CPAF induced CD4+ T cell mediated immunity against Chlamydia

描述(申请人提供)：目前还没有针对沙眼衣原体的许可疫苗，沙眼衣原体是全球细菌性传播疾病的主要原因。未经治疗的生殖器衣原体感染会导致严重的后遗症，如盆腔炎、宫外孕和不孕症。衣原体分泌的蛋白，命名为CPAF(衣原体蛋白酶/蛋白酶体样活性因子)，负责降解宿主MHC转录因子RFX5和USF1。因此，抑制CPAF活性将是阻止衣原体逃避免疫识别的一种可行的疫苗接种策略。我们现在已经提供了直接证据来证明在鼻腔(I.N.)中使用重组(R)CPAF和IL-12的这种方法的有效性。传送系统。鼻腔接种rCPAF和IL-12可诱导抗原特异性干扰素-3的产生，显著减少细菌在阴道内的脱落。与模拟免疫(PBS)小鼠相比，沙眼衣原体攻击显著加快了感染的消退。重要的是，接种rCPAFIL-12的小鼠对衣原体感染的病理后果表现出保护作用，包括输卵管系膜炎症以及输卵管积水和输卵管扩张。RCPAFIL-12介导的衣原体感染的溶解和抗炎性病变的保护高度依赖于内源性干扰素-3的产生和抗原特异性的CD4T细胞的作用。根据我们的大量证据，我们假设“CPAF疫苗通过在局部吞噬细胞的参与下，通过抗原特异性的干扰素-3分泌的CD4T细胞，提高上生殖道内的细菌清除能力，从而促进生殖器衣原体感染后的生育能力的保存和减少炎症病理”。我们将通过以下方式检验这一假设：(1)检验I.N的直接效应。(2)确定衣原体疫苗接种后衣原体生物减少与上生殖道内抗原特异性CD4T细胞浸润之间的关系；(3)研究干扰素-3产生CPAF特异性CD4T细胞增强细菌清除，防止与衣原体感染相关的泌尿生殖系统病理发展的机制(S)。公共卫生相关性：目前还没有针对沙眼衣原体的许可疫苗，沙眼衣原体是全球细菌性传播疾病的主要原因。使用衣原体感染的小鼠模型，鼻腔接种(重组衣原体蛋白酶/蛋白酶体样活性因子)CPAF和IL-12可诱导强大的抗原特异性干扰素-3的产生，显著减少细菌脱落，并诱导对衣原体感染的病理后果的保护，包括输卵管系膜炎症和输卵管积水和输卵管扩张。这项建议将进一步检查CPAF疫苗预防不孕不育的效果，并提供对这种保护性免疫性质的机械性洞察。

DESCRIPTION (provided by applicant): There currently is no licensed vaccine against Chlamydia trachomatis, the leading cause of bacterial sexually transmitted disease worldwide. Untreated genital chlamydial infection cause serious sequelae such as pelvic inflammatory disease, ectopic pregnancy, and infertility. A Chlamydia-secreted protein, designated as CPAF (chlamydial protease/proteasome-like activity factor), is responsible for degradation of the host MHC transcription factors RFX5 and USF1. Inhibiting CPAF activity will therefore be a feasible vaccination strategy for blocking chlamydial evasion of immune recognition. We have now provided direct evidence to demonstrate the effectiveness of such an approach using recombinant (r)CPAF and IL-12 in an intranasal (i.n.) delivery system. Intranasal vaccination with rCPAF and IL-12 induced robust antigen-specific IFN-3 production, significantly reduced bacterial shedding upon intravaginal (i.vag.) chlamydial challenge and significantly accelerated the resolution of infection compared to mock-immunized (PBS) mice. Importantly, rCPAF+IL-12 vaccinated mice exhibited protection against pathological consequences of chlamydial infection, including mesosalpingeal inflammation and development of hydrosalpinx and oviduct dilation. The rCPAF+IL-12-mediated resolution of chlamydial infection and protection against inflammatory pathology was highly dependent on endogenous IFN-3 production and the action of antigen-specific CD4+ T cells. Based on our notable body of evidence, we hypothesize that "CPAF vaccination promotes preservation of fertility and reduces inflammatory pathology after genital Chlamydia infection by enhancing bacterial clearance within the upper genital tract via antigen-specific IFN-3 secreting CD4+ T-cells with the participation of local phagocytic cells". We will test this hypothesis by; (1) Examining the direct effect of i.n. CPAF vaccination on the preservation of fertility after genital chlamydial challenge; (2) Determine the relationship between the reduction in chlamydial organisms and the infiltration of antigen-specific CD4+ T cells within the upper genital tract induced by CPAF vaccination; (3) Investigating the mechanism(s) by which IFN-3 producing CPAF specific CD4+ T cells enhance bacterial clearance and prevent the development of urogenital pathology associated with chlamydial infection. PUBLIC HEALTH RELEVANCE: There currently is no licensed vaccine against Chlamydia trachomatis, the leading cause of bacterial sexually transmitted disease worldwide. Using a murine model of genital chlamydial infection, intranasal vaccination with (recombinant chlamydial protease/proteasome-like activity factor) CPAF and IL-12 induced robust antigen-specific IFN-3 production, significantly reduced bacterial shedding and induced protection against pathological consequences of chlamydial infection, including mesosalpingeal inflammation and development of hydrosalpinx and oviduct dilation. This proposal will further examine the efficacy of CPAF vaccination against the prevention of infertility and provide mechanistic insight into the nature of this protective immunity.