Dendritic cells & immunity in mycoplasma pneumonia

描述（由申请人提供）：本项目的长远目标是发展预防和治疗呼吸系统疾病的方法。我们发现不同的T细胞群调节小鼠肺炎支原体的炎症反应，而决定T细胞反应类型的因素需要被理解。树突状细胞（dc）和其他抗原呈递细胞（如巨噬细胞）在支原体疾病中的作用尚不清楚，但由于T细胞反应在支原体疾病中起着至关重要的作用，因此树突状细胞和巨噬细胞可能影响疾病的发病机制并影响保护性免疫的产生。我们假设，控制DC可以预测地改变肺中针对支原体感染的免疫反应的类型和强度，这将影响疾病的严重程度和抵抗力。此外，肺巨噬细胞也可能影响支原体免疫，可能是通过支持th1型反应。需要比较肺dc和巨噬细胞活性的影响。在本提案中，我们将解决以下问题：1)来自幼稚或支原体感染小鼠的肺dc和巨噬细胞的变化是否影响激活和调节T细胞分化和激活的能力？2)支原体抗原脉冲dc或巨噬细胞能否对支原体产生保护性或免疫病理反应？3)调节细胞因子改变DC功能是否影响肺免疫反应和支原体呼吸系统疾病？4) dc -趋化因子ABCD-1和ABCD-2 （TARC）是否在支原体肺免疫中起作用？实验设计如下：1)研究支原体感染小鼠呼吸道dc细胞和巨噬细胞在体外和体内对无关抗原（OVA）激活T细胞应答的能力。2)用支原体抗原脉冲树突状细胞和巨噬细胞，并测定它们在气管内接种支原体和随后的支原体攻击后产生耐药性或免疫病理的能力。3)体外用调节性细胞因子、IFNgamma、IL-12、IL-4、tgf - β或IL-10，支原体抗原脉冲处理DC，通过气管内接种检测其对支原体产生耐药性或免疫病理的能力；4)检测支原体感染小鼠肺部T细胞中ABCD-1和ABCD-2受体的表达及其对这些趋化因子的反应能力。小鼠将被中和单克隆抗体处理，以确定ABCD-1和ABCD-2在支原体的保护性免疫和免疫病理反应中的作用。

DESCRIPTION (provided by applicant): The broad, long-term objectives of this project are to develop approaches to prevent and treat respiratory diseases. We found that different T cells populations modulate inflammatory responses of murine mycoplasma pneumonia, and the factors that determine the type of T cell responses need to be understood. The role of dendritic cells (DCs) and other antigen presenting cells, e.g. macrophages, in Mycoplasma disease is unknown, but because T cell responses in mycoplasma disease play such a critical role, DCs and macrophages likely impact on disease pathogenesis and influence the generation of protective immunity. We hypothesize that manipulating DC can predictably change the types and intensity of immune reactions in the lung against mycoplasma infection, and this will impact on the severity and resistance to disease. In addition, pulmonary macrophages may also impact on mycoplasma immunity, possibly through support of Th1-type responses. The impact of pulmonary DCs and macrophages activity needs to be compared. In this proposal, we will address the following questions: 1) Do changes in pulmonary DCs and macrophages from naive or mycoplasma-infected mice influence the ability to activate and modulate T cell differentiation and activation? 2) Can mycoplasma-antigen pulsed DCs or macrophages generate protective or immunopathologic responses against mycoplasma? 3) Can altering DC function with regulatory cytokines influence pulmonary immune responses and mycoplasma respiratory disease? 4) Do the DCbeta-chemokines, ABCD-1 and ABCD-2 (TARC), play a role in pulmonary immunity against mycoplasma? The experimental design is as follows: 1) DCs and macrophages from the respiratory tracts of mycoplasma-infected mice will be evaluated for their ability activate in vitro and in vivo T cell responses against an unrelated antigen (OVA). 2) DCs and macrophages will be pulsed with mycoplasma antigen and their ability to generate resistance or immunopathology after intratracheal inoculation and subsequent challenge with mycoplasma will be determined. 3) DC will be treated in vitro with regulatory cytokines, IFNgamma, IL-12, IL-4 TGF-beta or IL-10, and pulsed with mycoplasma antigen, and their ability to generate resistance or immunopathology against mycoplasma will be determined by their intratracheal inoculation; and 4) T cells from lungs of Mycoplasma infected mice will be examined for expression of receptors for ABCD-1 and ABCD-2 and their ability to respond to these chemokines. Mice will be treated with neutralizing monoclonal antibodies to determine the role of ABCD-1 and ABCD-2 in protective immunity and immunopathologic responses against mycoplasma.