猴免疫缺陷病毒(SIV)在艾滋病模型恒河猴组织中的分布和载量测定

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目的研究当艾滋病恒河猴模型的血浆病毒载量处于低水平或阴性时,猴免疫缺陷病毒(simian immunodeficiency viruses,SIV)在宿主组织中的分布情况。方法SIVmac251感染恒河猴10只,定期检测其血浆载量,感染病毒平均高峰时间第14天时,活检取淋巴结。选取感染18个月后病毒载量最低水平和阴性的2只艾滋病猴(SAIDS),经安死术后取淋巴结、脾、肝、肺、肾、脑等组织,用原位杂交和实时荧光定量PCR的方法检测病毒在组织中的分布和组织中的病毒载量。结果感染后14d,10只猴血浆病毒载量达到107copies/mL,淋巴结组织病毒载量为105~108copies/g,原位杂交方法在腹股沟淋巴结中检测到强阳性斑点。感染后第18个月的2只猴,血浆病毒载量下降并维持不高于102copies/mL水平或阴性,但组织分布不尽相同,在肠系膜淋巴结、肾上腺、海马回、空肠、脾脏等组织中检测到105~106copies/g的病毒载量,于一只猴的脑积液中检测到103copies/mL的病毒载量。用原位杂交的方法在肠系膜淋巴结和空肠中检测到强阳性斑点,其它组织中未检测到阳性斑点。结论实验证实SAIDS猴在血浆病毒载量低甚至阴性时,病毒在不同组织中仍有分布,有些组织中甚至出现高病毒载量,提示在制备SIV/SAIDS模型中,尤其在药物筛选和疫苗评价时,应考虑组织病毒载量指标的测定和药物、疫苗对组织病毒的治疗清除作用的评价。 Objective To investigate the distribution of simian immunodeficiency viruses (SIV) in host tissues when the plasma viral load of HIV-infected rhesus monkeys is low or negative. Methods 10 Rhesus macaques were infected with SIVmac251 and their plasma levels were measured regularly. The average infection time was 14 days. The lymph nodes were obtained by biopsy. Two AIDS-infected monkeys (SAIDS) with the lowest viral load and the lowest virulence after 18 months of infection were selected and lymph node, spleen, liver, lung, kidney and brain tissues were collected after antineoplastic surgery. The hybridization and real-time fluorescence quantitative The PCR method detects the distribution of the virus in the tissue and the viral load in the tissue. Results On the 14th day after infection, the plasma viral loads of 10 monkeys reached 107copies / mL and the viral load of lymph node tissue was 105 ~ 108copies / g. Strong positive spots were detected in inguinal lymph nodes by in situ hybridization. The two monkeys infected at 18 months postinfection showed a decrease in plasma viral load of no more than 102copies / mL or negative, but the tissue distribution was not the same. In the mesenteric lymph nodes, adrenal gland, hippocampus, jejunum, spleen and other tissues A viral load of 105 to 106 copies / g was detected and a viral load of 103 copies / mL was detected in the brain effusion of a monkey. Strong positive spots were detected in mesenteric lymph nodes and jejunum by in situ hybridization and no positive spots were detected in other tissues. Conclusion Experiments confirmed that the SAIVS virus in the plasma when the virus load is low or even negative, the virus is still distributed in different tissues, some tissues even high viral load, suggesting that in the preparation of SIV / SAIDS model, especially in drug screening and vaccine evaluation Should consider the determination of viral load indicators of the organization and evaluation of the therapeutic clearance of the virus by the drugs and vaccines.
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