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目的为建立SHIV艾滋病动物模型提供毒力较强的病毒株,将新合成的SHIV XJ02170病毒适应猴体,并增强其毒力。方法实验前采集猴血清并进行血清学检查和PCR检测。选出13只无SIV,STLV1,SRV D和B病毒感染的猴。第一批实验,将SHIV前病毒DNA质粒经肌肉注射到猴体内,每只500μg;SHIV病毒液,经静脉注射到猴体内,每只2ml。病毒质粒和病毒液各接种2只猴。当第一批猴体检出病毒后,10ml感染猴的全血,抗凝后静脉注射到第二批猴体内,当第二批猴检出病毒后再将10ml感染猴的全血静脉注射到第三批猴体内,连续传代4次。每批实验均定期采集血液标本,分别用肝素和EDTA抗凝,进行病毒分离;病毒基因PCR检测;CD4,CD8测定;病毒抗体检测。结果SHIV XJ02170病毒和SHIV XJ02170前病毒DNA质粒在猴体内的传代中均能分离出病毒;从传代猴的血浆和外周血单核细胞(PBMC)中检出了病毒DNA和RNA基因;CD4,CD8测定结果显示有暂时性倒置现象,后变为正常倒置与正常交替出现;在传代的猴血清中检测出特异性HIV病毒抗体。结论SHIV XJ02170病毒与SHIV XJ02170前病毒DNA质粒,均能在恒河猴体内复制。
OBJECTIVE: To provide a virulent strain for the establishment of an animal model of SHIV AIDS. The newly synthesized SHIV XJ02170 virus was adapted to the monkeys and its toxicity was enhanced. Methods Monkeys were collected before experiment and serological test and PCR test. Thirteen monkeys infected with SIV, STLV1, SRV D and B virus were selected. In the first experiments, the SHIV proviral DNA plasmid was intramuscularly injected into the monkeys, each 500μg; SHIV virus solution, intravenous injection into the monkey, each 2ml. Viral plasmids and virus solution were inoculated with 2 monkeys. When the first batch of monkeys detected the virus, the whole blood of 10ml infected monkeys were injected intravenously into the second batch of monkeys after anticoagulation. When the second batch of monkeys detected the virus, 10ml of whole blood was injected into the Three batches of monkeys, continuous passage 4 times. Blood samples were collected regularly in each batch of experiments, anticoagulated with heparin and EDTA respectively for virus isolation; PCR detection of viral genes; CD4, CD8 assay; and detection of virus antibodies. Results The virus was isolated from the SHIV XJ02170 and SHIV XJ02170 proviral DNA plasmids in passaged monkeys. The viral DNA and RNA genes were detected from the plasma and peripheral blood mononuclear cells (PBMCs) The results showed that there is a temporary inversion phenomenon, after the change into a normal alternating with the normal alternating; detected in the passage of the monkey serum specific HIV antibody. Conclusion Both SHIV XJ02170 and SHIV XJ02170 proviral DNA plasmids can replicate in rhesus monkeys.