analysis of peripheral blood
mononuclear cell samples from 26 healthy donors and 25 chronic lymphocytic leukemia (B-CLL) patients. TH1, TH2,
TH17 cells were determined by the expression of INF-γ, IL-4 and IL-17, respectively. Results: We observed an
increased frequency of TH1 cells in B-CLL patients compared to healthy donors. Regarding the expression of FAS
and FASL results showed only increased of FASL. Thus each patient was analyzed and compared to the prognostic
factors and to clinical data in each case and hasn’t showed correlation. Conclusion: These results suggested that
IFN-γ-producing TH1 cells and increase expression of FASL in CD4 T lymphocytes could play an important role in the
outcome of B-CLL. Financial support: FAPESP, CNPq and Instituto Israelita de Ensino e Pesquisa Albert Einstein IIEPAE.
EARLY VIROLOGIC RESPONSE AS NEGATIVE PREDICTIVE VALUE FOR SUSTAINED VIROLOGIC RESPONSE
IN PATIENTS WITH CHRONIC HEPATITIS C GENOTYPE 1 NAÏVE.
MARIANA SARTORI MAGNONI1; FERNANDA MACEDO NALLI SILVA2; MARJORIE DE ASSIS GOLIM3; FABIO DA
4
5
6
SILVA YAMASHIRO ; CASSIO DE OLIVEIRA VIEIRA ; PAULO EDUARDO ABREU MACHADO ; EDIPO VINÍCIUS
7
8
ALMEIDA FERREIRA ; SARITA FIORELLI DIAS BARRETO ; REJANE MARIA TOMMASINI GROTTO9; MARIA INÊS
10
11
DE MOURA CAMPOS PARDINI ; GIOVANNI FARIA SILVA .
1,3,6.LABORATÓRIO DE CITOMETRIA DE FLUXO - HEMOCENTRO DE BOTUCATU - UNESP, BOTUCATU - SP BRASIL; 2,4,5,11.DEPARTAMENTO DE CLÍNICA MÉDICA - FACULDADE DE MEDICINA DE BOTUCATU - UNESP,
BOTUCATU - SP - BRASIL; 7,8,9,10.LABORATÓRIO DE BIOLOGIA MOLECULAR - DIVISÃO HEMOCENTRO UNESP, BOTUCATU - SP - BRASIL.
Introduction: Hepatitis C affects approximately 200 milion people in the world, where most progress to the chronic
phase to cirrhosis or hepatocellular carcinoma. Treatment is combination of pegylated interferon with ribarivin (PEGIFN and RBV) to obtain sustained virological response (SVR). Achieving rapid virological response (RVR), has
demonstrated positive predictive value (PPV) for high SVR. Both the early virologic response (ERV) and PRV, has
been shown to not predictive of SVR evolution. The objective was to evaluate SVR during treatment, and thus patients
followed about the factors that may be predictive of achieving SVR or not, considering the degree of fibrosis and
medication. Methods and Results: A retrospective descriptive study was performed using database of the
Department of Interne Medicine Gastroenterology Division, Botucatu School of Medicine, São Paulo State University,
UNESP, Botucatu, SP, Brazil, and patients were included: naïve (treatment-naïve) with HCV- RNA detected genotype
1 (1a, 1b), treatment with Pegintron (MSD®), or Pegasys (Roche®), and not co-infected with HIV / HBV. We included
261 patients 63,2% male and 37,8% female, 31,9%b for age ≤ 40 years and 68,1% for > 40 years. 61,8% degree ≤ F2
fibrosis and 38,2% degree ≥ F3. As the medication 67% and 33% used Pegintron or Pegasys. The RVR was achieve
by 16,1% of subjects, 86% of these progressed to SVR, those who did not achieve RVR 63,8% also did not progress
to SVR. Among individuals who obtained 52,9% PVR, 67,5% of these achieved SVR, in return for that failed PVR
81,2% did nor progress to SVR. Regarding fibrosis, RVR, PVR and SVR were more often observed in patients degree
≤ F2 beign respectively 76,1%, 64,5% and 75,4%. Of those using Pegintron RVR reached 13,8%, 42,5% and PVRSVR 42,5%, in which 25,9% had used pegasys RVR, 56,5% and 45,9% PVR-SVR. In this regard it was noted that the
positive predictive value for RVR is 86% and negative predictive value for PVR is 81,2%. Conclusion: Analyzing the
variables it is concluded that there is a difficulty to achieve SVR when it reaches at in the treatment of hepatitis C.
EFECTS OF PHYSICAL TRAINING ON INFLAMMATORY PROFILE OF PATIENTS WITH OBSTRUCTVE SLEEP
APNEA
EDUARDO ALVES; VALDIR DE AQUINO LEMOS; CAROLINA ACKEL D´ELIA; GILBERTO PANTINGA JUNIOR;
FABIO S LIRA; GABRIELA PONTES LUZ; TTEREZA BARROS SCHÜTZ; NADINE MARQUES NUNES; ROSENY
PEREIRA DE SOUZA; LIA AZEREDO BITTENCOURT; SERGIO TUFIK; MARCO TULIO DE MELLO.
UNIFESP, SAO PAULO - SP - BRASIL.
Introduction: Obstructive Sleep Apnea (OSA) is characterized by recurrent episodes of upper airway obstruction
during sleep causing sleep fragmentation and metabolic changes such as increased pro-inflammatory cytokines [Prog
Cardiovasc Dis. 51(5):392-9, 2009]. Physical exercise training is largely applied as non-pharmacological treatment in
a lot of diseases [Lipids Health Dis. 10:148, 2011] however the effects of physical training on cytokines profile of OSA
patients remain uncertain. The aim of the present study was to evaluate the effect of physical training on inflammatory
profile of patients with OSA.Methods and Results: Six men (mean ± SD of age 48.1 ± 13.9 years, body mass 84.07 ±
6.26 kg; height 172.0 ± 0.07 cm; BMI 28.41 ± 1.60 kg/m 2) diagnosed with moderate OSA were submitted to a
combined physical training (aerobic + resistance training) three times per week for 4 months. Training session
consisted of 5 minutes of stretching, 30 minutes of aerobic exercise and 25 minutes of resistance training. Blood
samples for cytokines analysis (IL-1ra, IL-1, IL-6 e IL-10 e TNFα) were collected in three different moments: baseline
and after two and four months of physical training. Data were analyzed using ANOVA followed by Tukey post-hoc test
and significance level of 0.05 was adopted. It was observed a significant decrease of IL-6 after two (1.7 ± 1.5 pg/ml)
and four months of training (1.3 ± 1.0 pg/ml) compared to baseline condition (19.7± 18.0 pg/ml; p<0.05).Moreover,
after four months of training (23.4 ± 18.7 pg/ml) it was observed a significant decrease of TNF-α compared to baseline
condition (43.3 ± 19.0 pg/ml; p<0.05). Conclusion: Physical training reduces pro-inflammatory cytokines IL-6 and
TNF-α in patients with OSA. These results suggest that physical training can promote anti-inflammatory effects in this
population.
Financial Suport: CEPE, CEMSA, AFIP, CNPq and FAPESP
EFFECTIVENESS OF ANTIMICROBIAL AND IMMUNOMODULATORY EFFECT OF BACTERIOPHAGES IN
GENITAL DISEASES OF WOMEN
ANATOLIY P. GODOVALOV1; LILIYA P. BYKOVA2; TATYANA YU. DANIELYAN3; NARINE A. DANIELYAN4.
1,2.ACAD. E.A. WAGNER PERM STATE MEDICAL ACADEMY, PERM - RÚSSIA; 3,4.MEDITSINSKAYA STUDIYA
LLC, PERM - RÚSSIA.
Introduction: The use of bacteriophages in the treatment of inflammatory diseases of the genital area is of increasing
interest. Highly-valued property of bacteriophages is the narrow specificity of effect. In addition to direct antimicrobial
effect phages have immunomodulatory properties, they increase the phagocytic activity of leukocytes, enhance the
respiratory action of leukocytes, stimulate the production of cytokines and generation of antibodies to non-phage
antigens (Gorski et al., 2012). The result may be elimination of bacteria from the inflammation area.
Aim. The aim of the study is to evaluate the clinical and microbiological effects of immunomodulatory effect of
polyvalent bacteriophage in treatment of inflammatory diseases of genitals.
Methods: 36 patients in Group 1 got a combined treatment with antibiotics and polyvalent bacteriophage "Sekstafag"
(5 ml was used for irrigation and 15 ml - orally for 10 days). 27 patients in Group 2 were treated with antibiotics for
comparison. The effectiveness of treatment was evaluated clinically and with bacteriological method. Statistical
processing of the data was performed using Student t-test.
Results. Chronic endometritis was identified in 83%, chronic salpingo-oophoritis in 88%, and chronic cervicitis - in
68% of cases. From the cervical canal of the patients we secreted St. aureus in 42%, St. intermedius in 16%, St.
hyicus in 16%, St. haemolyticus in 10% of cases. 100% of St. hyicus, 87% of St. aureus, 67% of St. intermedius, 50%
of St. haemolyticus were phage-sensitive. Phage-sensitive E. coli were secreted from cervical canal of 21% of women.
Phage therapy has led to the disappearance of pain symptoms in 89% of women of the main group and in 74% of
patients in comparison group. After 7 days of treatment emissions from genital tracts decreased in 94% of patients of
the main group and in 67% of the comparison group. “Sekstafag” therapy resulted in the disappearance of St. aureus,
St. hyicus, Kl. pneumoniae, E. coli, Enterobacter sp. Cases of Lactobacillus sp. secretion after phage therapy
increased from 5% to 32% (p<0.05). Immunomodulatory effect of bacteriophage has also contributed to the
elimination of Candida representatives non-specific to phage (p<0.05).
Conclusion. The conducted study showed the high-effectiveness of suggested treatment with use of bacteriophage.
EOSINOPHILIA IN ADULT PATIENTS WITH ATOPIC DERMATITIS IS ASSOTIATED WITH AN IMPAIRED OF
CD38 AND CD69 EXPRESSION
TIAGO DE OLIVEIRA TITZ; VANESSA GONÇALVES DOS SANTOS; CAMILA DE LOLLO; RAQUEL LEÃO ORFALI;
ALBERTO JOSE DA SILVA DUARTE; MARIA NOTOMI SATO; VALERIA AOKI.
FACULDADE DE MEDICINA, SÃO PAULO - SP - BRASIL.
Introduction and Objectives: Atopic dermatitis (AD) is a chronic recurrent inflammatory disease, with prevalence
around 10 to 20% in children and 1-3% in adults and its diagnosis is based on clinical features. Eosinophils, a
multifunctional polymorphonuclear leukocytes are implicated in the pathogenesis of numerous inflammatory allergic
processes including atopic dermatitis. The aim of this study was to evaluate activation markers expression on
eosinophils from peripheral blood and correlate with disease severity.
Methods and Results: This work enrolled 12 patients with AD from Hospital das Clínicas- São Paulo, (according to
diagnostic criteria of Hanifin & Rajka), 2 men and 10 women, aged between 21 and 61, and 20 healthy controls, aged
between 24 and 53 years, 8 men and 13 women. The severity of the disease was established according to EASI
(Eczema Area and Severity Index, patients graded as mild (58,3%), moderate (16,6%) and severe (25%). Eosinophils
(Lineage cocktail 1- CCR3+) from peripheral blood were analyzed for CD38, CD69, CD23 and CD62L, by flow
cytometry (LSRFortessa, BD Biosciences) and analysis was performed using the FlowJo 7.5.6 software. Our findings
reveal an increased eosinophils CCR3+ percentage and upregulation of CCR3 medium fluorescence intensity (MFI) in
AD individuals compared to controls. Moreover, the early and late activation markers, CD69 and CD38, respectively,
in percentages are both increased in AD patients, while the expression was markedly decreased. No difference of
CD23 and CD62L expressions on eosinophils were detected in the analyzed groups. All findings were not associated
with severity of the disease.
Conclusion: Eosinophilia is a hallmark in atopic dermatitis, whereas is associated with decreased activation of CD38
and CD69 markers. The eosinophil functional parameters needs to be further evaluated in the Atopic Dermatitis.
Financial support: FAPESP/LIM-56/HCFMUSP.
ETHANOL-INDUCED GASTRIC INJURY: MICROSCOPIC ANALYSIS OF THE PROTECTIVE EFFECT OF
FRUTALIN
1
2
LUANA TORRES MONTEIRO MELO ; ANA PAULA VASCONCELLOS ABDON ; RENATA PRADO
VASCONCELOS3; CAROLINA ARAÚJO CASTRO4; MARJORIE MOREIRA GUEDES5; ADRIANA DA ROCHA
TOMÉ6; ANDRÉ LUIZ HERZOG CARDOSO7; THIAGO DE MELO SANTIAGO8; LUCIANA MAGALHÃES REBÊLO9;
RENATO DE AZEVEDO MOREIRA10; ANA CRISTINA DE OLIVEIRA MONTEIRO-MOREIRA11; ADRIANA ROLIM
CAMPOS12.
1,2,3,4,10,11,12.UNIVERSITY OF FORTALEZA (UNIFOR), FORTALEZA - CE - BRASIL; 5.FEDERAL UNIVERSITY
OF CEARÁ, FORTALEZA - CE - BRASIL; 6,7,8,9.STATE UNIVERSITY OF CEARÁ, FORTALEZA - CE - BRASIL.
Obtained from breadfruit seeds (Artocarpus incisa), frutalin (FTL) has a range of important pharmacological
properties. FTL activates and modulates lymphocytes and neutrophils and possesses gastroprotective effects. The
purpose of this study was to evaluate the ability of FTL to protect the gastric mucosa of mice submitted to
ethanolinduced gastric injury. The gastroprotective effect of FTL was evaluated using a murine model of ethanolinduced gastric injury. Damage of the gastric mucosa was assessed morphologically by light, scanning electron and
atomic force microscopy. Lymphocyte infiltration, necrosis, disruption of the mucosal structures and epithelial
desquamation were observed in the control group. In the group pretreated with FTL at 0.5 mg/kg, gastroprotection,
reduced tissue damage and preserved submucosal and mucosal structures were observed. Our findings confirm
previous studies indicating FTL has gastroprotective and antiulcerogenic effects not unlike the action of other
wellestablished gastroprotective drugs.
