艾滋病CD4+T细胞焦亡影响ITAMs/ITIMs炎症调控及中医补肾解毒法的干预作用

Pyroptosis was proved to drive the HIV infection induced CD4+T cell death. It is an important cause of continuous and exceeding immune activation and inflammation in AIDS. The ITAMs/ITIMs parthway was the main modulator for the inflammation and immune reaction which ensure the host does not be injured. This raised the question that if the pyroptosis can directly induce the abnormal regulation of ITAMs/ITIMs parthway? If the Kidney norishing and detoxication therapy realized its effect through restore the ITAMs/ITIMs parthway? This project planed to use the human lymphoid aggregate culture(HLAC) and simian AIDS model as the in vitro and in vivo model for addressing the affection of the HIV induced pyroptosis to the ITAMs/ITIMs parthway, and then identify the target receptors which include the ITAMs/ITIMs affected by the pyroptosis. Additionally, this project planed to block the ITAMs/ITIMs related key pathway and receptors with antibodies and siRNA for observing if the blocking capable of relieving the continuous and exceeding immune activation and inflammation in AIDS. This will provide new idea for AIDS treatment. This project also plan to observe the effect of the Kidney norishing and detoxication therapy to HIV induced pyroptosis affected key ITAMs/ITIMs parthway and related receptors. This will deeply reveal the mechanism of the Chinese medicine treatment of AIDS.

“细胞焦亡”被证实为HIV感染导致CD4+T细胞死亡的主要方式，是艾滋病持续过度的炎症和免疫激活的重要原因。机体主要通过ITAMs/ITIMs通路调控炎症和免疫反应使其不至于过度而伤害自身。那么HIV感染引起的“细胞焦亡”是否可直接导致ITAMs/ITIMs通路调控异常？中医补肾解毒法是否通过修复ITAMs/ITIMs通路而实现其效果？本项目拟以人淋巴细胞聚集培养（HLACs）和猴艾滋病模型为主要体内外模型，阐明HIV感染导致的细胞焦亡对ITAMs/ITIMs通路的影响、鉴定出具体受影响的包含ITAMs/ITIMs的受体，并采用抗体、siRNA等手段阻断焦亡影响的ITAMs/ITIMs关键通路及相关受体观察是否可减轻免疫激活和炎症程度，为艾滋病治疗提示新思路。观察补肾解毒法对艾滋病细胞焦亡导致ITAMs/ITIMs关键通路及相关受体通路紊乱的影响，深刻揭示中医治疗艾滋病的疗效机制。

“细胞焦亡”是艾滋病持续过度的炎症和免疫激活的重要原因。机体主要通过ITAMs/ITIMs通路调控炎症和免疫反应使其不至于过度而伤害自身。那么HIV感染引起的“细胞焦亡”是否可直接导致ITAMs/ITIMs通路调控异常？中医补肾解毒法是否通过修复ITAMs/ITIMs通路而实现其效果？本项目以人淋巴细胞聚集培养（HLACs）和猴艾滋病模型为主要体内外模型，阐明HIV感染导致的细胞焦亡对ITAMs/ITIMs通路的影响，观察补肾解毒法对艾滋病细胞焦亡导致ITAMs/ITIMs关键通路及相关受体通路紊乱的影响。体外实验结果显示，HIV感染导致的细胞焦亡可使ITAMs/ITIMs炎症调控通路中的SHP-1mRNA和蛋白表达显著升高，同时IL-6、IL-18、TNF-α、TRAF-4mRNA等等炎症相关因子表达显著升高；对SHP-1进行的基因沉默实验显示，SHP-1mRNA沉默后，IL-6、IL-18、TNF-α、TRAF-4mRNA均显著下降；说明SHP-1是HIV感染后炎症调控通路中最明显的细胞因子。补肾解毒法体外作用后，可显著降低SHP-1、IL-6、IL-18、TNF-α、TRAF-4mRNA，且可明显抑制HIV病毒量；说明补肾解毒法可能通过调控SHP-1而减轻炎症；而猴艾滋病动物模型实验显示，SIV感染猴外周血PBMC及浅表淋巴结中SHP-1mRNA显著升高，且IL-6，IL-18，IL-1β，TNF-α，IL-33等炎症因子mRNA显著升高，补肾解毒法高剂量组可使PBMC及浅表淋巴结SHP-1及IL-6，IL-18，IL-1β，TNF-α，IL-33等炎症因子显著降低，说明补肾解毒法在体内也可修复炎症调控通路从而减轻SIV感染导致的免疫激活和炎症。本项目证实了HIV感染导致细胞焦亡可以显著影响炎症调控通路的假说，且补肾解毒法可以修复已经改变的炎症调控通路，为中医治疗艾滋病的具体机制提供了实验支持。