Priming of Antifungal T-Cells at Mucosal and Systemic Sites

粘膜和全身部位抗真菌 T 细胞的启动

• 批准号: 7359255
• 负责人: Marcel Wuethrich
• 金额: $ 18.38万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-12-15 至 2009-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7359255
• 关键词: Activation Analysis; Adoptive Transfer; Antifungal Agents; Antigens; Attenuated; Attenuated Live Virus Vaccine; Autologous; Blastomyces; Blastomyces dermatitidis; CD4 Positive T Lymphocytes; Cells; Communicable Diseases; Complex; Data; Development; Disease; Effector Cell; Engineering; Event; Evolution; Experimental Models; Generations; Genital system; Hand; Immune; Immune response; Immunity; Immunization; Immunology; Infection; Infection Control; Infectious Agent; Influenza; Knowledge; Lead; Lung; Lung diseases; Mediating; Microbe; Molecular; Monitor; Mucosal Immunity; Mucous Membrane; Mus; National Institute of Allergy and Infectious Disease; Papillomavirus; Patients; Phase; Poliomyelitis; Property; Reagent; Research; Resistance; Respiratory Mucosa; Respiratory Tract Infections; Rotavirus; Route; Site; Skin; Staging; Surface; System; T-Cell Receptor; T-Lymphocyte; Testing; Tissues; Today; Transgenic Mice; Transgenic Organisms; United States National Institutes of Health; Vaccination; Vaccines; Work; base; design; fungus; improved; in vivo; insight; lymph nodes; microbial; mucosal vaccination; mucosal vaccine; pathogen; prevent; respiratory; response; subcutaneous; tool

DESCRIPTION (provided by applicant): Fungal pathogens and most infectious agents enter the body at mucosal surfaces. Mucosal immune priming induces both systemic and mucosal resistance, and is believed to be the ideal strategy to prevent or treat infections that arise from a mucosal surface. Systemic priming, in turn, usually induces poor mucosal immunity and is judged to be less desirable against mucosal infections. Contrary to this paradigm, we find that a live-attenuated, vaccine strain of Blastomyces dermatitidis can protect mice against a lethal pulmonary infection (via Th1 CD4+ T cells) when the attenuated strain is delivered only by the subcutaneous route, and not by the respiratory route. Here, we will investigate the fundamental mechanisms by which these 2 routes of antigen (Ag) delivery trigger CD4+ T cells during the priming and the recall phases. We will endeavor to uncover the cellular and molecular mechanisms that lead to effective or failed priming of T cells in the skin versus the lung. T cell receptor (TCR) transgenic (Tg) systems are arguably the best immunological tools to decipher the activation of clonal T cells in vivo, but there are no such reagents available for studying T-cell immunity to the dimorphic fungi. Therefore, we propose to create a Blastomyces-specific CD4+ TCR Tg mouse to monitor and analyze the activation of Ag-specific CD4+ T cells in these two immunologically disparate settings. We hypothesize that the subcutaneous delivery of Ag will prime TCR Tg, Blastomcyes-specific CD4+ T-cells in the skin-draining lymph nodes to become activated Th1 effector cells, and protect against lung challenge upon recall. Mucosal priming, on the other hand, will lead to the generation of functionally impaired Th1 effectors unable to provide resistance to challenge. Our specific aims are to: 1) Develop an adoptive transfer system to monitor Blastomyces-specific CD4+ T cells in vivo. Fungal-specific TCR Tg mice will be created from a T cell clone that protects mice against experimental infection. TCR Tg cells will be used to establish an autologous adoptive transfer system to track the phenotypic and functional properties of protective Ag-specific T cells during the priming phase in the skin, and on recall in the lung. 2) Identify whether and how mucosal priming fails to engender Ag-specific Th1 CD4+ cells. TCR Tg cells will let us track the critical stages of the CD4+ T cell response after Ag exposure at the respiratory mucosa: recruitment, activation, proliferation and differentiation in the lung and draining lymph nodes during the priming and recall phases. These events will be compared/contrasted to those after Ag exposure in the skin. Our studies will yield new insight into the evolution of T cell responses and immunity in mucosal tissues. This knowledge will contribute to the development of effective strategies to induce acquired, protective immunity for improved control of fungal and other microbial infections at these sites. PROJECT NARRATIVE: Fungal and other pathogens enter the body through the lung and cause disease at this site or after dissemination. The studies proposed here will create a uniquely powerful transgenic mouse to investigate the induction of protective immune responses in the lung. Our findings will provide the mechanistic basis to design strategies that prevent and/or treat infections with fungi and other microbial infections.

描述（由申请方提供）：真菌病原体和大多数传染性病原体从粘膜表面进入体内。粘膜免疫引发诱导全身性和粘膜抗性，并且被认为是预防或治疗由粘膜表面引起的感染的理想策略。而全身性预充通常诱导不良的粘膜免疫，并且被认为对粘膜感染不太理想。与这种模式相反，我们发现，当减毒菌株仅通过皮下途径而不是通过呼吸途径递送时，皮肤炎芽生菌减毒活疫苗株可以保护小鼠免受致死性肺部感染（通过Th 1 CD 4 + T细胞）。在这里，我们将调查的基本机制，这2个途径的抗原（Ag）交付触发CD 4 + T细胞在启动和召回阶段。我们将奋进揭示导致皮肤与肺中T细胞有效或失败的细胞和分子机制。T细胞受体（TCR）转基因（Tg）系统可以说是最好的免疫学工具，破译克隆T细胞在体内的激活，但有没有这样的试剂可用于研究T细胞免疫的二型真菌。因此，我们建议建立一个芽生菌特异性的CD 4 + TCR Tg小鼠，以监测和分析在这两个免疫学上不同的设置Ag特异性的CD 4 + T细胞的激活。我们假设，皮下递送Ag将引发皮肤引流淋巴结中的TCR Tg、母细胞瘤特异性CD 4 + T细胞成为活化的Th 1效应细胞，并在回忆时保护肺免受攻击。另一方面，粘液素引发将导致产生功能受损的Th 1效应物，其不能提供对攻击的抗性。我们的具体目标是：1）开发一种过继转移系统，以监测体内芽生菌特异性CD 4 + T细胞。真菌特异性TCR Tg小鼠将由保护小鼠免受实验感染的T细胞克隆产生。TCR Tg细胞将用于建立自体过继转移系统，以在皮肤中的致敏阶段和在肺中的回忆期间追踪保护性Ag特异性T细胞的表型和功能特性。2)确定粘膜引发是否以及如何未能产生Ag特异性Th 1 CD 4+细胞。TCR Tg细胞将使我们能够追踪呼吸道粘膜接触银后CD 4 + T细胞反应的关键阶段：在启动和回忆阶段，肺和引流淋巴结中的募集、激活、增殖和分化。将这些事件与皮肤中Ag暴露后的事件进行比较/对比。 我们的研究将产生新的见解的T细胞反应和免疫粘膜组织的演变。这些知识将有助于开发有效的策略，以诱导获得性保护性免疫力，从而改善对这些部位真菌和其他微生物感染的控制。 项目叙述：真菌和其他病原体通过肺部进入人体，并在该部位或传播后引起疾病。本文提出的研究将创造一种独特的强大的转基因小鼠，以研究肺中保护性免疫反应的诱导。我们的研究结果将为设计预防和/或治疗真菌感染和其他微生物感染的策略提供机制基础。