EVALUATION OF ANTIOXIDANT ROLE OF ORGANIC COMPOUNDS OF SELENIUM IN ANIMAL MODEL OF
ULCERATIVE COLITIS
LUIZ C. VIEIRA; MARIANA MENDONÇA; MOEMA LEITE TOURNIER; BARBARA PIACENTINI FEREIRA; LUCINÉIA
GAINSKI DANIELSKI; MONIQUE MICHELS; ANDRIELE VIEIRA; DRIELLY S. FLORENTINO; FABRÍCIA C.
PETRONILHO.
UNISUL, TUBARÃO - SC - BRASIL.
1 Clinical and Experimental Physiopathology Laboratory, Post-graduate Program in Health Sciences, Unisul, Tubarão,
SC, Brazil.
Introduction: Ulcerative colitis is a chronic inflammatory disease of the gastrointestinal tract whose etiology is
implicated in the altered state of the oxidant / antioxidant mechanism in the inflamed colon. As result of the
inflamed mucosal, evidences suggest that reactive oxygen species (ROS) are produced in excess leading to a
disturbance in the membrane stability and cell death by lipid peroxidation and protein damage. Studies show that
diphenyl diselenide (DD), an organic compound of selenium with ebselen (EB) can mimic antioxidant
enzymes.Objective: Thus, our goal is to study the effect of the organic selenium compounds on oxidative stress
parameters in induced ulcerative colitis by Dextran Sulfate Sodium (DSS) in rats. Method: Male Wistar rats were
induced to the ulcerative colitis model by DSS in the animal water container for 5 days. The rats were divided into 6
groups (n = 5): water + oil, Water + DD (10mg/kg p.o), water + EB (2mg/kg i.p), DSS + oil, DSS + DD (10mg/kg p.o),
DSS + EB (2mg/kg i.p). On the fifth day the animals were killed by decapitation and the colon was removed and
carried out analysis of oxidative damage in lipids by formation of thiobarbituric acid reactive substances and protein
carbonyls, besides antioxidant enzymes catalase activity and superoxide dismutase. Data were evaluated by ANOVA
and post hoc Tukey test with significance p <0.05. Results: We found that DSS increased oxidative damage in lipids
and proteins and that the compounds DD and EB were effective in reducing the damage. We also observed a
decrease in antioxidant enzyme activity with DSS. However, DD and EB showed an increased of these values, a
greater increase has been seen with DD. Conclusion: We conclude that organic selenium compounds studied in this
work show themselves an effective action against the damage caused by oxidative stress in an animal model of
ulcerative colitis.
Financial support: CNPq, UNISUL.
EVALUATION OF INFLAMMATION IN TYPE 2 DIABETES MELLITUS THROUGH THE INTRACELLULAR
SIGNALING PATHWAYS
1
2
3
NATHALIA TEIXEIRA PIETRANI ; KATHRYNA FONTANA RODRIGUES ; ADRIANA APARECIDA BOSCO ;
LIRLÂNDIA PIRES DE SOUSA4; KARINA BRAGA GOMES5.
1,2,4,5.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE - MG - BRASIL; 3.SANTA CASA DE
BELO HORIZONTE, BELO HORIZONTE - MG - BRASIL.
Introduction: The obesity is associated to type 2 diabetes mellitus (T2DM) and both are related to cellular insulin
resistance. Insulin resistance has been attributed to increased release of inflammatory cytokines by macrophages,
lymphocytes and adipocytes. Into the cell, the insulin receptor activates some downstream pathway that control
energy homeostasis, including PI3K/AKT and MAPKs that comprise p38 protein. The PI3K/AKT pathway is mainly
responsible for insulin action on glucose uptake and suppression of gluconeogenesis, whereas MAPK pathway
regulates gene expression and furthermore interacts with PI3K/AKT to control cell growth and differentiation. In
addition, these pathways are involved in expression of cytokines genes as IL-6 and TNF-a. The present study aims to
evaluate the intracellular signaling pathway of PI3K/AKT and MAPK (p38) in individuals with T2DM, comparing these
variables between patients with different body mass index (BMI) and a control group.
Methods and results: This study included 39 diagnosed T2DM patients and 32 control subjects without diabetes.
Patients and controls were classified according to BMI: BMI<25kg/m 2 - lean; 25≤BMI<30kg/m 2 – overweight; and
BMI>30kg/m2 - obese. A sample of peripheral blood was obtained from each subject. The proteins AKT and p38 were
analyzed by western blot, using anti P-AKT and anti P-p38 antibodies. Analysis involving just the subjects with T2DM
showed higher AKT levels in obese (0,952 ± 0,089) than lean (0,459±0,231) (p<0,001) and in obese than overweight
(0,596±0,325) (p=0,001). Higher p38 levels in obese (0,901±0,099) compared with patients with BMI below 25
kg/m2 (0,532±0,278) (p=0,002) and overweight (p=0,003) were also observed. The analysis only with control group
showed significance differences only for p38 between lean (0,496±0,285) and obese group (0,872±0,179) (p=0,015)
levels that rising with the increase weight. In the comparison between case and control groups was observed that only
the AKT protein was higher in obese diabetic (0,952 ± 0,089) than obese control group (0,339±0,214)
(p=0,005). Conclusion: Increasing of AKT protein levels is related to obesity in T2DM patients and p38 high levels
are associated with obesity, independently of the presence of T2DM.
Financial support: FAPEMIG and CAPES
FREQUENCY OF PERIPHERAL T AND B LYMPHOCYTES IN PATIENTS WITH HELICOBACTER PYLORIINFECTION
DALILA NUNES CYSNE1; SELMA SANTOS MALUF2; DANIEL MONTE FREIRE CAMELO3; IZABEL CRISTINA
4
5
6
PORTELA BOGÉA SERRA ; JOHNNY RAMOS DO NASCIMENTO ; BRUNO DE PAULO RIBEIRO ; ANA PAULA
7
8
SILVA DE AZEVEDO DOS SANTOS ; NATALINO SALGADO FILHO ; FLÁVIA RAQUEL FERNANDES DO
9
NASCIMENTO .
1,3,4,5,6,7,9.IMMUNOPHYSIOLOGY LABORATORY/FEDERAL UNIVERSITY OF MARANHÃO, SÃO LUÍS - MA BRASIL; 2,8.MASTER PROGRAM IN HEALTH SCIENCES/FEDERAL UNIVERSITY OF MARANHÃO, SÃO LUÍS MA - BRASIL.
Introduction: Helicobacter pylori infects more than 50% of the world’s population. Although most of infected
individuals remain asymptomatic, about 15-20% of H. pylori-positive individuals will develop peptic ulcer, gastric
carcinoma or mucosa-associated lymphoid tissue lymphoma. Recent studies have demonstrated that the infection by
this bacterium cause local damage in the gastric mucosa as well as influence the clinical course of other diseases, as
respiratory, vascular, autoimmune or other disorders. These extragastric manifestations can be justified by a systemic
cellular immune response evoked by this bacterial infection. Thus, this study aims to analyse the frequency
of CD3+ and
CD19+ peripheral
lymphocytes
from patients infected
with
this
bacterium.
Methods and Results: A total of 31 patients were involved in this study. 22 of them had dyspeptic symptoms,
underwent esophagogastroduodenoscopy and were positive for H. pylori infection by histopathology and/or urease
test. 9 patients without dyspeptic symptoms were considered as controls. Blood samples were collected of all the
individuals and were used to isolation of peripheral blood mononuclear cells (PBMC) by the standard density gradient
technique (Ficoll-Paque™ PLUS). Approximately 1 x 106 cells were stained with fluorochrome conjugated with antiCD3 e –CD19 antibodies (BD Biosciences, San Jose, CA, USA). The samples were read by flow
cytometry (BD FACSCalibur - BD Biosciences) and analyzed with FlowJo software (TreeStar) and GraphPad Prism
6.0. We find no difference (p > 0.05) in the frequency of CD3+ T cells (number of cells/mm3) between the infected
patients and controls: 2385.1 ± 1315 and 2806.8 ± 1715.7, respectively. No difference either was observed with
frequency of CD19+ B cells: 388.9 ± 299.8 and 488.1 ± 351.7. These results are corroborated for other studies and
indicate that the infection with H. pylori use different mechanisms for impair the immune response that is not a
quantitative change in the expression of the CD3 and CD19 proteins of circulating lymphocytes. Other researches are
necessary
to
elucidate
what
aspects
about
these
cells
this
bacterial
infection
affects.
Conclusion: The presence of H. pylori did not affect the frequency of peripheral T and B lymphocytes in the infected
individuals.
Financial support: CNPq, FAPEMA, UFMA.
HIGH FAT DIET IN A SHORT PERIOD OF TIME LEADS TO OBESITY AND INCREASED GENE EXPRESSION OF
TNF-Α AND IL-6 IN RENAL TISSUE
MARIA EUGENA LOPES NAVARRO1; DAMIANA TORTOLERO PIERINE2; FABIANE VALENTINI FRANCISQUETI3;
4
5
ANDRÉ FERREIRA NASCIMENTO ; RENATA DE AZEVEDO MELLO LUVIZOTTO ; ANA LÚCIA ANJOS
6
7
FERREIRA ; CAMILA RENATA CORREA .
1.NUTRITION STUDENT, INSTITUTE OF BIOSCIENCES, BOTUCATU - SP - BRASIL; 2,3,7.DEPARTMENT OF
PATHOLOGY, BOTUCATU MEDICAL SCHOOL, SÃO PAULO UNIVERSITY (UNESP), BOTUCATU - SP - BRASIL;
4,5,6.BOTUCATU MEDICAL SCHOOL, SAO PAULO UNIVERSITY (UNESP), BOTUCATU - SP - BRASIL.
Introduction: Obesity is defined as abnormal or excessive accumulation of body fat that may impair health; affects
individuals of all ages, sex and social class. The major comorbidity is renal injury. Several factors can trigger kidney
disease in obese people, among them the inflammation. Aim: The aim was to investigate the gene expression of
inflammatory cytokines in renal tissue of rats subjected to a high fat diet in a short period of time. Methods and
Results: We used 16 male Wistar rats randomly assigned to receive control diet (C, 5% fat) or high fat diet (HF, 29%
fat + sugar water in the drinking water, and 300 g/L) for six weeks. The nutritional parameters were evaluated: water
intake, diet and calories. Obesity was characterized by adiposity index (sum of the three fat deposits normalized by
body weight), the gene expression of inflammatory cytokines: tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6)
and protein monocyte chemoattractant (MCP-1) were made by PCR in real time. The results were presented as mean
± standard deviation and compared between groups made by Student's t test. The significance level for all analyzes
was 5%. The caloric intake between C and HF was similar: C=746±58 vs HF= 651±72, p=0.2772) The animals
showed a weight gain similar (C=32±9 vs HF=33±9, p= 0.481, in contrast), HF mice showed an index of adiposity 67%
higher compared to C (C=4.60 ±0.9 vs HF=7.70±1.5, p< 0.001)The gene expression of TNF-α (C=1.00±0.24 vs
HF=1.51±0.57, p=0.034) and IL-6 (C=1.00±0.30 vs HF=1.61±0.47 , p= 0.0081) was higher in group HF. The
expression of MCP-1 does not vary between groups (C=1.00±0.51 vs HF=0.84±0.37, p= 0.48). Conclusion: Based
on these data, we conclude that the diet led to obesity status, which was associated with increased in gene
expression of inflammatory cytokines in renal tissue.
IL-6 AND TNF-ALPHA LEVELS DURING EXERCISE PRACTICE IN ADOLESCENTS
MARIA REGINA MORETTO1; CILMERY SUEMI KUROKAWA2; CARLA CRISTIANE SILVA3; CRISTINA MARIA
TEIXEIRA FORTES4; TAMARA BERES LEDERE GOLDBERG5.
1,2,4,5.CLINICAL AND EXPERIMENTAL PEDIATRIC RESERCH CENTER, DEPARTMENT OF PEDIATRICS,
BOTUCATU SCHOOL OF MED, BOTUCATU - SP - BRASIL; 3.DEPARTAMENT OF PHISICAL EDUCATION OF
NORTH PARANÁ, MARINGÁ - PR - BRASIL.
Introduction: Interleukin-6 and TNF-alpha are produced during exercise by the skeletal muscle and have been
associated with glycogen balance during exercise and suppression of GH and IGF-I axis. Aim: The present study
measures the IL-6 and TNF-alpha levels in three groups of adolescents who practice three different modalities of
sports. Methods: The groups studied were divided by swimmers (n=14), tennis players (n=10), indoor soccer players
(n=19), and control group (n=14). The control group was composed of adolescents without any athletic activity. Each
subjective was categorized by bone age and genital maturation. The adolescents that practiced sports were under
exercise training during 30 minutes. The sample blood was collected before and after the exercise and IL-6 and TNFalpha were measured by ELISA. The data was analyzed with non-parametric test with 5% of
significance. Results: The IL-6 values of group control (median= 245 ) was lower than the athletes (swimming-IL6/median= 441); Tennis (IL-6/median=253); indoor soccer (IL-6/median=713). In the pre and post practice situation
showed increased values of IL-6 in indoor soccer and swimming. The TNF-alpha showed increased after activity in
swimmers players (p=0.02). Conclusion: The IL-6 and TNF-alpha is higher in athletes than non-athlete adolescents
and IL-6 and TNF-alpha levels differ between sportive modalities before and after their practice.
Supported by FAPESP- 2011/05991-0, 2007/07731-0, 2004/07007-1.
