In silico prediction of peptides binding to multiple HLA-DR molecules accurately identifies immunodominant epitopes from gp43 of Paracoccidioides brasiliensis frequently recognized in primary peripheral blood mononuclear cell responses from sensitized individuals

One of the major drawbacks limiting the use of synthetic peptide vaccines in genetically distinct populations is the fact that different epitopes are recognized by T cells from individuals displaying distinct major histocompatibility complex molecules. Immunization of mice with peptide (181-195) from the immunodominant 43 kDa glycoprotein of Paracoccidioides brasiliensis (gp43), the causative agent of Paracoccidioidomycosis (PCM), conferred protection against infectious challenge by the fungus. To identify immunodominant and potentially protective human T-cell epitopes in gp43, we used the TEPITOPE algorithm to select peptide sequences that would most likely bind multiple HLA-DR molecules and tested their recognition by T cells from sensitized individuals. The 5 most promiscuous peptides were selected from the gp43 sequence and the actual promiscuity of HLA binding was assessed by direct binding assays to 9 prevalent HLA-DR molecules. Synthetic peptides were tested in proliferation assays with peripheral blood mononuclear cells (PBMC) from PCM patients after chemotherapy and healthy controls. PBMC from 14 of 19 patients recognized at least one of the promiscuous peptides, whereas none of the healthy controls recognized the gp43 promiscuous peptides. Peptide gp43(180-194) was recognized by 53% of patients, whereas the other promiscuous gp43 peptides were recognized by 32% to 47% of patients. The frequency of peptide binding and peptide recognition correlated with the promiscuity of HLA-DR binding, as determined by TEPITOPE analysis. In silico prediction of promiscuous epitopes led to the identification of naturally immunodominant epitopes recognized by PBMC from a significant proportion of a genetically heterogeneous patient population exposed to P. brasiliensis. The combination of several such epitopes may increase the frequency of positive responses and allow the immunization of genetically distinct populations.Univ Sao Paulo, Sch Med, Immunol Lab,Dept Med, Inst Heart,Incor,Div Clin Immunol & Allergy, BR-05403000 Sao Paulo, BrazilUniv Sao Paulo, Sch Med, Inst Invest & Immunol, Millenium Inst, BR-05403000 Sao Paulo, BrazilUniv Sao Paulo, Sch Med, Dept Infect & Parasit Dis, BR-05403000 Sao Paulo, BrazilLa Jolla Inst Allergy & Immunol, San Diego, CA USAHoffmann La Roche Inc, Dept Genomic & Informat Sci, Nutley, NJ 07110 USAUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Discipline Cell Biol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Microbiol Immunol & Parasitol, Discipline Cell Biol, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Biophys, Sao Paulo, BrazilWeb of Scienc