Toll-Like Receptors in Bacterial Antigen Induced HIV-LTR

在HIV感染患者中，血浆HIV负荷增加伴随着机会性感染，包括分枝杆菌和衣原体感染，表明细胞激活和病毒活跃复制。先天免疫系统和Toll样受体(TLRs)在机会性感染期间HIV复制中的作用尚不清楚。我已经证明，内毒素通过IL-1R信号分子通过先天免疫系统受体TLR4激活HIV-Long末端重复序列(LTR)。在初步实验中，我观察到TLR2转导结核分枝杆菌抗原可溶性因子(STF)反应来激活HIV-LTR，并且TLR4(内毒素)和TLR2(STF)配体诱导HIV-LTR的协同激活。TLR2介导的STF激活HIV-LTR的分子机制以及TLR2和TLR4配体协同激活HIV-LTR的机制尚不清楚。此外，尚不清楚衣原体效应分子热休克蛋白60(CHSP60)是否被TLR4或TLR2识别。这项建议的总体目标是确定先天免疫受体的作用，以及在分枝杆菌和衣原体抗原诱导的HIV-1-LTR激活和复制中涉及的分子信号机制，以及TLR2和TLR4配体协同激活HIV-LTR的分子机制。假设：我假设通过TLRs(TLR2用于STF，TLR4用于LPS和cHSP60)激活天然免疫系统，通过IL-1R信号分子介导细菌和分枝杆菌抗原诱导的NF-kB和HIV-LTR的激活。我还假设TLR4和TLR2配体协同诱导HIV-LTR反式激活。因此，TLRs可能在结核分枝杆菌和衣原体感染等机会性感染与艾滋病毒复制增加之间提供潜在的分子联系。其具体目的是：(1)评估TLR2和TLR4的作用，并确定TLRs在可溶性结核因子(STF)和衣原体HSP60(CHSP60)诱导的巨噬细胞HIV-LTR激活中的功能协同和协同作用。(2)-探讨TLR20和TLR4诱导的HIV-LTR反式激活和HIV复制的分子信号机制。(2A)-确定IL-1信号分子Tollip、MyD88和NF-kB激活在TLR2介导的HIV-LTR激活中的作用。(2B)-研究PI3Kinase、rac1和Akt在TLR2和TLR4诱导的HIV-LTR激活和HIV复制中的作用。(2C)-研究TLR2和TLR4配体协同激活HIV-LTR的分子机制。意义：更好地理解先天免疫受体的作用，如TLRs，以及微生物抗原诱导的HIV-LTR反式激活和HIV复制所涉及的分子信号机制，可能为在分枝杆菌和衣原体感染期间更好地控制HIV复制提供新的靶点。

In HIV infected patients, plasma HIV load increases with concomitant opportunistic infections, including Mycobacterial and Chlamydial infections suggesting cell activation and active virus replication. The role of the innate immune system and Toll-like Receptors (TLRs) in HIV replication during opportunistic infections are unknown. I have shown that LPS activates HIV-long terminal repeat (LTR) through innate immune system receptor TLR4 via IL-1R signaling molecules. In preliminary experiments, I observed that TLR2 transduces mycobacterial antigen Soluble Tuberculosis Factor (STF) response to activate HIV-LTR, and that TLR4 (LPS) and TLR2 (STF) ligands induce synergistic activation of HIV-LTR. The molecular mechanisms involved in TLR2-mediated STF activation of HIV-LTR and the mechanisms of the synergy observed between TLR2 and TLR4 ligands for HIV-LTR transactivation are unknown. Furthermore, it is unknown whether Chlamydial effector molecule, Heat Shock Protein 60 (cHSP60) is recognized by TLR4 or TLR2. The overall goals of this proposal are to determine the role of the innate immune receptors, TLRs, and the molecular signaling mechanisms involved in mycobacterial and chlamydial antigen-induced HIV-1-LTR activation and replication, as well as the molecular mechanisms involved in the synergistic activation of HIV-LTR by TLR2 and TLR4 ligands. The hypothesis: I hypothesize that activation of the innate immune system through TLRs (TLR2 for STF, and TLR4 for LPS and cHSP60) mediate the bacterial and mycobacterial antigen-induced activation of NF-kB and HIV-LTR via IL-1R signaling molecules. I also hypothesize that TLR4 and TLR2 ligands synergistically induce HIV- LTR transactivation. Thus, TLRs may provide a potential molecular link between opportunistic infections such as Mycobacterial tuberculosis and Chlamydial infections and increased HIV replication. The specific aims are: (1)-To assess the role of TLR2 and TLR4, and to determine the functional cooperation and synergy of TLRs in Soluble Tuberculosis Factor (STF)-, and chlamydia HSP60 (cHSP60)-induced HIV-LTR activation in macrophages. (2)-To investigate the molecular signaling mechanisms involved in TLR20 and TLR4-induced HIV-LTR transactivation and HIV replication. (2a)-To determine the role of the IL-1 signaling molecules, Tollip, MyD88 and NF-kB activation in TLR2-mediated HIV-LTR activation. (2b)-To study the role of PI3 Kinase, Rac1 and Akt in TLR 2 and TLR4-induced activation of HIV- LTR and HIV replication. (2c)-To investigate the molecular mechanism of the synergy between TLR2 and TLR4 ligands forHIV-LTR activation. Significance: Improved understanding of the role of innate immune receptors, such as TLRs and the molecular signaling mechanisms involved in microbial antigen induced HIV-LTR transactivation and HIV replication may provide new targets to better control HIV replication during mycobacterial and chlamydial infections.