IMMUNE ABNORMALITIES IN AN ALLOIMUNIZATION FETUS WITH 29 WEEKS OF GESTATION
JULIANA ARAÚJO DE CARVALHO SCHETTINI1; THOMÁS VIRGÍLIO GOMES2; DIOGO CORDEIRO DE QUEIROZ
SOARES3; LEURIDAN CAVALCANTE TORRES4.
1,2,4.TRANSLATIONAL RESEARCH LABORATORY, INSTITUTO DE MEDICINA INTEGRAL PROF. FERNANDO
FIGUEIRA (IMIP), RECIFE - PE - BRASIL; 3.MEDICAL GENETICS UNIT, HOSPITAL DAS CLÍNICAS, FACULDADE
DE MEDICINA, UNIVERSIDADE DE SÃO PAULO (USP), SÃO PAULO - SP - BRASIL.
Introduction: The Rhesus (D) blood group is the second most common red blood cell antigen group involved in
hemolytic disease of the fetus and newborn (HDFN). D- positive fetal red blood cell can immunize D-negative women
to make anti-D that can cross the placenta causing fetal immune-mediated hemolysis and fetal hypoxia. This can be
an environmental factor aging on the fetal immune system determining the switch from the fetal to the adult form. The
aim of this study was perform an immunologic evaluation of RhD fetus of RhD and C alloimmunization
mother´s. Methods and Results: We performed the immunologic evaluation in fetus of a singleton pregnant in the
29th week of gestation complicated RhD and C alloimmunization, who undergone intrauterine fetal transfusion. Fetal
blood sample was obtained prior transfusion by ultrasound-guided cordocentesis. The immunophenotyping of T, B,
NK and NKT type II cells and proinflammatory cytokines (IL-6, IL-8, IL1-beta and TNF-alfa) was performed by flow
cytometry. The fetus presented low levels of T and TCD8+ cells. High relative values of TCD4+, normal relative values
of B and NK cells were found. B memory and NKT cells were absent. Increased of IL-8 in serum when compared to
his mother was observed. Conclusion: In our study we observed that the immune abnormalities observed may be
related to RhD and RhC antigen stimulation process and that NKT cells differentiation are possible related to thymus
development.
Financial support: CNPq – Grant Nº 473436/2011-6.
IMPACT OF CAFFEINE AND/OR ESTROGEN DEFICIENCY ON RANKL AND OPG EXPRESSION: A STUDY IN
RATS.
MARTA FERREIRA BASTOS; DIOGO JOSÉ BARRETO MENEZES; JOYCE PINHO BEZERRA; POLIANA MENDES
DUARTE.
GUARULHOS UNIVERSITY, GUARULHOS - SP - BRASIL.
Introduction: Estrogen deficiency plays a predominant role in the early and late phases of bone loss in
postmenopausal women and in the continuous phase of bone loss in aging men. Caffeine consumption has been
reported as a possible risk indicator for osteoporotic disorders, decreasing the bone density, and then increasing the
risk of fracture in clinical and animal studies. The molecular mechanisms involved in bone resorption have involved
two key molecules as regulators of osteoclastogenesis: the receptor activator of NF-КB ligand (RANKL) and
osteoprotegerin (OPG). Therefore, the objective of the present study was to evaluate the effects of caffeine
consumption and/or estrogen deficiency on the RANKL/OPG ratio in intact bone and in a surgically-created defect in
bone of rats.
Methods and Results: Rats were assigned into one of the groups: Control: non-ingestion of caffeine/sham-surgery
(n=15); Caffeine: ingestion of caffeine/ sham-surgery (n=15); OVX: non-ingestion of caffeine/ovariectomy (n=15);
Caffeine/OVX: ingestion of caffeine/ovariectomy. On the 57th day after the beginning of caffeine administration and 43
days after ovariectomy, a critical-sized defect was created in tibiae while the femurs remained intact. Rats were killed
8 days later. Femurs and tibiae were evaluated for the number of positive cells for RANKL and OPG by
immunohistochemistry. ANOVA test was used to demonstrate the significance of differences among groups. If
significance was detected, the pair-wise multiple comparisons Tukey test was used to assess differences between
groups. The femurs of the OVX (1.16 ± 0.14) and caffeine/OVX groups (1.51 ± 0.23) demonstrated higher ratios of
RANKL/OPG+ cells than the control group (0.86 ± 0.03, p<0.05). The defects of the caffeine (1.07 ± 0.20), OVX (1.23
± 0.19), and caffeine/OVX groups (1.50 ± 0.38) presented greater ratios of RANKL/OPG+ cells than that of the control
group (0.71 ± 0.23, p<0.05). In addition, the femurs and defects of the caffeine/OVX group presented the greatest
proportion of RANKL/OPG+ cells (p<0.05).
Conclusion: Caffeine consumption affected the RANKL/OPG ratio in bone defects while estrogen deficiency mainly
affected the RANKL/OPG ratio in intact bone of rats.
Financial Support: This study was supported by São Paulo State Research Foundation (FAPESP, São Paulo, São
Paulo, Brazil, 2009/09446-6).
IN SITU EVALUATION OF CD3+ CD4+ CELLS AND CD3+ CD8+ CELLS IN VENOUS ULCERS TREATED WITH
GLUCAN
SARAH DANTAS VIANA MEDEIROS1; FERNANDA GUEDES LUIZ2; LUANDA BÁRBARA FERREIRA CANÁRIO
3
4
5
SOUZA ; KARINA MENDES MELCHUNA ; KEYLA BORGES FERREIRA ROCHA ; GERALDO BARROSO
6
7
CAVALCANTI JÚNIOR ; TEREZA NEUMA SOUZA BRITO ; IRAMI ARAÚJO FILHO8; HUGO ALEXANDRE
9
10
OLIVEIRA ROCHA ; VALÉRIA SORAYA FARIAS SALES .
1,3,4,5,6,7,8,9,10.FEDERAL UNIVERSITY OF RIO GRANDE DO NORTE, NATAL - RN - BRASIL; 2.PASTEUR
INSTITUTE, SÃO PAULO - SP - BRASIL.
Introduction: Glucan is a polymer of glucose that has been reported to accelerate wound healing through the
activation of immune cells. In this context, T-lymphocytes produce cytokines and growth factors and function as
immunological effector cells. Venous ulcers are cutaneous wounds that result from chronic venous insufficiency. This
wounds are difficult to heal and responsible for 80-85% of ulcers that occur on the lower limbs. This study aimed to
detection and quantification of CD3+ CD4+ cells and CD3+ CD8+ cells in venous ulcers treated with
glucan. Methods: A nonrandomized clinical trial with intragroup comparisons over time was performed. Waterinsoluble glucan was isolated from the baker’s yeast Saccharomyces cerevisiae and was dispersed in cream with
crodabase CR2 to a final concentration of 3%; the cream was applied directly to the ulcer. A venous ulcer biopsy was
taken before therapy initiation (day 0) and after 30 days of glucan treatment for detection and quantification of
CD3+ CD4+ cells and CD3+ CD8+ cells by immunohistochemical evaluation with monoclonal anti-CD3+, anti-CD4+ and
anti-CD8+. The CD3+ CD4+ cells and CD3+ CD8+ cells were counted in 7 microscopic high power fields per specimen
with graticule (magnification 400X; 0.25mm wide with an area of 0.0625 mm 2). The number of the cells in each field
was counted and the mean of all fields was calculated. The differences between before therapy and after glucan
treatment were evaluated by the Wilcoxon test. Results: We followed 12 patients; within this study group, 1 patient
had 2 ulcers. Nine were women and three were men. The patients had an average of 59 +/- 10.5 years. The average
ulcer lifespan was 142.9 months (range of 8 to 264 months). Initially, the venous ulcers possessed an average area of
27.5 cm2 (range of 8.4 to 66.9 cm2) Histomorphometric analysis of CD3+ CD4+ cells before treatment showed a median
of 251.4 cells/mm 2 (minimum: 36.6 and maximum: 484.6) and CD3+ CD8+ cells showed a median of 115.4
2
+
+
cells/mm (minimum: 13.7 and maximum: 246.9). After 30 days of treatment the CD3 CD4 cells showed a median of
2
+
+
198.9 cells/mm (minimum: 80.0 and maximum: 562.3; p=0.0001) and CD3 CD8 cells showed a median of 97.2
2
cells/mm (minimum: 16.0 and maximum: 466.3; p=0.001). Conclusion: Glucan was able to promote the activation of
T-lymphocytes in the venous ulcers treated.
INCREASED CIRCULATING TNF-Α, TGF-Β1 AND IL-10 ARE ASSOCIATED WITH BOTH CHEMORESISTANCE
AND MULTIPLE METASTASIS SITES IN WOMEN BEARING CHEMORESISTANT BREAST CANCER.
CAROLINA PANIS1; ADRIANO MF ARANOME2; VANESSA J VICTORINO3; RUBENS CECCHINI4.
1,4.UEL, LONDRINA - PR - BRASIL; 2.UENP, BANDEIRANTES - PR - BRASIL; 3.USP, SÃO PAULO - SP - BRASIL.
Introduction: Breast cancer is the main malignant neoplasia among women worldwide. It is estimated that 30-50% of
patients undergoing chemotherapy develop chemoresistance. Several factors are enrolled in chemoresistance
acquisition; however, the clinical understanding of the cytokine profile of patients is still poorly described.
Methods: In this study we determined the circulating levels of cytokines in women carrying chemoresistant breast
tumors .This study was approved by Research and Ethics National Council. A total of 61 women diagnosed with
ductal infiltrative carcinoma of the breast were recruited. All women received a chemotherapic regimen based on 4
cycles of doxorubicin 60 mg/m2 and cyclophosphamide 600 mg/m2 each 21 days during 12 weeks, followed by
paclitaxel 175 mg/m2 each 21 days during 4 weeks. Women were categorized as responsive if they presented disease
remission or tumor size reduction, while were considered chemoresistant (CR) if presented either disease progression
or no reduction in tumor size after the complete chemotherapic cycle (16 weeks). Interleukin 10 (IL-10), transforming
growth factor beta 1 (TGF-β1) and tumor necrosis factor alpha (TNF-α) levels in plasma were measured by ELISA
kits. Data were analyzed by Mann-Whitney or Student t-test.
Results: A total of 32 women were diagnosed as bearing chemoresistant tumors. Increased levels of IL-10 (from
20.2±4.8 pg/mL in responsive to 47.1±13.5 pg/mL in CR, p=0.049), TGF-β1 (from 15.2±2.2 pg/mL in responsive to
28.7±5.7 pg/mL in CR, p=0.0250) and TNF-α (from 21.9±7. pg/mL in responsive to 24.19±5.8 pg/mL in CR, p=0.0414)
were found in CR when compared to the responsive cohort. TNF-α levels in CR patients correlated positively with both
IL-10 (r = 0.5069, p=0.043) and histological grade of tumors (r = 0.4506, p=0.0125), but negatively with TGF-β1 (r = 0.3865, p=0.0483). After disease recurrence, CR patients presenting high levels of IL-10 (96.3±31.6pg/mL) and TNF-α
(40.4±12.1pg/mL) remained alive when compared to the ones who died (15.1±1.8pg/mL to IL-10 and 11.8±3.1pg/mL
to TNF-α). Regarding the metastatic behavior of tumors, IL-10 levels were 9.34±4 pg/mL in lung metastasis, 14.6±3.8
pg/mL in bones and 99.9±29.2 pg/mL in multiple metastasis; TNF-α levels were 7.9±2.6 pg/mL in lung metastasis,
5.7±1.2 pg/mL in bones and 60.1±15.1 pg/mL in multiple metastasis and TGF-β1 levels were 21.4±5.9 pg/mL in lung
metastasis, 37.1±14.1 pg/mL in bones and 35.4±13.5 pg/mL in multiple metastasis.
Conclusions: These data suggest that TNF-α, IL-10 and TGF-β1 are enrolled in the acquisition of chemoresistance in
breast cancer and associate with the occurrence of multiple sites of metastasis. Further, sustained high levels of IL-10
and TNF-α are determinant to the survival of patients bearing chemoresistant breast cancer. Altogether these data
reveal a new perspective regarding the role of cytokines in breast tumor phenotype.
INFLAMATORY RESPONSE IN PATIENTS WITH STABLE ANGINA AFTER CORONARY STENTING: A
PROSPECTIVE STUDY
NATALIA MUSIKHINA; TATIANA PETELINA; NATALIA DEMENTYEVA; LYUDMILA GAPON; VADIM KUZNETSOV.
TYUMEN CARDIOLOGY CENTER, TYUMEN - RÚSSIA.
Aim: To assess the dynamics of lipid profile and inflammatory response in patients with stable angina (SA) after
coronary stenting.
Methods and Results: A total of 58 patients (mean age 59.3±9.0 years) with SA and significant coronary artery
stenosis >75% undergoing percutaneous coronary intervention (PCI) with drug-eluting stent placement were
examined. The parameters were evaluated at baseline and 3, 6 and 12 months after PCI. All patients received optimal
medical treatment that included statins and dual antiplatelet therapy. Lipid profile parameters, inflammatory markers:
high-sensitivity C-reactive protein (hs-CRP), tumor necrosis factor-alpha (TNF-alpha), homocysteine, interleukine 1 β,
6, 8, 16; CD40; soluble CD40 ligand (sCD40 L), matrix metallopeptidase (MMP-9), tissue inhibitor of metalloproteinase
(TIMP) -1; endothelial dysfunction markers (endothelin-1, nitrites) were measured.
