2ª Jornada Fluminense sobre Cognição Imune e Neural
Os sinais de perigo: distinção entre o próprio e o não próprio
pelo sistema imune inato
ANM, Agosto de 2011
Marcello A. Barcinski
Uma história dos modelos imunológicos
and
Scand. J. Immunol. 54:2001
TLRs são PRRs
Superposições no reconhecimento
PAMPs e DAMPs
(PAMPS)
TLRs are PRRs:recognition system codified in the genome
(innate immune system)
Piccinini & Midwood. Mediators of Inflammation, 2010
Piccinini & Midwood. Mediators of Inflammation, 2010
Piccinini & Midwood. Mediators of Inflammation, 2010
Piccinini & Midwood. Mediators of Inflammation, 2010
DAMPs e doenças humanas
Piccinini & Midwood. Mediators of Inflammation, 2010
PAMPs e DAMPs
Resistance and Tolerance to Infectious Diseases
Schnaider and Ayres
2008
Variação da anemia (A) e da perda ponderal (B) em um modelo de malária murina
Science: 318, 2007
Seixas et al. PNAS 2009
Larsen et al. Science Transl. Med 2010
Heme Activates Innate Immunity
Figueiredo RT et al. Jour. Biol.Chem. 2007
Resistance and Tolerance to Infectious Diseases
Inflamação e anti-inflamação
Blander M.
Morte Celular sem Inflamação
Células e corpos apoptóticos são eficientemente fagocitados
Reconhecimento de PS
Immunosuppressive
and
anti-inflammatory signals
• TGF-β
β
• IL-10
Exposição de PS pode ser detectada pela ligação com Anexina-5
PS exposure detected by fluorescence
(Annexin V binding)
Promastigotes
Amastigotes
Leishmania life-cycle
PS exposure by amastigotes of Leishmania (L) amazonensis
Control
Dig. +Annex. + PI
PI
Annexin V-FITC
Annex. + PI
Marcação de amastigotas com anti-PS
Identidade no reconhecimento de células apoptóticas e patógenos
Stuart&Ezekowitz
Immunity, 2005:539
Mimetismo apoptótico : parasitas travestidos de próprio apoptótico
Gregory & Devitt, Immunology 2004
Apoptotic mimicry: masking PAMPs with ACAMPs (a
co
co--evolutionary acquisition for a parasitic lifestyle)
TGF-β1 production by
macrophages
Intracellular
digestion
PS exposing
amastigotes
TGF-β1 production by
macrophages
Intracellular
proliferation
By Fernando Real
PS exposing
apoptotic cell
Differential PS Exposure
(PS exposure is modulated by the host ?)
PI
PI + AX
∆MFI
MFI=26.94
=26.94
Propidium Iodide (PI)
Amastigotes
purified from
BALB/c mice
PI
PI + AX
∆MFI
MFI=7.35
=7.35
Amastigotes
purified from
C57Bl/6 mice
Annexin VV-FITC (AX)
Density of exposed PS moieties correlates with in vitro and in vivo
amastigote’s infectivity
BALB/c
C57Bl/6
Wanderley, JLM e cols. J. Immunol. 2006, 176:1834
T-cell-deficient mice expose less PS than WT mice
Protocol
Infect with Leishmania amazonensis;
Collect lymph-nodes;
Purify and characterize T-cells;
Transfer to infected nude mice;
Collect amastigotes and measure PS exposure.
Transfer of CD4+ but not of CD8+ T-cells induces PS exposure in amastigotes
infecting Balb/c nu/nu mice
Wanderley et al. submitted
Persistence of a mixed Th1 and Th2 response in the course
of infection with Leishmania amazonensis
Soong L. Amer. J. Trop. Med. Hyg. 2002
Varki A. PNAS, 2010
McFall-Ngai, M. Nature 2007
McFall-Ngai, M. Nature 2007
A Moment of Evolutionary Ecstasy Bought Us 400 Million Years
of Misery
Hedrick S. Immunity, 2004
Mechanism for uptake of apoptotic cells involves a combination of tethering
and signaling receptors
Henson et al., Curr. Biol., 11:R795-R805, 2001
Macropinocitose como Mecanismo de
Internalização de Formas Amastigotas
Wanderley, JLM e cols. J. Immunol. 2006, 176:1834
Synthesis of active TGFTGF-β1, induction of IL
IL--10 message and inhibition of NO
synthesis by macrophages correlate with the amount of surface PS displayed by
infective amastigotes
BALB/c
C57Bl/6
MO
Wanderley et al., J. Immunol., 176:1834-1839, 2006
PS exposure is modulated by the host ?
Macrophages of different genetic origins do not
induce differential PS exposure
Wanderley et al. submitted
Protocol
Infect with Leishmania amazonensis;
Collect lymph nodes;
Purify lymphocytes;
Generate immune supernatants by activating with SLA.
Immune T cells and T-cell supernatants induce PS exposure by amastigotes
Wanderley et al. submitted
SNs do not induce parasite death
Resistance and Tolerance to Infectious Diseases
Science: 318, 2007
Larsen R. et al. Sci. Transl. Med. 2010