CHARACTERIZATION/MODULATION OF CYTOKINE RESPONSES IN FIV

这项提案的目标是全面描述 细胞因子mRNA和蛋白在鸡淋巴组织中的表达 感染FIV的猫并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：不适当的细胞因子产生 导致对FIV的无效免疫反应，病理- FIV的起源和感染猫对FIV的易感性 机会性感染。细胞因子的调节可能有助于 研究治疗性干预和疫苗的有效性 策略，以及FIV的发病机制。具体目标1： 检测FIV感染对细胞因子mRNA和DNA的影响 猫多种淋巴组织中蛋白质的产生 间隔室将通过QC-RT-PCR进行评估以量化组织 通过原位杂交定位细胞因子mRNA的产生 在组织结构中，通过免疫细胞化学， ELISPOT和流式细胞仪检测细胞因子蛋白 在特定的细胞表型内产生。具体目标2： 测定IL-2、IL-4、IL-10、IL-12、肿瘤坏死因子-α及 干扰素-γ在FIV、抗IL-2、抗IL-4、 抗IL10、抗IL12p40、抗肿瘤坏死因子-α或抗-干扰素-γ 单抗和多克隆抗体将用于治疗 感染FIV病毒的猫。细胞因子抑制对血管内皮细胞生长的影响 病毒复制、免疫反应和疾病进展将是 下定决心。具体目标3：确定一个强大的细胞是否 对FIV的免疫是保护性的，SPF猫将接种活疫苗 重组生物疫苗载体；FIVenv和/或FIVgag 表达单核细胞增多性李斯特菌。然后，猫将受到挑战 同源和异种FIV与免疫应答 与保护性或非保护性相关的细胞因子谱 将对保护性免疫进行评估。意义：慢病毒- 相关免疫缺陷是一个不断扩大的全球问题 然而，影响人类和猫的免疫病理机制是 不完全了解，目前的治疗方法有限。 了解细胞因子在发病机制中的作用， 治疗和接种疫苗将对FIV感染产生直接影响 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.