High levels of atherogenic index, hs-CRP, TNF-alpha, MMP-9, CD40, sCD40L, homocysteine, endothelin-1 were
found initially as well as the following positive correlations were detected: between homocysteine and APO B/A-1, CD
40, MMP-9. After 3 months significantly elevated levels of homocysteine, MMP-9 and TIMP-1 and valid reduction of
TNF-alpha, CD40, sCD40L, endothelin-1 were revealed. In 6 month MMP-9, TNF-alpha, hs-CRP remained high,
however level of sCD40L was elevated. After 12 months of follow-up we detected reduction in atherogenic index, total
cholesterol, triglycerides, MMP-9, TNF-alpha, and significant increase in the concentration CD40. Endothelial
dysfunction was also observed: the level of endothelin-1 remained high.
Conclusions: The study demonstrated that inflammatory response in the vascular endothelium at 3 and 6 months
after coronary stenting indicates combined chronic inflammation caused by acute atherothrombosis and stent-induced
vascular injury. To reduce the risk of restenosis more careful monitoring of patients is required during follow-up period.
Tyumen Cardiology Center provided financial support for the research.
INFLAMMATORY BIOCHEMICAL PARAMETERS AND FUNCTIONAL PARAMETERS OF ARTERIAL WALL IN
HYPERTENSIVE PATIENTS WITH ABDOMINAL OBESITY
LYUDMILA GAPON1; TATIANA PETELINA2; NATALIA MUSIKHINA3; VADIM KUZNETSOV4; KSENIA AVDEEVA5.
1.TYUMEN CARDIOLOGY CENTER, TYUMEN - RÚSSIA; 2,3,4,5.TYUMEN CARDIOLOGY, TYUMEN - RÚSSIA.
The aim of the investigation to study s functional parameters of vessel wall, 24-h blood pressure (BP) profile,
inflammatory biochemical parameters and correlation between these and with metabolic disorders in patients with
arterial hypertension (AH) and abdominal obesity (AO).
Methods: 248 patients were divided into 2 groups. The main group (gr.1) included 168 subjects (mean age
44.36±1.52 years) with AH degree I-III and AO (mass body index (MBI) 32,4±4,1 kg/m 2 ) and the control group (2) - 80
subjects (mean age 42.27±0.85 years) without metabolic disorders.
The parameters of sphygmography by VASERA VS-1000 «FUCUDA»; 24-hour blood pressure monitoring using
АВМР–04 MEDITEX device; biochemical parameters: inflammatory markers – homocysteine, high-sensitivity Creactive protein (hs-CRP), TNF-alpha, fibrinogen and ceruloplasmin and lipid parameters, diene conjugates (DC),
malonic dialdehyde (MD), catataze and were estimated
Results: In gr.1 there was registered significant increase in sphygmography indices (cardio-ankle vascular index CAVI, pulse wave velocity - PVW and anklebrachial AB) index (p<0.01) and in BPM mean 24-h and daytime systolic
BP (SBP), time and square indices, in night time SBP and DBP variability (p<0.001). In biochemical parameters there
was detected significant increase in inflammatory markers - homocysteine (27.48±10.53 vs 20.35± 10.13 , p<0.05)
and hs-CRP level (3.56±2.64 vs 2.48±0.12, p=0.014) and in total cholesterol (5.66±0.9 vs5.07±0.8 p<0.05), lowdensity lipoprotein cholesterol (3.65±0.92 vs 3.31±0.93, p=0.034), triglyceride level (1.97±1.09 vs 1.76±1.04,
p=0.002), MD and DC level (46.33±2.12 vs 33,15±1.33, p<0.05); and decrease of catalase level (p=0.01) and high
density lipoprotein cholesterol (1.17±0.31 vs 1.49±0.44, p=0.001) compared to gr. 2. Besides in gr.1 positive
correlation between parameters of sphygmography with inflammatory and lipid markers parameters (CAVI and
homocysteine, triglyceride; PVW and fibrinogen; ABI and LDL cholesterol, MDA, hs-CRP, ceruloplasmin); with
MBI, 24-h blood BP given the age and sex of patients was found.
Conclusion: The study showed that the biochemical markers of inflammation, along with the parameters of lipid
profile may be pathogenetic predictors of vascular remodeling, high blood pressure and cardiovascular complications
in patients with AH and AO.
Tyumen Cardiology Center provided financial support for the research.
INFLUENCE OF VITAMIN C SUPPLEMENTATION ON HEALING INTESTINAL ANASTOMOSIS IN
UNDERNOURISHED RATS.
DRIELLY SILVA FLORENTINO; BÁRBARA PIACENTINI FERREIRA; EDUARDO ANDRÉ BRACCI WALCZEWSKI;
MOEMA LEITE TOURNIER; LUCINÉIA GAINSKI DANIELSKI; MONIQUE MICHELS; ANDRÉ ANTUNES LAURIANO;
MARIANA GASPAR MENDONÇA; LUIZ CARLOS VIEIRA; FABRICIA PETRONILHO.
UNIVERSIDADE DO SUL DE SANTA CATARINA, TUBARÃO - SC - BRASIL.
Laboratório de Fisiopatologia Clinica e Experimental, Programa de Pós-graduação em Ciências da Saúde, UNISUL,
Tubarão, SC, Brasil.
Introduction: Several are the risks and complications which a patient undergoing gastrointestinal surgery with
anastomosis of the digestive tract is susceptible. The occurrence of abnormality in the communication between the gut
and abdominal cavity or hollow viscus, known as gastrointestinal fistula, has been the most feared by surgeons.
Which have several implications on morbidity and mortality of the patient. One of the main factors involved in the
onset of fistula is the debilitated nutritional status of the patient, which further increases the risk of this complication,
due to changes in the immune response, affecting the healing process. In this sense, studies show the influence of
vitamin C (Vit C) in stimulating the proliferation of fibroblasts, which are important matters in collagen synthesis and
the cellular structure, responsible for healing and of sustaining the surgical wound. Methods: Thus, in this study, male
Wistar rats were divided into three groups: group I ( nourished rats + anastomosis + water), group II (undernourished
rats + anastomosis + water), group III (undernourished rats + anastomosis + Vit C). Rats were subjected to a period of
protein-calorie deprivation with intent to undernourished them for 26 days until the time of surgery. Four days before
surgery, they were supplemented with Vit C (100mg/kg/day orally diluted in water). The animals were subjected to
laparotomy with total section of the middle portion of small intestine and anastomosis in a single plane, with
interrupted sutures using a 5-0 polypropylene thread. The animals were sacrificed on day 7 after surgery, for survival
analysis, macroscopic and biochemistry at the anastomosis site. Results: Related with macroscopic analysis, we
observed an increase of adhesions in group II more than in group I. Which were reverted in group III, with Vit C. About
survival rates, we observed a decreased in group II more than in the group I. However, the group III did not reverse
such values. Group III obtained hydroxyproline levels similar to the control group (group I), reversing the low levels
found in group II. Conclusion: Our results demonstrate that treatment with VitC preoperative surgery increases
intestinal anastomosis healing in rats malnourished.
Financial support: CNPQ e UNISUL.
INTERLEUKIN-10 IS AN INDEPENDENT BIOMARKER OF SEVERE TRAUMATIC BRAIN INJURY PROGNOSIS
FLÁVIA MAHATMA SCHNEIDER SOARES1; NICOLE
SCHWARZBOLD3; ANDRÉ BAFICA4; ROGER WALZ5.
MENEZES
DE
SOUZA2;
MARCELO
LIBÓRIO
1,3.CENTRO DE NEUROCIÊNCIAS APLICADAS (CENAP), HOSPITAL UNIVERSITÁRIO (HU), UFSC,
FLORIANÓPOLIS - SC - BRASIL; 2,4.LABORATÓRIO DE IMUNOFARMACOLOGA E DOENÇAS INFECCIOSAS
(LIDI), UFSC, FLORIANÓPOLIS - SC - BRASIL; 5.DEPARTAMENTO DE CLÍNICA,CENTRO DE NEUROCIÊNCIAS
APLICADAS (CENAP), HOSPITAL UNIVERSITÁRIO (HU), UFSC, FLORIANÓPOLIS - SC - BRASIL.
Flávia Mahatma Schneider Soares1 MSc, Nicole Menezes de Souza2 MSc, Marcelo Libório Schwarzbold1 MSc, MD,
André Bafica2 MD, PhD, Roger Walz* 1,3 MD, PhD.
1-Centro de Neurociências Aplicadas (CeNAp), Hospital Universitário (HU), Universidade Federal de Santa Catarina
(UFSC), Florianópolis, Brazil.
2-Laboratório de Imunofarmacologa e Doenças Infecciosas (LIDI), UFSC, Florianópolis, Brazil.
3-Departamento de Clínica Médica, HU, UFSC, Florianópolis, Brasil.
Introduction: Cytokines have been shown to be involved in traumatic brain injury (TBI).
Objectives: We investigated the independent association between serum levels of IL-10 and TNF-α and hospital
mortality of patients with severe TBI.
Methods: Serum IL-10 and TNF-α levels were determined at a median time (IQ 25/75) of 10 (5/18) h after severe TBI
in 93 consecutive patients and in randomly-selected patients with mild (n=18) and moderate (n=16) TBI. In patients
with severe TBI additional blood samples were analyzed 30 (22/37) and 68 (55/78) h after TBI. Age, gender, CT
findings, admission GCS and pupillary reactions, associated trauma and hospital mortality were controlled.
Results: Elevated serum levels of IL-10, but not TNF-α, correlate significantly with GCS severity (Sperman’s
correlation, p <0.0001) and were found to be associated with hospital mortality in severe TBI patients. Elevated IL-10
amounts remained associated with mortality (p=0.01) in a subset of patients with isolated severe TBI (n=74). Multiple
logistic regression analysis showed that higher IL-10 levels (>90 pg/ml) at 10 or 30 h after TBI were respectively 6
(OR 6.2, CI 95% 1.2 – 25.1, p=0.03) and 5 (OR 5.4, CI 95% 1.2 – 25.1, p=0.03) times more associated with hospital
mortality than lower levels (<50 pg/ml) independently of age, admission GCS as well as pupils and associated trauma.
Conclusions: Serum IL-10 levels may be a useful marker for severe TBI prognosis.
Financial support: Work supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico
(CNPq), Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Programa de Apoio aos Núcleos
de Excelência (PRONEX) and Fundação de Apoio a Pesquisa do Estado de Santa Catarina (FAPESC), all of Brazil.
Bibliography: 1-Lingsma HF et al. Lancet Neu 9(5):543-54, 2010. 2- MRC CRASH Trial Collaborators,BMJ
336(7641):425-9, 2008. Epub 2008 Feb 12. Our work has over 26 articles in the bibliography but we have no
characters to post them.
LYCOPENE SUPPLEMENTATION DOWN-REGULATES TNF-ΑLPHA LEVEL IN KIDNEY OF HYPERCALORIC
DIET-FED RATS
DAMIANA TORTOLERO PIERINE1; FABIANE VALENTINI FRANCISQUETI2; MARIA EUGÊNIA LOPES NAVARRO3;
4
5
6
RENATA AZEVEDO MELO LUVIZOTTO ; ANDRE FERREIRA NASCIMENTO ; ANA LUCIA FERREIRA ; CAMILA
7
RENATA CORRÊA .
1,2,3,7.PATHOLOGY, BOTUCATU MEDICAL SCHOOL AT SAO PAULO STATE UNIVERSITY (UNESP),
BOTUCATU - SP - BRASIL; 4,5,6.INTERNAL MEDICINE, BOTUCATU MEDICAL SCHOOL AT SAO PAULO STATE
UNIVERSITY (UNESP), BOTUCATU - SP - BRASIL.
Recently, the association between obesity and renal disease has been accepted, however, the physiopathological
mechanism of this association is not completely understood. Inflammation in kidney may link obesity to the
pathogenesis of chronic kidney disease. Therefore, anti-inflammation and antioxidant interventions may be the
potential therapies to prevent and treat obesity-related renal diseases. Objective: To evaluate the effect of lycopene
supplementation on inflammation in Kidney of obese rats. Methods: Male Wistar rats were first fed control diet (C,
n=7) or hypercaloric diet (HD, n=14) for 6 wks. Afterwards, the HD rats were randomized into two groups: HD [placebo
(corn oil)] or HD+L [lycopene (10 mg/Kg BW/day in corn oil)] by gavage for 6 wks. Body weight (BW:g) and adiposity
index (AI:%) were used as obesity indicators. Kidney function was evaluated by glomerular filtration rate (GFR:
ml/24h) and kidney TNF-alpha (ρg/g) by immunoassay. Results: The AI was increased in the HD and HF+Lyco
groups (C: 5.8±1.8 < HD: 9.4±1.6 = HD+L: 9.7±1.7; p<0.001). Although there was no difference in BW (C: 507.5±82.3
= HD: 568.2±70.9 = HD+L: 564.2±60.1±60.1; p=0.235) and GFR (C: 1.3±0.7 = HD: 1.0±0.4 = HD+L: 1.3±0.4; p=0,561)
between the groups. TNF-alpha in kidney increased in HD group and lycopene supplementation restored the high
level to the levels of the C group (C: 227.8±6.6 = HD+L: 227.4±5.4 < HD: 238.7±7.4;
p=0.014). Conclusion: Lycopene decreases inflammatory responses in kidney indicating its potential protective
against obesity-related inflammation and chronic kidney disease.
Financial Support: CAPES
MORPHOFUNCTIONAL CHARACTERISTICS OF INDUCED IMMUNOSUPPRESSION AND POSSIBILITY OF ITS
CORRECTION
OLGA V. LEBEDINSKAYA1; ELENA A. LEBEDINSKAYA2; NELLY K. AKHMATOVA3; ANATOLIY P. GODOVALOV4.
1,2,4.ACAD. E.A. WAGNER PERM STATE MEDICAL ACADEMY, PERM - RÚSSIA; 3.I.I. METCHNIKOV
RESEARCH INSTITUTE FOR VACCINES AND SERUM RAMS, MOSCOW - RÚSSIA.
Introduction. Cytostatic drugs are widely used in clinical practice in the treatment of cancer, autoimmune diseases
and the transplantation. It is known that the use of chemotherapeutic drugs causes dramatic immune system
suppression. Moreover up to the present no standards have been developed for immune disorders caused by long
courses of cytostatics.
