28ª REGEM
Reunião de Genética de Microrganismos
09 a 11 de setembro de 2012 - Rafain Palace Hotel & Convention Center - Foz do Iguaçu - PR
Protective effect of enolase DNA vaccine in candidiasis
Soares, DA1; Mingori, MR1,2 ; Silveira, CP1,2; Fuentefria, AM3; Vainstein, MH1,2.
CBiot - Centro de Biotecnologia, UFRGS, Porto Alegre, RS.
PPGBCM – Programa de Pós-graduação em Biologia Celular e Molecular, UFRGS.
³ Departamento de Análises, Faculdade de Farmácia, UFRGS.
1
2
[email protected]
Keywords: Candida, DNA vaccine, enolase, preventive vaccination and immune response
Candida albicans and non-albicans are opportunistic yeasts that cause both mucocutaneous and systemic candidiasis and
are important agents of infection mainly in immunocompromised patients. It is the fungal species most frequently isolated
in hospital-acquired infections and incidences of infection have risen with the increased prevalence of immunosuppressive
therapies and broad-spectrum antibiotic use. Disseminated candidiasis is associated with high mortality and drug resistance.
The treatment of this infection is ineffective in several cases; therefore the development of an anti-candidiasis vaccine
offers a preventive approach to reduce the incidence of this disease. Plasmid DNA vaccination may offer several important
advantages over traditional vaccines, e.g. the relative stability of DNA, the specificity of the antigen produced, and the
possibility of guiding the type of elicited specific immune response. In this study, our attention is focused on the highly
conserved glycolytic enzyme enolase. Enolase(2-phospho-D-glycerate hydrolyase, EC 4.2.1.11), catalyses a reversible
conversion of 2-phosphoglycerate to phosphoenolpyruvate. This protein is located on the cell wall of C. albicans and is
highly immunogenic. The objective of this study is to assess the effect of the preventive vaccination by DNA vaccine that
expresses the enolase protein of C. albicans. Enolase cDNA was cloned from a clinical isolate of C. albicans (ATCC 18804),
into eukaryotic expression pCDNA3.1 plasmid (Invitrogen). The transient expression (enolase::pCDNA) was performed
in lung epithelial cells A549. The enolase expression was confirmed by PCR using specific primers for enolase gene. The
prospects of this study are the phagocytosis assay with RAW- 264.7 macrophages and in vivo infection and immunization.
Mice will be immunized with the recombinant plasmid and posteriorly infected with clinical isolate of C. albicans (ATCC
18804). Cytokines (IL-10, TNF-α and IFN- γ) will be determined using the ELISA method.
Financial Support: CNPq and FINEP
MICRO040