Braga Catarina J M, Rittner Glauce M G, Muñoz Henao Julian E, Teixeira Aline F, Massis Liliana M, Sbrogio-Almeida Maria E, Taborda Carlos P, Travassos Luiz R, Ferreira Luís C S
Department of Microbiology-ICB, University of São Paulo, Av. Prof. Lineu Prestes 1374, São Paulo, SP 05008-000, Brazil.
Infect Immun. 2009 Apr;77(4):1700-7. doi: 10.1128/IAI.01470-08. Epub 2009 Feb 9.
Paracoccidioidomycosis (PCM) is a systemic granulomatous disease caused by the dimorphic fungus Paracoccidioides brasiliensis. Anti-PCM vaccine formulations based on the secreted fungal cell wall protein (gp43) or the derived P10 sequence containing a CD4(+) T-cell-specific epitope have shown promising results. In the present study, we evaluated new anti-PCM vaccine formulations based on the intranasal administration of P. brasiliensis gp43 or the P10 peptide in combination with the Salmonella enterica FliC flagellin, an innate immunity agonist binding specifically to the Toll-like receptor 5, in a murine model. BALB/c mice immunized with gp43 developed high-specific-serum immunoglobulin G1 responses and enhanced interleukin-4 (IL-4) and IL-10 levels. On the other hand, mice immunized with recombinant purified flagellins genetically fused with P10 at the central hypervariable domain, either flanked or not by two lysine residues, or the synthetic P10 peptide admixed with purified FliC elicited a prevailing Th1-type immune response based on lung cell-secreted type 1 cytokines. Mice immunized with gp43 and FliC and intratracheally challenged with P. brasiliensis yeast cells had increased fungal proliferation and lung tissue damage. In contrast, mice immunized with the chimeric flagellins and particularly those immunized with P10 admixed with FliC reduced P. brasiliensis growth and lung damage. Altogether, these results indicate that S. enterica FliC flagellin modulates the immune response to P. brasiliensis P10 antigen and represents a promising alternative for the generation of anti-PCM vaccines.
副球孢子菌病(PCM)是一种由双相真菌巴西副球孢子菌引起的全身性肉芽肿疾病。基于分泌型真菌细胞壁蛋白(gp43)或含有CD4(+) T细胞特异性表位的衍生P10序列的抗PCM疫苗制剂已显示出有前景的结果。在本研究中,我们在小鼠模型中评估了基于鼻内给予巴西副球孢子菌gp43或P10肽并联合鼠伤寒沙门氏菌FliC鞭毛蛋白(一种与Toll样受体5特异性结合的天然免疫激动剂)的新型抗PCM疫苗制剂。用gp43免疫的BALB/c小鼠产生了高特异性血清免疫球蛋白G1反应,并提高了白细胞介素-4(IL-4)和IL-10水平。另一方面,用在中央高变域与P10基因融合的重组纯化鞭毛蛋白免疫的小鼠,无论其两侧是否有两个赖氨酸残基,或者用与纯化的FliC混合的合成P10肽免疫的小鼠,基于肺细胞分泌的1型细胞因子引发了主要的Th1型免疫反应。用gp43和FliC免疫并经气管内接种巴西副球孢子菌酵母细胞的小鼠,真菌增殖和肺组织损伤增加。相比之下,用嵌合鞭毛蛋白免疫的小鼠,特别是用与FliC混合的P10免疫的小鼠,巴西副球孢子菌的生长和肺损伤减少。总之,这些结果表明鼠伤寒沙门氏菌FliC鞭毛蛋白可调节对巴西副球孢子菌P10抗原的免疫反应,是生成抗PCM疫苗的一种有前景的替代方法。