Methods. The aim of this study is to identify the morphofunctional characteristics of immunosuppression induced with
cyclophosphan and to evaluate the corrective therapy with immunomodulatory drugs of different origin (animal Stimforte, bacterial - Immunovac VP-4 multicomponent vaccine, floral - Fucoidan). Experiments were conducted on
540 SVA and Balb/c mice and 80 outbred male rats.
Results. We studied pharmacodynamic of cyclophosphan and its metabolites in serum, leukocyte composition of
peripheral blood (PB), subpopulation structure of mononuclear leukocytes (ML) of spleen with flow cytometry,
cytotoxic activity of splenocytes, level of cytokines emission, phagocytic ability of peripheral blood ML, chemical
luminescence of peritoneal macrophages and morphohistochemical changes in lymphoid organs of animals at
intraperitoneal introduction of different doses of cyclophosphan alone or with subsequent correction by
immunomodulatory drugs..The studies found that the introduction of cyclophosphan resulted in a significant decrease
in the absolute number of leukocytes in PB and spleen of mice and rats, changes in lymphocyte subpopulation
composition and decreased cytotoxic activity and production of cytokines, changes in the structure of lymphoid
organs. All the investigated immune modulators are capable to resolve changes caused by cyclophosphan.
Conclusion. The advantage of Immunovac VP-4 vaccine is a more rapid normalizing effect, possibly, due to a wide
range of pathogen-associated structures in its composition. The revealed fact may be important for the prevention of
infectious complications in cancer patients with immunosuppression induced with chemotherapeutic drugs.
This work was supported by RFBR grant №11-04-96037r_ural_a and administrative body of Perm Region.
NITRIC OXIDE AND SUPEROXIDE DISMUTASE IN SYSTEMIC LUPUS ERYTHEMATOSUS: ASSOCIATION WITH
DISEASE ACTIVITY?
1
2
3
THAÍS GUIMARÃES BARREIRA ; NATÁLIA FONSECA DO ROSÁRIO ; JOSÉ CARLOS CARRARO EDUARDO ;
CRISTIANE DA FONSECA HOTTZ4; ANALÚCIA RAMPAZZO XAVIER5; ANDREA ALICE DA SILVA6.
1,2.PROGRAMA DE PÓS-GRADUAÇÃO EM PATOLOGIA/UFF, NITERÓI - RJ - BRASIL; 3,5,6.FACULDADE DE
MEDICINA/UFF, NITERÓI - RJ - BRASIL; 4.HOSPITAL UNIVERSITÁRIO ANTÔNIO PEDRO/UFF, NITERÓI - RJ BRASIL.
Introduction: Systemic lupus erythematous (SLE) is an autoimmune/inflammatory disease presenting alternated
periods of activity and remission. It is well known that oxidative stress has important relationship in pathogenic
mechanisms of cellular lesion. In this context, this work aims to analyze oxidative stress markers, such as nitric oxide
(NO) and superoxide dismutase (SOD), in SLE patient’s serum samples.
Methods and Results: Cross-sectional study enrolled 39 patients with SLE: 20 with active disease (SLEDAI ≥6), 10
with inactive disease (SLEDAI <6), and 9 healthy control subjects. NOx production was quantified by VanadiumGriess reaction and SOD enzyme was quantified using a sandwich enzyme immunoassay. SLE patients were 46 ±
10.69 years-old and control subjects 42 ± 9.83 years-old. NOx quantification show no differences between groups.
SOD concentration was higher in SLE group (927.8 ± 433.7 pg/mL, p = 0.02) when compared to the control group
(605.1 ± 203.7 pg/mL). When compared SLE patients with active and inactive disease no statistical differences were
observed.
Conclusion: We concluded that the SOD enzyme may be useful such as marker of SLE, but longitudinal analyses are
necessary to confirm the correlation between oxidative stress markers and SLEDAI. Despite the preliminary results,
the increasing the case series is running.
Financial support: Proppi-UFF, FAPERJ.
NK AND B CELL DEFICIENCY IN A FAMILY WITH MUCOPOLYSACCHARIDOSES TYPE II
NATALLY L GAYAO1; TALITA RS PATRIOTA2; MARCELO S KERSTENETZKY3; CAROLINA D LINDOSO4; MARIA
5
6
7
8
DO CARMO BM DUARTE ; DIOGO CQ SOARES ; LESLIE D KULIKOWSKI ; JOSE FRANCISCO FRANCO ;
9
10
CHONG AE KIM ; LEURIDAN C TORRES .
1,2,3,4,5,10.TRANSLATIONAL RESEARCH LABORATORY, INSTITUTO DE MEDICINA INTEGRAL PROF.
FERNANDO FIGUEIRA (IMIP), RECIFE - PE - BRASIL; 6,7,8,9.MEDICAL GENETICS UNIT, HOSPITAL DAS
CLÍNICAS, FACULDADE DE MEDICINA, UNIVERSIDADE DE SÃO PAULO, SÃO PAULO - SP - BRASIL.
INTRODUCTION: Mucopolysaccharidoses (MPSs) are a group of inherited metabolic disorders characterized by
deficient activity of catabolic enzymes in the lysosomes and its consequent abnormal accumulation of deposits of
glycosaminoglycans. The lysosomal dysfunction caused by this irregular storage is responsible for the clinical
manifestations seen in MPSs. Once the lysosome is also important for normal functioning of the immune system,
playing a key role in the expression of cellular membrane receptors, the presentation of antigens, the secretion of
cytokines and phagocytosis, we presume that these processes may be impaired in patients with MPS. Additionally,
the presence of recurrent respiratory infections in these individuals may be a clinical clue of the immune dysregulation
in MPSs. Given the above, the aim of this investigation was to evaluate the main mechanisms involved in cellular
immune response in a non-consanguineous Brazilian family with 8 individuals affected by MPS type II. METHODS
AND RESULTS: Was performed the immunophenotyping of T, B and NK cells and of NKG2D ligand expression by
flow cytometry in 8 patients with MPS types II. Additionally were measured IgG, IgM and IgA total antibodies. The
results showed that this MPSs family has a quantitative deficiency of NK and B cells. Normal values TCD4+, TCD8+
and IgG, IgM and IgA antibodies was observed. CONCLUSION: These results suggest that this MPS family has a
primary immunodeficiency, which may explain the occurrence of recurrent respiratory infections and indicate a greater
predisposition to the development of cancer, becoming imperative periodic screening these patients for these
diseases.Financial support: São Paulo Research Foundation (FAPESP) – Grant Nº 2010/52694-8.
NOVEL GENETICS VARIATIONS IN GATA2 GENE FOUND IN PATIENTS WITH CLINICAL PHENOTYPES
DISTINCT AMONG THEM.
EDGAR BORGES DE OLIVEIRA-JUNIOR1; JACINTA BUSTAMANTE2; GUILLAIME VOGT3; MARJORIE HUBEAU4;
ANTONIO CONDINO-NETO5; JEAN LAURENT CASANOVA6.
1,5.UNIVERSITY OF SÃO PAULO, SAO PAULO - SP - BRASIL; 2.PARIS DESCARTES UNIVERSITY, PARIS FRANÇA; 3,4,6.THE ROCKEFELLER UNIVERSITY, NEW YORK - ESTADOS UNIDOS.
The protein GATA-2 (GATA binding protein 2) is a transcription factor belonging to the GATA family (GATA1-6)
containing fields "zinc finder" promoting DNA binding regulation of gene transcription. These candidates have
emerged as regulators of gene expression in hematopoietic cells. GATA-2, whose GATA2 gene encoding this protein
is located on chromosome 3, it is widely considered a central regulator of "Hematopoietic Stem Cells" (HSCs) and
"Hematopoietic Progenitor Cells" (HPCs). Recently, mutations in the GATA2 gene has been associated with different
clinical phenotypes, such as myelodysplastic syndrome (OMIM # 614286), Chronic Myeloid Leukemia and Acute
(OMIM # 608232 and # 601626), Emberger syndrome (OMIM # 614038), monocytopenia associated with
mycobacterial infections, MonoMAC (OMIM # 614172). Knowing this, we selected 23 patients with clinical history of
myelodysplasia (chronic, acute) or leukopenia (monocytopenia, neutropenia), and who also had recurrent infections or
considered unusual, such as mycobacteria or virus. Through direct sequencing of the promoter and coding regions (7
exons) in GATA2 gene using genomic DNA. We found a new heterozygous variation not described in the literature:
Patient 1 (P1 - p.D367fs15X), female sex, severe and presents monocytopenia Mycobacterium avium-intracellular
complex (MAC) in culture from bone marrow. In addition, two patients were found carrying two different heterozygous
variations also not described in the literature by Exome sequencing: Patient 2 (P2) out of 93 in patients group
associated with TB (Tuberculosis), and another patient (P3) among 105 in the group associated with HSE (Herpes
Simplex Encephalitis). The variations was p.Y213D and p.G28S respectively. Differences found had a significant
impact on protein expression (Western blot) and activity functional (Luciferase Assay - site directed mutation),
suggesting that the new variation p.D367fs15X may impact the clinical phenotype of P1. Furthermore, in a preliminary
tests for detection by FACS, no differentiation of monocyte-derived dendritic cells in P1 and also at one of her sister,
that also has the same variation in GATA2 gene. New trials are being conducted to complete and determine the
impact of these new genetics heterozygous variations found in GATA2 gene.
PLASMA CYTOKINES PROFILE IN INFLAMMATORY BOWEL DISEASE DOES NOT CORRELATE WITH THE
DEGREE OF INTESTINAL LESION
1
2
3
RENAN GARCIA GOMES ; CARLOS ALEXANDRE DE BRITO ; VALÉRIA MARTINELLI ; EDUARDO ANTONIO
DONADI4; NORMA LUCENA-SILVA5.
1,5.AGGEU MAGALHÃES RESEARCH CENTER/FIOCRUZ, RECIFE - PE - BRASIL; 2,3.HOSPITAL DAS CLÍNICAS
DE PERNAMBUCO, RECIFE - PE - BRASIL; 4.FACULTY OF MEDICINE OF THE UNIVERSITY OF SÃO PAULO,
SÃO PAULO - SP - BRASIL.
Introduction: Inflammatory Bowel Disease (IBD) encompasses a group of inflammatory disorders of the
gastrointestinal tract including Ulcerative Colitis (UC) and Crohn's Disease (CD). Several factors have been linked to
the pathogenesis of IBD, including environmental factors, immunology and genetic background. One of the immune
mechanisms proposed referrers to the B cell hyperresponsiveness and auto-reactivity of T cells induced, in part, by
the response against infectious agents, food antigens or others; leading to polarization of the cellular immune
response, in UC for Th2 and in DC for Th1 response.
Methods and Results: We evaluated the level of cytokines expression in plasma of 98 patients in the acute phase of
the IBD using Th1/Th2 Bead Array technique (CBA, Beckton Dickinson). The median age of patients was 44 years,
67% of them were women. 27 patients were diagnosed with DC and 71 with UC. They were separated into two groups
based on the degree of intestinal inflammation: patients with mild or no inflammation (UC:21; DC:06) and patients with
moderate or severe inflammation (UC:50; DC:21). There were no differences of cytokines expression in plasma
between the inactive-mild and moderate-severe groups. Interleukin-5 was high expressed in patients with UC
compared with DC, irrespective of the severity of the inflammation.
Conclusion: High expression of IL-5 characterizes the Th2-immune response in UC patients. Further studies are
necessary for evaluation of the local immune response, correlation with the degree of inflammation, and comparison
with the systemic immune response in either acute or compensate clinic disease forms.
PRO-INFLAMMATOY CYTOKINES IN SERUM AND PLACENTA FROM WOMEN WITH HYPERGLYCEMIA
SIMONE CORREA-SILVA1; ALINE PAIXÃO ALENCAR2; JUSCIELE BROGIN MORELI3; LESSANDRA DE ROSA4;
5
6
7
8
DEBORA DAMASCENO ; MARILZA RUDGE ; ESTELA BEVILACQUA ; IRACEMA CALDERON .
1,3,4,5,6,8.UNESP, BOTUCATU - SP - BRASIL; 2,7.USP, SÃO PAULO - SP - BRASIL.
Introduction: Maternal hyperglycemia may adversely affect feto-maternal interaction. Several reports have
demonstrated that in diabetes pro-inflammatory state is constant. Cytokines have a key role in this condition. This
study was conducted to compare inflammatory cytokine levels (IL-6 and MCP-1) and iNOS in serum and placenta
from pregnant women with hyperglycemia. Methods and results: This cross-sectional study included 10 serum and
placental fragments (fetal side). Based on 75g-GTT (Glucose Tolerance Test) and glucose profile analysis (GP)
performed at 24-28 weeks of gestation, subjects were allocated into two groups: Nondiabetic (ND) and Hyperglycemic
[Overt diabetes, Gestational DM and Mild Hyperglycemia]. We analyzed placental IL-6 and MCP-1 levels by ELISA.
Differences between groups were analyzed using Student t test and Mann-Whitney test (p<0.05). Immunolocalization
and immunoquantification of iNOS and MCP-1 in placental fragments of these mothers were assessed by
immunohistochemistry. Conclusion: In serum and placenta IL-6 and MCP-1 levels are up regulated. In placenta we
observed higher levels of IL-6, MCP-1 and iNOS. These results suggest an increased inflammatory profile in the
maternal-fetal interface of pregnant women with any degree of hyperglycemia, which may be related to the placental
angiogenic disorders observed in these pregnancies. In conclusion, glycemic disturbance may affect both placental
and blood compartments.
Financial Support: CAPES and CNPq
PROBLEM IN LABORATORY DIAGNOSIS OF SYPHILIS
1
2
3
JULIANO LACAVA PEREIRA ; GABRIEL BARACY KLAFKE ; ANDREA VON GROLL ; ALEXANDRE NABAES
FERREIRA4.
