猕猴艾滋病模型EKM基因治疗效果观察

目的明确体外导入EKM基因的实验猕猴感染SHIV-KB9后，病毒复制和免疫反应情况，为基因治疗AIDS提供实验依据。方法用血清学方法筛选出无SIV、STLV、SRV／D和B病毒感染的猕猴7只，实验当天回输自体储备血75ml，同时采集猴外周血75m1进行CD4＋T细胞分离培养，并导入EKM基因，培养结束后收获细胞，静脉回输猴体后再进行SHIV-KB9感染。采用流式细胞术、血常规检测、病毒栽量检测和基因拷贝数检测的方法确定导入基因后的实验猕猴感染sHIV．KB9后体内病毒复制和免疫损伤情况。结果导入基因和回输细胞的实验猕猴其血浆病毒载量峰值都与已报道的推迟14d出现，比模型对照组推迟11d出现，血浆病毒载量峰值均比模型对照组峰值低。流式细胞术结果表明其CD4＋T细胞计数也稳定在一定水平，CD4／CD8比值未出现明显倒置。实验组猕猴在试验结束时均未发展为猴AIDS。结论自体回输导入EKM基因的CD4＋T细胞的猕猴AIDS模型其血浆病毒载量峰值与模型对照组峰值比较有迟后作用，且低于模型对照组峰值，与回输细胞的实验对照组没有明显区别，其更确定的效果还有待进一步的研究。

Observation on EKM Gene Therapy in AIDS Rhesus Monkey Model

Objective To evaluate the effect of EKM gene therapy in SHIV animal models. Methods Seven rhesus monkeys were selected and screened by serological test to ensure that they were free of SIV, SRV, STLV and BV infection. PBMC were collected for isolating CD4＋T and proliferate ex vivo and transduce EKM gene. When cell culture was over, the cells（transduced or not transduced） was harvested and re-infused the cells into rhesus macaques. Finally, all rhesus monkeys were inoculated with SHIV-KB9. Blood samples were collected at different time according to the time schedule, and used for blood composition anaysis, and prepare both plasma and PBMC for viral RNA detection by RT-PCR and FACS / EKM gene copy assay, respectively. Results The viral load peak was put off 11 days contrast to model group and about 2 weeks late according to documents, the level of plasma viremia was lower than model group, CD4＋T cell count and CD4/CD8 ratio were stable. All animals did not involved in AIDS at the end of this experiment. Conclusion Re-infusing the transduced cells into monkeys can delay the viral load peak, and the level of plasma viremia was lower than model group, which is no obvious difference between control group and experimental group, the exact effect of EKM will be researched further.