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

研究口服疫苗特异性组织驻留记忆 CD4 T 细胞的组织定位和保护功能

• 批准号: 10646930
• 负责人: Timothy Wesley Hand
• 金额: $ 6.39万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-02-01 至 2025-01-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10646930
• 关键词: Address; Adjuvant; Adoptive Transfer; Antigens; Attenuated; B-Lymphocytes; Bacteria; Binding; Biology; C57BL/6 Mouse; CD4 Positive T Lymphocytes; CD8B1 gene; CRISPR/Cas technology; Cell Communication; Cells; Child; Communicable Diseases; Communities; Congenic Mice; Cytoprotection; Development; Diarrhea; Dissection; Enteral; Enteric Nervous System; Escherichia coli; Flow Cytometry; Genes; Goals; Grant; Image; Immune; Immune response; Immunity; Immunize; Immunoglobulin A; Infection; Infection prevention; Intestines; Invaded; Listeria monocytogenes; Location; Longevity; Lymphoid Tissue; MHC Class II Genes; Measurement; Mediating; Memory; Memory B-Lymphocyte; Methods; Microscopy; Modeling; Mucous Membrane; Mus; Nerve; Oral; Peptides; Phenotype; Poliomyelitis; Population; Positioning Attribute; Preventive therapy; Production; Proliferating; Property; Proteins; Research; Research Personnel; Resource-limited setting; Resources; Rotavirus; SARS-CoV-2 variant; Site; Small Intestines; Small intestine mucous membrane; Spleen; Surface; System; T cell response; T memory cell; T-Cell Activation; T-Cell Receptor; T-Lymphocyte; Testing; Tissues; Toxin; Transgenic Mice; Transgenic Organisms; Vaccination; Vaccinee; Vaccines; Work; breakthrough infection; cell motility; congenic; cytotoxic; diarrheal disease; enteric infection; enteric pathogen; enterotoxigenic Escherichia coli; gut colonization; interest; memory CD4 T lymphocyte; mortality; mouse model; mucosal site; mutant; novel; oral vaccine; pathogen; pathogenic bacteria; pathogenic virus; prevent; protective effect; response; restoration; tissue resident memory T cell; tool; transcriptomics

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.

摘要 接种疫苗是我们预防传染病的最重要的方法。保护性免疫力. 粘膜部位具有通过阻止宿主入侵来防止再感染的潜力。小肠 粘膜是最常见的感染部位之一，但疫苗诱导的免疫机制， 在该位点的介导的保护仍然相对未知。有效疫苗的主要目标是 产生提供持久保护的长寿B细胞和T细胞。CD 4 + T细胞如何有助于 疫苗介导的保护作用尚不清楚。我们已经开发了一种口服疫苗接种模型， E.大肠杆菌热不稳定毒素（LT）诱导大量疫苗特异性CD 4 + T细胞， 是防止再次感染的必要条件。使用MHC II类四聚体，我们证明了口服 疫苗接种诱导LT特异性肠道记忆CD 4 + T细胞的长寿群体， 组织驻留记忆T细胞（Tissue-resident memory T cells，Trms）。LT特异性T细胞也表达基因 与肠神经功能相关，接种疫苗后，在肠神经功能附近观察到CD 4 + T细胞。 神经我们的假设是口服疫苗特异性CD 4 + Trms通过与神经相互作用来保护肠道 在重新遇到肠道病原体的抗原时激活运动性并以物理方式排出肠道病原体。为了验证这一 我们需要开发一种新的T细胞受体（TCR）转基因小鼠，这种小鼠对显性MHC具有特异性 通过LT特异性TCR转基因记忆T细胞的过继转移，我们可以 测试这些细胞足以介导对肠道感染的保护的假设。此外，LT- 特异性TCR转基因T细胞可以很容易地在组织中使用同源标记物鉴定，我们建议 用它们来确定口服疫苗激活的CD 4 + T细胞在小肠中与哪些神经相互作用。在 这项拨款的结论是，我们将开发出第一种TCR转基因小鼠， 提高我们的能力，使有关生物学和CD 4 + Trms的保护作用的重要发现， 小肠，而且毫无疑问也将用于对口服疫苗感兴趣的科学界。