1,2,3.FEDERAL UNIVERSITY OF RIO GRANDE, RIO GRANDE - RS - BRASIL; 4.LABORATORY SARETTA,
CAÇAPAVA DO SUL - RS - BRASIL.
1
Federal University of Rio Grande
2
Laboratory Saretta
Introduction: Syphilis is a sexual disease transmitted that has challenged public health strategies in the world.
Laboratory diagnosis of this infection is submited to a serie of problems due the lack of technical professionals in the
health field as laboratory methods. The aim of this study was to evaluate the professional qualifications in the fields of
immunology, diagnostic services related to syphilis in the state of Rio Grande do Sul – Brazil. Methods and
results: It was performed a technical questionnaire on laboratory diagnosis of syphilis using the software
SurveyMonkey®. The study evaluated 17 clinical laboratories and the results were expressed as a percentage. The
study showed that 94.1% of the laboratories use the VDRL (Veneral Disease Research Laboratory) as a method of
choice for serological screening of the disease, and 23.5% of the individuals did not make the dilutions of sera
recommended by manufacturers to avoid the prozone effect. In relation to the release of the reports of testing
reagents dilutions equal to or less than the title 1:8, it was observed that 64.7% release the reports without using an
alternative methodology for confirmation, in addition, 47.1% answered they that release the reports in a quantitative
manner and not using semi-quantitative VDRL. Conclusion: It is concluded that the lack of technical information in
the laboratories may refer to errors in the diagnosis of syphilis, is due to the a prozone or generate false positives.
PROFILE OF PROINFLAMMATORY AND ANTI-INFLAMATORY CYTOKINES AND MITOCHONDRIAL
PRODUCTS IN FULMINANT HEPATIC FAILURE
1
2
3
JULIANA GIL MELGAÇO ; YASMINE RANGEL VIEIRA ; LEONARDO ASSAF PINHEIRO ; LIA LAURA LEWISXIMENEZ4; FREDERICO SORIANI5; GUSTAVO BAPTISTA MENEZES6; MARCELO ALVES PINTO7; CLÁUDIA
8
LAMARCA VITRAL .
1,2,4,7.FUNDAÇÃO OSWALDO CRUZ, RIO DE JANEIRO - RJ - BRASIL; 3,8.UNIVERSIDADE FEDERAL
FLUMINENSE, NITERÓI - RJ - BRASIL; 5,6.UNIVERSIDADE FEDERAL DE MINAS GERAIS, BELO HORIZONTE MG - BRASIL.
Introduction: The fulminant hepatic failure (FHF) corresponded to 1% of total hepatitis, unique treatment is hepatic
transplantation. In this fulminant hepatic failure, the inflammation grade is extremely increased than acute infection,
most deaths before transplantation is sepsis, wherein the acute inflammatory liver tissue ruptures and releasing
endotoxins into circulation. Methods and Results:To study immunological mechanisms of fulminant hepatic failure
(FHF) derived from extensive liver lesions, 10 patients with FHF induced by viral hepatitis, 10 patients with FHF
induced by other causes (auto immune, acetaminophen, steroids), 10 patients with viral hepatitis acute, 10 patients
with other causes to acute hepatitis and 10 health controls (HC) were investigated by observance plasmatic cytokine
levels (IL6, IL8, IL10, TNF-alpha, IFN-gamma) and mitochondrial DNA (mitDNA) damage. To evaluate the cytokine
profile plasma quantitative immunoassay (ELISA) was performed. And to evaluate the rate of mitDNA in the
bloodstream molecular assays were performed using the RT-PCR assay. The results showed an increase NADH
dehydrogenase (2891.75 ng/100µL serum) and cytochrome C (1853.68 ng/100µL serum) in FHF comparing a HC
(0,018 ng/100µL serum and 0,025 ng/100µL serum, respectively) (p<0,05). The same was observed for acute patients
(75.70 Cyt C, 98.05 NADH) (p<0,05) when compare with fulminant patients. Then, in the cytokines plasmatic analysis,
there was an increase in plasmatic IL8, IL6 and TNF alpha in FHF with other causes, indicate neutrophil migration
(IL8, IL6) and active inflammatory process (TNF-alpha) (p<0,05). Levels of IL10 were elevated in patients with FHF
with other causes, suggesting a possible rescue attempt of the immune system against inflammatory process currents
(p<0,05). There was an increase in plasmatic IFN-gamma in patients with FHF to viral hepatitis, the levels were 27.28
fold in comparison with detected with health controls. Conclusion: These findings indicate already the inflammatory
process inducing FHF, confirmed by the high expression of cytokines circulation systemic in this cases. This work
provides strong support for the hypothesis that mitochondrial dysfunction and DNA damage are critical events in the
mechanism of cell necrosis after fulminant hepatitis and can be used in future diagnosis in fulminant cases.
PROGNOSTIC SIGNIFICANCE OF INFLAMMATORY MARKERS IN PATIENTS WITH CORONARY ARTERY
DISEASE AND DIABETES MELLITUS: A FOLLOW-UP STUDY
TATIANA PETELINA; NATALIA MUSIKHINA; NATALIA DEMENTYEVA; LYUDMILA GAPON; VADIM KUZNETSOV.
TYUMEN CARDIOLOGY CENTER, TYUMEN - RÚSSIA.
Purpose: To study prognostic significance of serum inflammatory parameters in patients with coronary artery disease
(CAD) and type 2 diabetes mellitus (DM) receiving standard medical therapy at prospective observation.
Design and methods: 68 CAD patients with stable angina aged 60.3±9.8 years were studied. Patients were divided
into two groups: group I - patients with CAD (n=29), group II - patients with CAD and type 2 diabetes mellitus (n=39).
All patients were with nonsignificant coronary stenosis (< 75%). Serum laboratory studies were performed at baseline
and after a follow-up of 12±2.4 months. Patients received standard medical therapy - ACE inhibitors, beta blockers,
antiplatelet therapy, statins. In group II patients also received antihyperglycemic therapy.
Results: At baseline the patients of group II had significantly elevated levels of atherogenic lipids (total cholesterol,
low-density lipoproteins (LDL cholesterol), Apo - B) and inflammatory markers (hs CRP, homocysteine, interleukin-1ß)
compared to group I. Group I had higher levels of soluble CD40L. In group II a greater number of significant
correlations were found between atherogenic lipid fractions (lipoprotein(a), LDL cholesterol ), inflammatory markers
(hs CRP, homocysteine, interleukins, TNF-alpha), endothelial dysfunction markers (endothelin-1), thrombogenic
factors (sCD40L) and glycohemoglobin. Follow-up demonstrated no significant positive changes in lipid profile and
persistence of prolonged inflammatory response indicated by interleukin-1ß, hs-CRP, homocysteine, matrix
metalloproteinases (MMP-9) and endothelial dysfunction in both groups. It was shown that with an increase of 1
μmol/L in homocysteine level, the risk of developing diabetes in patients with stable angina increased by 20%.
Conclusion: Our results showed that in patients with type 2 diabetes was a significant increase in markers of
inflammation, which may indicate a higher risk of coronary events, even in the absence of significant coronary
stenosis. The level of homocysteine was a predictor of the development of diabetes in patients with stable angina
pectoris.
Tyumen Cardiology Center provided financial support for the research.
PROSPECTIVE STUDY TO DETERMINE FACTORS CONTRIBUTING TO DISEASE EXPRESSION IN
CUTANEOUS LEISHMANIASIS PATIENTS
1
2
RUBIA SUELY SANTANA COSTA ; GIOVANA BERGHEME FRANCISCON ; LUIZ HENRIQUE SANTOS
GUIMARÃES3; SARA TIMÓTEO PASSOS4; PHILLIP SCOTT5; LUCAS PEDREIRA DE CARVALHO6; EDGAR
7
MARCELINO CARVALHO .
1,2,3,4,6,7.UFBA, SALVADOR - BA - BRASIL; 5.UNIVERSITY OF PHILADELPHIA, PHILADELPHIA - ESTADOS
UNIDOS.
​(1) Immunology Service, Federal University of Bahia, (2) University of Philadelphia, (3) National Institute of
Science and Technology-Tropical Diseases (INCT-DT), (4) Institute of Health Sciences (ICS), Federal University of
Bahia
Introduction. Cutaneous leishmaniasis (CL) due to L. braziliensis is an inflammatory parasitic disease characterized
by the development of ulcerated lesions in the skin. Mononuclear phagocytes migrate to bite site and are one of the
first cell types interacting with the parasite and secreting inflammatory mediators, such as TNF and CXCL-9. Three
subsets of circulating monocytes have been described based on the expression of CD14 and CD16: classical (CD16 CD14+), intermediate (CD14+CD16+) and (CD14+CD16+) monocytes non-classical. The study of immune response in
the early stage of diseases may reveal factors contributing to disease expression. The aim of this study was to
establish a prospective cohort study with CL patients in the very initial phase of the disease (before ulcer appearance)
to determine factors involved in ulcer development.
Methods and results. Peripheral blood mononuclear cells (PBMC) were obtained from healthy controls and patients
with early CL (ECL) (before ulcer appearance). Frequency of monocyte subsets where determined ex vivo by flow
cytometry, and levels of cytokines and chemokines in response to Leishmania antigen were determined by ELISA on
days 0, 15 and 30 post-treatment. It was observed that the frequency of the intermediate and non-classical monocytes
were increased in ECL individuals and the frequency of non-classical monocytes reduced 30 days after treatment with
pentavalent antimonial. Levels of TNF, CXCL9 and CXCL10 were higher in ECL than in healthy controls at day 0, and
levels of CXCL9 from ECL significantly decreased on day 15. It was also observed on day 30 after treatment a
significant decrease in the levels of TNF in supernatants from PBMCs.
Conclusion. These results suggest that non-classical monocytes, TNF and CXCL9 might contribute to ulcer
development, thus, participating in the pathogenesis of CL.
Financial support: CNPq, CAPES and NIH (AI030639 and AI088650-TMRC-ICIDR).
SERUM LEVELS OF TH17 ASSOCIATED CYTOKINES IN DYSLIPIDEMIC PATIENTS TREATED WITH
SIMVASTATIN
1
2
3
MARIANA DE MENEZES PEREIRA ; TACIANA PEREIRA SANT’ANA SANTOS ; ROQUE ARAS JÚNIOR ; AJAX
MERCÊS ATTA4.
1,2.PROGRAMA DE PÓS-GRADUAÇÃO EM IMUNOLOGIA DA UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR
- BA - BRASIL; 3.SERVIÇO DE CARDIOLOGIA, HOSPITAL ANA NERY, FACULDADE DE MEDICINA,
UNIVERSIDADE FEDERAL DA BAHIA, SALVADOR - BA - BRASIL; 4.DEPARTAMENTO DE ANÁLISES CLÍNICAS
E TOXICOLÓGICAS, FACULDADE DE FARMÁCIA, UNIVERSIDADE FEDERAL DA BA, SALVADOR - BA BRASIL.
Introduction: Atherosclerosis is the main cause of cardiovascular diseases (CVD) associated with dyslipidemia
worldwide. CVD immunopathogenesis has the involvement of macrophage, Th1 cells and T regulatory cells. However,
the involvement of Th17 cells in CVD pathogenesis is unclear. This work investigated the serum levels of some Th17
associated cytokines in dyslipidemic Brazilian patients treated with simvastatin, a medication with lowering lipid
properties and anti-inflammatory activity. Methods and results: Were included in this study 50 dyslipidemic patients
(31 men and 19 women) with a previous MI history (GI), 50 dyslipidemic patients (23 men and 27 women) without a
previous MI (GII) and 20 healthy individuals (10 men and 10 women) (GIII). Serum levels of IL-6, TGF-β, IL-17A, IL22, IL-8 and GM-CSF were determined by capture ELISA. Serum lipid profile was determined by biochemical analysis,
whereas apolipoprotein A (apoA1) and apolipoprotein B (apoB100) levels were determined by nephelometry. The
median levels of these serum components in the groups were analyzed by the Kruskal-Wallis test followed by the
multiple comparison posttest of Dunn. The levels of non-HDL-cholesterol differed in the groups (GI= 121.5 mg/dL, GII
= 138.5 mg/dL, GIII = 116.5 mg/dL; P = 0.0192), whereas their apoB/apoA ratios were similar (GI = 0.63, GII = 0.61
and GIII = 0.54; P > 0.05). Overall, simvastatin treatment caused important decrease in the serum levels of IL-6, IL17A, IL-22, IL-8 and GM-CSF, which were very low in most dyslipidemic patients. Patient and control groups differed
in their median levels of TGF-β (GI = 197.3 pg/mL, GII = 196.3 pg/mL and GIII = 121.2 pg/mL, P = 0.0053) and IL-8
(GI = 10.7 pg/mL, GII = 9.2 pg/mL and GIII = 17.2 pg/mL, P = 0.0081). No difference was observed in the median
levels of IL-17-A (GI = 103.4 pg/mL, GII = 70.1 pg/mL and GIII = 95.7 pg/mL), IL-22 (GI = 14.4 pg/mL, GII = 16.5
pg/mL and GIII = 9.5 pg/mL) and GM-CSF (GI = 11.3 pg/mL, GII = 12.3 pg/mL and GIII = 10.3 pg/mL). However, a
strong correlation was observed between the apoB/apoA ratio and GM-CSF levels in the dyslipidemic patients without
a previous MI (r = 0.61, P = 0.0484). Dyslipidemic patients treated with simvastatin present unaltered or very low
serum levels of Th17 associated cytokines, which are not influenced by a previous myocardial infarction. Nonetheless,
the use of simvastatin increases the serum levels of TGF-β in these individuals. Financial support CNPQ CAPES
SOLUBLE MICA: A POSSIBLE INVOLVEMENT IN THE PATHOGENESIS OF ENDOMETRIOSIS?
