Investigating the tissue location and protective function of oral vaccine-specific tissue resident memory CD4 T cells

摘要 接种疫苗是我们预防传染病的最重要的预防方法。保护性免疫力 粘膜部位有可能通过阻止宿主的入侵来防止再次感染。小肠病 粘膜是最常见的感染部位之一，但疫苗诱导免疫的机制是- 这个部位的介导性保护仍然相对未知。一种有效疫苗的主要目标是 创造长寿的B细胞和T细胞，提供持久的保护。CD_4~+T细胞如何参与 疫苗介导的保护作用尚不清楚。我们已经开发出一种口服疫苗接种模式，该模式具有减毒效果 大肠杆菌不耐热毒素(LT)的一种版本，它能诱导大量疫苗特异性的CD4+T细胞 是防止再次感染所必需的。使用MHC II类四聚体，我们证明了口头 疫苗接种诱导长寿的LT特异性肠道记忆CD4+T细胞共享 组织驻留记忆T细胞(TRMS)的转录切割签名。LT特异性T细胞也表达基因 与肠神经功能有关，接种后可在肠道旁观察到CD4+T细胞 太紧张了。我们的假设是，口服疫苗特异性的CD4+TRMS通过与神经相互作用来保护肠道 当再次遇到肠道病原体的抗原时，激活运动并物理地将其排出。为了测试这一点 问题：我们需要开发一种新型的T细胞受体(TCR)转基因小鼠，专用于优势MHC 来自LT的II类限制性抗原。通过过继转移LT特异性的TCR转基因记忆T细胞，我们可以 验证这样的假设，即这些细胞足以介导对肠道感染的保护。此外，LT- 利用同源标记可以很容易地在组织中识别特定的TCR转基因T细胞，我们建议 使用它们来识别口服疫苗激活的CD4+T细胞在小肠中与哪些神经相互作用。在 这笔赠款结束后，我们将开发出第一只TCR转基因小鼠，它将极大地 提高我们在CD_4~+T细胞亚群的生物学和保护作用方面的重大发现的能力 小肠，也无疑会对科学界感兴趣的口服疫苗有所帮助。

Abstract Vaccination is our most important preventative therapy against infectious disease. Protective immunity at mucosal sites has the potential to prevent re-infection by stopping invasion of the host. The small intestinal mucosa is amongst the most common sites of infection, but the mechanisms of vaccine-induced immune- mediated protection at this site remain relatively unknown. The primary target of an effective vaccine is the creation of long-lived B cells and T cells that provide durable protection. How CD4+ T cells might contribute to vaccine-mediated protection is unclear. We have developed a model of oral vaccination with an attenuated version of the E. coli heat labile toxin (LT) that induces large populations of vaccine-specific CD4+ T cells that are necessary for protection against re-infection. Using MHC class II tetramers we demonstrated that oral vaccination induces a long-lived population of LT-specific intestinal memory CD4+ T cells share a transcriptomic signature with Tissue-resident memory T cells (Trms). LT-specific T cells also expressed genes associated with enteric nerve function and after vaccination CD4+ T cells were observed adjacent to enteric nerves. Our hypothesis is that oral vaccine-specific CD4+ Trms protect the intestine by interacting with nerves to activate motility and physically expel enteric pathogens upon re-encountering their antigen. To test this question we need to develop a novel T cell receptor (TCR) transgenic mouse, specific to a dominant MHC class II-restricted antigen from LT. Via adoptive transfer of LT-specific TCR transgenic memory T cells we can test the hypothesis that these cells are sufficient to mediate protection against enteric infection. Further, LT- specific TCR transgenic T cells can be easily identified in tissues using congenic markers and we propose to use them to identify which nerves oral vaccine-activated CD4+ T cells interact with in the small intestine. At the conclusion of this grant we will have developed a first in its kind TCR transgenic mouse that will drastically increase our ability to make important discoveries about the biology and protective effects of CD4+ Trms in the small intestine, but also will undoubtedly be of use to the scientific community interested in oral vaccines.