CHARACTERIZATION/MODULATION OF CYTOKINE RESPONSES IN FIV

本提案的目的是全面描述 淋巴组织中细胞因子mRNA和蛋白的表达 FIV感染的猫和试图调节细胞因子反应 用抗细胞因子抗体和重组李斯特菌 单核细胞增多症 假设：细胞因子产生不当 导致对FIV的无效免疫应答， FIV的起源，以及FIV感染猫对 机会性感染 细胞因子的调节可用于 研究治疗干预和疫苗的功效 策略，以及FIV的发病机制。 具体目标1： 确定FIV感染对细胞因子mRNA的影响， 猫组织中的蛋白质生成，多淋巴 将通过QC-RT-PCR评价隔室，以定量组织 细胞因子mRNA的产生，通过原位杂交定位 组织结构中的mRNA，通过免疫细胞化学， ELIspot和流式细胞术测定细胞因子蛋白 在特定的细胞表型中产生。 具体目标2： 确定IL 2、IL 4、IL 10、IL 12、TNF-α和 IFN-γ在FIV的免疫发病机制中，抗IL-2，抗IL-4， 抗-IL 10、抗-IL 12 p40、抗-TNF-α或抗-IFN-γ 将给予单克隆和多克隆抗体， 急性FIV感染的猫。 细胞因子抑制对 病毒复制、免疫应答和疾病进展将是 测定 具体目标3：确定是否存在强细胞介导的 对FIV免疫力有保护作用，SPF猫只将接受肝脏疫苗注射 重组生物疫苗载体; FIVenv-和/或FIVgag- 表达单核细胞增生李斯特菌。 猫将受到挑战 与同源和异源FIV和免疫应答 以及与保护性或非保护性 将评估保护性免疫力。 意义：慢病毒- 相关免疫缺陷是一个日益扩大的全球性问题 影响人类和猫，然而免疫发病机制是 不完全理解，目前的治疗方法有限。 了解细胞因子在发病机制中的作用， FIV感染的治疗和疫苗接种将直接 对艾滋病毒感染的影响。

The objective of this proposal is to comprehensively characterize cytokine mRNA and protein expression in the lymphoid tissues of FIV-infected cats and attempt to modulate the cytokine response with anti-cytokine antibodies and recombinant Listeria monocytogenes. Hypothesis: Inappropriate cytokine production contributes to the ineffective immune response to FIV, the patho- genesis of FIV, and the susceptibility of FIV infected cats to opportunistic infections. Modulation of cytokines may be useful to study the efficacy of therapeutic interventions and vaccine strategies, as well as the pathogenesis of FIV. Specific Aim 1: to determine the effect of FIV infection on cytokine mRNA and protein production in tissues of cats, multiple lymphoid compartments will be evaluated by QC-RT-PCR to quantify tissue production of cytokine mRNA, by in situ hybridization to localize mRNA within the tissue structure, and by immunocytochemistry, ELIspot, and flow cytometry to determine cytokine protein production within specific cell phenotypes. Specific Aim 2: to determine the significance of IL2, IL4, IL10, IL12, TNF-alpha and IFN-gamma in the immunopathogenesis of FIV, anti-IL2, anti-IL4, anti-IL10, anti-IL12p40, anti-TNF-alpha or anti-IFN-gamma monoclonal and polyclonal antibodies will be administered to acutely FIV infected cats. The effects of cytokine inhibition on viral replication, immune response, and disease progression will be determined. Specific Aim 3: To determine if a strong cell-mediated immunity to FIV is protective, SPF cats will be vaccinated with live recombinant biological vaccine vector; FIVenv- and/or FIVgag- expressing Listeria monocytogenes. Cats will then be challenged with homologous and heterologous FIV and the immune response and cytokine profile associated with either protective or non- protective immunity will be evaluated. Significance: Lentivirus- associated immunodeficiency is an expanding global problem affecting humans and cats, however the immunopathogenesis is incompletely understood and current therapies are limited. Understanding the role of the cytokines in the pathogenesis, therapy, and vaccination in FIV infection will have direct implications for HIV infection.