MARIA LUCIA CARNEVALE MARIN1; PATRICK BELLELIS2; SERGIO PODGAEC3; JORGE KALIL4; MAURICIO
SIMÕES ABRÃO5.
1.HEART INST. (INCOR), HISTOCOMPATIBILITY AND CELLULAR IMMUNOLOGY LAB.- LI1M9, SCHOOL OF
MEDICINE, SÃO PAULO - SP - BRASIL; 2,3,5.DEPARTMENT OF OBSTETRICS AND GYNECOLOGY SCHOOL OF
MEDICINE, SAO PAULO UNIVERSITY, SÃO PAULO - SP - BRASIL; 4.HEART INST. CLIN. IMMUNOL ALLERGY
DIV, LIM-19,SCHOOL MED., UNIV.SAO PAULO; IMMUNOLOGY INVEST. INST., SÃO PAULO - SP - BRASIL.
Introduction: Endometriosis is a chronic inflammatory disease, characterized by implantation and growth of
endometrial tissue outside the uterine cavity. It is associated with pelvic pain and infertility, affecting 10% of women in
reproductive age. Alterations in the immune system play an important role in the initiation and progression of the
disease. Reduced activity of cytotoxic T and NK cells has been observed and associated to immunological factors.
MICA (Major Histocompatibility Class I-related chain A), a stress inducible molecule present in gastrointestinal
epithelium, endothelial cells and fibroblasts, has the NKG2D as its activating receptor, which is present in CD8+ ab T
cells, gdcells and NK cells. Tumor growth has been associated with the presence of soluble MICA (sMICA) resulting in
decreased NKG2 expression and consequent blocking of cell activity. In endometriosis, sMICA has not been reported
yet. The aim of this study is to evaluate the presence of sMICA in women with endometriosis and compare it with
healthy controls (women without the disease).
Methods and Results: Seventy five women with endometriosis confirmed by videolaparoscopy and histology of
biopsies and 19 women undergoing laparoscopy for tubal ligation, without endomentriosis (control group) were
enrolled in this study. Serum and peritoneal fluid levels of sMICA were determined by a commercial ELISA kit
(R&DSystems) (limit of detection= 31pg/mL). Our results showed higher sMICA levels in the peritoneal fluid
(median=82pg/mL, 31-695) when compared to its serum levels (median=31pg/mL, 31-148) in endometriosis patients
(p < 0.0001), especially in severe stages of the disease. Soluble MICA levels in peritoneal fluid of controls were also
higher when compared to its serum levels (p= 0.0157). The comparison between sMICA serum levels between
patients and controls showed increased levels in women with endometriosis (p= 0.0395). The same was observed for
sMICA in peritoneal fluid (p= 0.0230).
Conclusion: Our preliminary results indicate increased levels of sMICA in endometriosis patients, particularly in the
peritoneal fluid. This could be due to the presence of lesions and consequent inflammatory environment, which
possibly contributes to deficient clearance of endometriotic cells, by the reduction of the cytotoxic activity of NK and
CD8+ cells, thus facilitating the implantation and growth of endometrial tissue
Financial support: FAPESP
SPECIFIC LABEL-FREE AND REAL-TIME DETECTION OF OXIDIZED LOW DENSITY LIPOPROTEIN (OX-LDL)
BY AN IMMUNOSENSOR WITH THREE MONOCLONAL ANTIBODIES
GUSTAVO CABRAL DE MIRANDA1; MAGNUS AKE GIDLUND2; Mª GORETI FERREIRA SALES3.
1,2.DEPARTMENT OF IMMUNOLOGY AT THE INSTITUTE OF BIOMEDICAL SCIENCES IV, UNIVERSITY OF SAO
PAULO (USP), SAO PAULO - SP - BRASIL; 3.BIOMARK/ISEP, SCHOOL OF ENGINEERING, POLYTECHNIC
INSTITUTE OF PORTO, PORTUGAL, PORTO - PORTUGAL.
Increased plasma level of oxLDL, the oxidized fraction of Low Density Lipoprotein (LDL), is associated with
atherosclerosis, an inflammatory disease, and the subsequent setup of severe cardiovascular diseases that are today
a major cause of death in modern countries. Thus it is important to find a reliable and fast assay to determinate oxLDL
in serum.
A new immunosensor employing three monoclonal antibodies (mAb) against oxLDL is proposed in this work as a
quick and reliable way to monitor oxLDL. oxLDL was first employed to produce anti-oxLDL monoclonal antibodies by
hybridoma cells previously obtained. The immunosensor was set-up by self-assembling cysteamine (Cyst) on the gold
(Au) layer (4 mm diameter) of a disposable screen-printed electrode. Three mAb were let react with NHydroxysuccinimide (NHS) and ethyl(dimethylaminopropyl) carbodiimide (EDAC), and subsequently incubated in the
Au/Cys. Albumin from bovine serum (BSA) was immobilized further to ensure that non-specific molecules could not
bind to the electrode surface. All steps were followed by various characterization techniques such as electrochemical
impedance spectroscopy (EIS) and square wave voltammetry (SWV).
The analytical operation of the immunosensor was obtained by incubating the sensing layer of the device in oxLDL for
15 minutes, prior to EIS and SWV. This was done by using standard oxLDL solutions prepared in fetal calf serum, in
order to simulate patient’s plasma with circulating oxLDL, and specific and sensitive response was still observed. The
device was successfully applied to determine oxLDL fraction in real serum, without prior dilution or necessary
chemical treatment.
The use of multiple monoclonal antibodies on a biosensing platform seemed to be a very attractive approach to
produce a specific response towards a complex multi-analyte target, correlating well the level of oxLDL within
atherosclerosis disease, in a simple, fast and cheap way.
T CELL ACTIVATION AND CYTOKINE PROFILE IN IMMUNE RECONSTITUTION INFLAMMATORY SYNDROME
IN PATIENTS TREATED FOR HIV AND TUBERCULOSIS IN RIO DE JANEIRO
TATIANA SILVA1; CARMEM GIACOIA-GRIPP2; CAROLINA SCHMALTZ3; FLAVIA SANTANNA4; VALERIA ROLLA5;
MARIZA MORGADO6.
1.FIOCRUZ- IOC, RIO DE JANEIRO - RJ - BRASIL; 2,6.FIOCRUZ-IOC, RIO DE JANEIRO - RJ - BRASIL;
3,4,5.FIOCRUZ-IPEC, RIO DE JANEIRO - RJ - BRASIL.
Introduction: Mycobacterium tuberculosis (Mtb) and HIV are two intracellular pathogens that successfully manipulate
the immune system in order to generate persistent infections. Both infections cause immune activation and, under
HAART, events like immune reconstitution inflammatory syndrome (IRIS) may occurs. The mechanisms associated
with the development of IRIS are not fully understood. Our objective is to evaluate if T cell activation and cytokine
profiles could be predictive for IRIS.
Methods and Results: A total of 54 patients with tuberculosis (TB) and HIV diagnosis were followed-up at the
Evandro Chagas Clinical Research Institute (FIOCRUZ/RJ) and evaluated at baseline (TB diagnosis and treatment
before HAART) and after HAART initiation (D30, 60, 90, 120, 150 and 180). TCD4+ cells counts, viral load, plasma
cytokines (IL-2, IL-4, IL-6, IL-10, TNF-α, IFN-γ) and cellular activation levels (anti-CD8+/CD38+ and CD3+/HLA-DR+)
were assessed. The onset of IRIS was assessed, being defined as a documented worsening of signs or symptoms of
TB during appropriate anti-tuberculosis treatment and after the initiation of highly active antiretroviral therapy
(HAART), not explained by any other disease or by an adverse effect of drug therapy. IRIS was observed for six
patients during the follow-up, within a mean time of 30 and 60 day. The individuals were then stratified into two groups
(No IRIS [= 48] and IRIS [n=6]) and the T cell activation subsets and the plasma concentration of pro and antiinflammatory cytokines were compared at baseline and 4 weeks after HAART introduction. Statistical analyses were
carried out using SPSS version 17.0. Increase of CD4 T cell counts and suppression of HIV viral load were observed
for the two groups. No statistically significant differences were observed for the T cell activation markers or the
cytokine panel used in the present study when baseline or 4 weeks after HAART were compared.
Conclusion: Our results indicate that HIV/TB patients under tuberculosis treatment and HAART are able to recover
the CD4+ T cell counts and control of viral replication. No specific prognostic biomarkers for IRIS were observed
based on the pro and anti-inflammatory cytokines and T cell activation subsets currently analyzed.
Financial support: DECIT CNPq
TCR VΒ REPERTOIRE DIVERSITY IN HEALTHY INDIVIDUALS OF DIFFERENT AGE GROUPS
JERUSA MARILDA ARANTES1; NATHÁLIA MOREIRA SANTOS2; MARCÍLIA SIERRO GRASSI3; ANA CRISTINA
AOUN TANNURI4; LUÍSA GUILHERME5; MAGDA CARNEIRO-SAMPAIO6.
1,3,4,6.DEPARTMENT OF PEDIATRICS, INSTITUTO DA CRIANÇA-HOSPITAL DAS CLÍNICAS DA FACULDADE
DE MEDICINA, SÃO PAULO - SP - BRASIL; 2,5.INSTITUTO DO CORAÇÃO, FACULDADE DE MEDICINA DA
UNIVERSIDADE DE SÃO PAULO, SÃO PAULO - SP - BRASIL.
Introduction. T cell receptors (TCRs) are expressed on the T cell surface serving for primary antigen recognition
function in adaptive immune responses. The repertoire of T lymphocytes in healthy individuals of different age groups
has not so far been deeply explored, and these information are essential not only for the comprehension of ontogeny
of immune response, but also as controls for evaluation of patients with immune disorders, such as primary and
secondary immunodeficiencies and autoimmune diseases. We investigated TCR Vβ repertoire in peripheral blood of
42 healthy infants, children, adolescents and adults through Vβ length spectratyping technique.
Methods and Results. The repertoire diversity was quantified utilizing the complexity score (CS), that represents the
sum of the number of peaks for each one of the 24 Vβ families. The parameters used for the analysis were distribution
profile of CDR3 (third complementarity determining region) peaks and the frequency of expression of each family. The
score of complexity were similar among the individuals independently of age. Most of the individuals expressed the 24
Vβ families, and presented TCR with non-Gaussian distribution polyclonal profiles.
Conclusions. These findings showed that TCR repertoire is fully developed/established since the first days of life.
Healthy infants and children presented repertoire diversity similar to healthy adults.
This research was supported by FAPESP (grants - grants 2008/58238-4 and 2011/21641-9), and by Fleury Medicina
Diagnóstica.
THE IFNΓ RECEPTOR SIGNALING PATHWAY PLAYS AN IMPORTANT ROLE IN THE IMMUNOMODULATION
EXERTED BY MESENCHYMAL STEM CELLS IN ACUTE KIDNEY INJURY
1
2
3
PRISCILLA BARBOSA COSTA ; MARINA BURGOS DA SILVA ; DANILO CANDIDO DE ALMEIDA ; PATRICIA
SEMEDO4; MARLENE ANTONIA DOS REIS5; ÁLVARO PACHECO SILVA6; NIELS OLSEN SARAIVA CAMARA7.
1,3.FEDERAL UNIVERSITY OF SÃO PAULO (UNIFESP), SÃO PAULO - SP - BRASIL; 2,4,7.UNIVERSITY OF SAO
PAULO (USP), SÃO PAULO - SP - BRASIL; 5.FEDERAL UNIVERSITY OF TRIÂNGULO MINEIRO (UFTM),
UBERABA - MG - BRASIL; 6.IIIEP, ALBERT EINSTEIN HOSPITAL, SÃO PAULO - SP - BRASIL.
Introduction: Acute kidney injury (AKI) is considered a progressive multifactorial injury. The use of mesenchymal
stem cells (MSC) is a promising therapy with great repair potential. For clinical success, it is important to consider the
MSC interaction with the microenvironment of inflammatory lesions (TNFα and IFNγ), which appears to potentiate the
therapeutic role of MSC. The present study evaluated the role of IFNγ in the activation of MSC in a model of acute
kidney injury.
Methods and Results: MSC from IFNγ receptor knockout animals (IFNR KO) and from wild type animals (WT
C57/Bl6) were isolated from adipose tissue of male mice. After phenotypic and functional characterization, both cells
were administered in mice with AKI. AKI was induced by clamping renal pedicles of male C57Bl/6 mice, for 45 min.
Four hours after reperfusion, IFNR KO or WT MSC were intraperitoneally administered, and 24 hours after surgery,
the animals were sacrificed. For all groups, n=8. Treatment with WT MSC showed significant improvement of renal
function parameters. Both treatments provided tissue regeneration (p<0,0001). The WT MSC group demonstrated
increased tissue proliferation (p<0,0001), and had lower rates of acute tubular necrosis, tissue apoptosis (p<0,05) and
oxidative stress (p<0,05) when compared with the IFNR KO MSC-treated or untreated groups. Analysis of the
inflammatory infiltrate showed a greater presence of leukocytes in the IFNR KO MSC-treated group when compared
to the WT MSC-treated or untreated groups (p<0,0001). Kidney genomic assays (RT-PCR) demonstrated that WT
MSC are more efficient in modulating the inflammatory response than IFNR KO MSC. In the WT MSC-treated group,
there was a decrease in the inflammatory response resulted from reperfusion, which was mainly evidenced by
changes in the Th1/Th2 balance, with a trend towards a Th2 type response. In addition, the gene level of expression
of renoprotective factors showed that WT MSC may promote more effective repair of kidneys after injury (p<0,05). The
expression ratio of anti-/pro-apoptotic molecules was also higher in the group treated with WT MSC (p<0,05).
Conclusion: The IFNγ receptor is important for MSC activation and has great importance in the immunomodulatory,
proliferative, cytoprotective and anti-apoptotic properties of these cells.
Financial support: grant 2011/14613-9, São Paulo Research Foundation (FAPESP), CNPq.
THE ROLE OF OBESITY AND CYTOKINES IL12/ IFN-G IN THE MODULATION OF ULCERATIVE COLITIS
INFLAMMATORY RESPONSE AND TREATMENT WITH SHORT CHAIN FATTY ACIDS
1
2
3
DENIS GONÇALVES SILVA ; ANGELA CASTOLDI ; FREDERICK WASINSKI ; VINICIUS DE ANDRADE
OLIVEIRA4; PEDRO M.M.M OLIVEIRA5; NIELS OLSEN SARAIVA CAMERA6.
1,3.UNIFESP, SAO PAULO - SP - BRASIL; 2,4,5,6.USP, SAO PAULO - SP - BRASIL.
(1) Departamento de Biofísica, Universidade Federal de São Paulo; (2) Departamento de Imunologia, Instituto de
Ciências Biomédicas, Universidade de São Paulo; (3) Departamento de Medicina, Universidade Federal de São
Paulo.
Introduction: The precise etiology of ulcerative colitis (UC) remains unclear, although it is believed that intestinal
inflammation results from dysregulated mucosal immune responses and a hypertrophy of mesenteric adipose, besides
a deficiency of short chain fatty acids (SCFA) may play an important role. Thus, this work aimed to study the role of
obesity and cytokines IL12/ IFNg in ulcerative colitis severity and possible mechanisms of the disease, as well as a
possible treatment with SCFA. Methods: Male mice, IL-12KO, IFN-gKO and WT, all C57BL/6J were used and obesity
induced with a high-fat diet (HF) (45% kcal from fat) for 8 weeks. After this time, UC were induced by 25g/L dextran
sulfate sodium (DSS) in water and the survival analyzed or sacrificed on the ninth day. On the final day was collected
serum (for circulating cytokine by Bioplex), colon tissue (histological analysis to gauge disease severity), and
mesenteric lymph nodes (FACS analysis of mucosal lymphoid tissue responses). For the study of SCFA, WT were
divided into groups with acetate or propionate or butyrate or without SCFA (CTL); all groups had 5 days of
pretreatment with its respective SCFA, diluted in water at a concentration of 150 mM or CTL, and also the same
concentration in the 40 days of induction of DSS colitis. Results: The groups WT HF and IFN-γ HF displayed higher
weight compared with their controls (p<0.001) and IL12KOs (p <0.001), on the other hand the knockout for IL-12 does
not gain weight compared to controls or the other two groups. The UC results showed that the group WT HF has lower
survival than the control animals (p<0.001) and knockouts for IL-12 (p<0.001) and IFN-γ (p<0.001). Also showed in
the same group a decrease in mesenteric lymph nodes in macrophages CD11b+F4/80+ (p<0.05) and CD4+ T cells
(p<0.05). And finally, the results revealed less weight loss (p<0.001) and a significant reducing of death in all groups
of SFCA, highlighting the butyrate group, which all survived the 40 days (CTL had a survival mean of 14
days). Conclusion: In WT obese mice, induced by diet, the low state of chronic systemic inflammation increases the
severity of colitis, and IL-12 and IFN-gKOs showed a lower severity of UC, demonstrating the importance of these proinflammatory cytokines in the disease. Moreover, the anti-inflammatory effects attributed to SFCA are actually doing
effect, protecting mice against the death and proved to be a promising treatment of UC.
Financial support: FAPESP
THE USE OF FUNCTIONAL STATUS CORRECTOR IN TREATMENT OF SALPINGO-OOPHORITIS
1
2
3
TATYANA YU. DANIELYAN ; NARINE A. DANIELYAN ; ANATOLIY P. GODOVALOV .
1,2.LTD. MEDICAL STUDIO, PERM - RÚSSIA; 3.ACAD. E.A. WAGNER PERM STATE MEDICAL ACADEMY, PERM
- RÚSSIA.
Introduction. Salpingo-oophoritis causes the changes in the menstrual, reproductive functions and secondary
functional changes in the nervous, endocrine, immune and other body systems. Nowadays new and more rational
modern methods of treatment are being developed. Russian scientist S.V. Koltsov and scientists of "Center Region"
have shown the therapeutic properties of the device for correction of functional status. This is a bioenergoinformational device which has no analogues in the world. Manufacturing technology of "Functional status
corrector" device (FSC) repeats the principles of electromagnetic fields organization at the cellular level in vivo. The
main directions of FSC are the structuring of liquids (RF Patent 32214843), correction of functional status of the
biological object (RF Patent 2214843), correction of external electromagnetic field affecting living organism (RF Patent
2262361). To use FSC the following documents are provided: registration certificate of Russian Ministry of Health
2008/02908 which indicates that FSC is a medical device; correspondence certificate № POCC RU.ИМ22.В01 703.
The effect of FSC on immune status (L.I. Radkova, 2012).
The purpose of the study is to analyze the clinical use of "Functional status corrector" (FSC) in treatment of of
salpingo-oophoritis as an effective addition to drug therapy.
Methods. The study group included 40 patients who were treated with medication therapy in combination with FSC antiparasitic (№ 1), detoxification (№ 2), corrector of female endocrine system (№ 3), rehabilitational (№ 5) in the form
of ingestion of structured water at regular use. The comparison group consisted of 20 people who received only
medical therapy.
Results. Before the treatment the reaction and pain syndrome was observed in 82% of the study group and in 85% of
the comparison group. Defect of cellular (80 and 82% respectively) and humoral component of immune system,
nonovulation (75 and 77%, respectively) was observed. During the treatment we observed elimination of inflammatory
reaction and pain syndrome (89 and 35% of patients, respectively), normalization of cell (82 and 34%, respectively)
and humoral component of immune system (88 and 32% of patients, respectively), the appearance of ovulatory cycles
(81 and 16 % of patients, respectively).
Conclusion. The use of FSC in combination treatment of salpingo-oophoritis is highly effective addition and can be
recommended for use in gynecological practice.
TLR8 ACTIVATION OVERCOMES THE IMPAIRED RESPONSIVENESS TO INTRACELLULAR TOLL-LIKE
PATHWAY IN LICHEN PLANUS
ROSANA DOMINGUES; GABRIEL COSTA DE CARVALHO; ELIANE AKEMI FUTATA; MARINA PASSOS
TORREALBA; LUANDA MARA DA SILVA DE OLIVEIRA; VALERIA AOKI; ALBERTO JOSÉ DA SILVA DUARTE;
MARIA NOTOMI SATO.
FACULDADE DE MEDICINA DA USP, SÃO PAULO - SP - BRASIL.
Introduction: Lichen planus (LP) is considered a chronic inflammatory mucocutaneous disease affecting 1-2% of the
population. LP is of unknown etiology probably of multifactorial origin, which raises the importance to study the innate
immune response to understand their chronic inflammatory status. Extracellular Toll-like receptor (TLR) senses
bacterial, fungal Pathogen Associated Molecular Patterns (PAMPs) while those intracellular TLRs senses for viral or
intracellular bacteria PAMPs. The aim of this study is to evaluate the profile of TLRs 2-9 activation on the proinflammatory cytokines responsiveness by peripheral blood mononuclear cells (PBMCs) of LP individuals.
Methods and Results: The study enrolled LP individuals (n=13, 12 female, 1 male, age range= 29-62) from
outpatient clinic of Dermatology Department, Hospital das Clínicas de São Paulo, and health individuals (n=10, 9
female, 1 male, age range= 24-57). PBMCs were cultivated in presence of agonists for TLRs, as PamCSK4 (TLR-2),
Poly (I:C) (TLR-3), lipopolysaccharide (LPS, TLR-4), flagelin (TLR-5), Imiquimod (TLR-7), CL097 (TLR-7/TLR8) or
CpG (TLR-9) for 48h. Supernatants were assessed for TNF, IL1-β, and IL-10 measurements using a Flex Set by
means of flow cytometry. The results showed enhanced TNF secretion upon extracellular TLR2, TLR4 and TLR5
activation in LP individuals compared to health control. In contrast, the IL-10 was reduced followed TLR-4 stimulation
in LP group. Upon intracellular TLRs activation, LP patients have remarked decrease in TNF and IL-1β secretion
levels via TLR7 activation. Notably, the compound CL097, ligand for TLR7/TLR8, overcomes the TNF secretion, but
not for IL-1β in LP individuals. In addition, LP individuals showed an increased IL-1β response for TLR9 stimulation.
Conclusion: The findings showed an altered profile of TLRs activation in LP individuals, with enhanced TNF
response for extracellular TLRs stimulation and absent response for intracellular TLR3 and TLR7 activation.
Interestingly, CL097 through TLR8, suggest an alternative pathway to revert impaired TNF secretion in LP.
Financial support: CNPq/FAPESP/LIM-56/HCFMUSP.
TREATMENT FOR PATIENTS CO-INFECTED WITH HIV AND VISCERAL LEISHMANIASIS
LÚSIO ARAÚJO LOPES JÚNIOR1; MARIANA CARLOS DE GÓIS2; GABRIELA LIMA NÓBREGA3; ATHENA
4
THAMIRIS DANTAS DE ALENCAR MARTINS QUEIROZ ; KARYNNE MARIA OLIVEIRA DA TRINDADE
5
6
MEDEIROS ; FERNANDA CARLOS DE GÓIS OLIVEIRA .
1,2,3,4,5.UNIVERSIDADE POTIGUAR, NATAL - RN - BRASIL; 6.CENTRO DE MEDICINA TROPICAL DE
RONDÔNIA, PORTO VELHO - RO - BRASIL.
Introduction: Visceral leishmaniasis (VL) widely known as kala-azar, is a systemic parasitic infection, primarily
affecting the reticuloendothelial system and is caused by the parasite Leishmania, which is transmitted by
hematophagous sand fly vector. The HIV epidemic has significantly influenced the epidemiology, clinical
manifestations and progression of VL. In return the VL also accelerates HIV progression by increasing viral replication,
leading to increased immunosuppression. Given this scenario, it is essential to the study of VL, especially in
immunocompromised patients, as this influences the response pattern of T lymphocytes, depressing macrophage
activation, further complicating the clinical condition of the patient with HIV.
Methods and Results: We performed a literature review on the topic co-infection visceral leishmaniasis and HIV. The
search for information occurred through access to review articles on electronic journals available online, such as
SciELO and Bireme. Treatment of visceral leishmaniasis in patients co-infected with HIV remains a challenge due to
poor response and toxicity of some drugs. Although antiretroviral therapy has resulted in dramatic reductions in the
incidence of co-infection with HIV in Southern Europe, it was found that has a partial protective effect against
outbreaks of VL. Relapse rate of 30% to 60% at one year were reported. Currently, secondary prophylaxis is given in
Europe after achieving parasitological cure. The administration of amphotericin B, pentavalent antimony and
pentamidine every 3 to 4 weeks has been used. However, experience with miltefosine for secondary prophylaxis is
limited.
Conclusion: The co-infection of HIV and visceral leishmaniasis is increasing worldwide, including in some regions of
Brazil like Rio Grande do Norte. Therefore, research efforts are needed for the institution of appropriate treatment and
drug administration more effective and which is adapted to changes in the host's immune system.
Financial support: This work did not receive financial support
VITAMIN D RECEPTOR POLYMORPHISMS AND VITAMIN D LEVELS: SUSCEPTIBILITY TO OSTEOPOROSIS IN
NORTHEAST BRAZILIAN PATIENTS?
1
2
JAQUELINE DE AZEVEDO SILVA ; ALEXANDRE DOMINGUES BARBOSA ; AMANDA LUIZE MELO TARAVARES
RAMOS3; CATARINA ADDOBBATI CAVALCANTI4; ANGELA LUIZA BRANCO PINTO DUARTE5; SERGIO
6
7
CROVELLA ; PAULA SANDRIN-GARCIA .
1,2,3.LABORATORY OF IMMUNEPATHOLOGY KEIZO ASAMI - LIKA, RECIFE - PE - BRASIL; 4,6,7.DEPARTMENT
OF GENETICS - UFPE, RECIFE - PE - BRASIL; 5.REUMATHOLOGY SERVICE FROM HOSPITAL DAS CLÍNICAS UFPE, RECIFE - PE - BRASIL.
Introduction: Osteometabolic Osteoporosis (OO) is a disorder characterized by diminished bone mineral density
(BMD) leading to an increased bone fragility and is mainly prevalent in postmenopausal women. Vitamin D is able to
increase calcium absorption by intestinal tract and acts on the deposition and bone reabsorption. The biological effect
of vitamin D is triggered from the connection between this hormone and specific cellular receptors - the vitamin D
receptor (VDR). Allelic variants in the VDR gene have been reported to account for changes in calcium homeostasis,
with subsequent effects on bone strength and size as well as in BMD. Therefore, the aim of this study was to
investigate whether polymorphisms in VDR gene are associated to BMD and the possible role of vitamin D levels in
diseases susceptibility. Methods and Results: In this study 156 OO female patients were enrolled, ranging from 5058 years old and 102 female healthy controls, from 41-67 years old, with the BDM ranging from -1 to -2.5 as an
exclusion criterion. Vitamin D levels were performed by ELISA for all patients. Genotyping was performed in 142
patients and 102 controls using the Taqman probes for the Tag-SNPs rs11168268, rs3890733 and rs1540339, which
encloses most of VDR gene by linkage disequilibrium (LD), and using Real time PCR 7500 (Applied Biosystems).
Genotype analysis followed manufacturer’s instructions and statistical analyses were performed using R software,
SNPassoc package. The results indicated no association to the tested SNPs, vitamin D levels and BMD in OO
Northeast Brazilian patients. Conclusion: VDR polymorphisms are not associated to vitamin D levels and BMD in
women with OO. However, more replica studies with a larger sample size are needed to confirm our findings. To our
knowledge this is the first association study enclosing VDR SNPs and Vitamin D levels in OO in Brazilian
patients. Financial support: FACEPE, CAPES and CNPq.