基于小鼠白血病细胞株的HIV-1感染小鼠动物模型及抗HIV中药体内疗效研究

Research of in vivo efficacy of anti-HIV Chinese medical herbs, generally adopted non-human primate animal models, which exists less animal resources, high cost, small sample size, etc. Establishment of a HIV infected mouse model with low cost and high application value can offer an effective tool to the research on the anti-HIV effects of Chinese medical herbs. .In our preliminary work, hCD4/CCR5/Cyclin T1 were transfected by the lentiviral vector transfection to the mouse leukemia cell lines L615 and L1210, and the results found that there are high load of HIV-1 replication in the two cell lines..In this study, the murine leukemia virus induced-mouse leukemia cell lines L615 and L1210 were used, and then make the cell lines infected with the hCD4/CCR5/Cyclin T1 loaded-lentiviral vector. After the protein of human CD4 and CCR5 being expressed on the cell lines surface, the mouse leukemia cells will be humanized, and possess the characteristic which is necessary when HIV infect with the human host cells and virus replication factor. The animal model of leukemia mice was prepared by transplanted the mouse leukemia cells into the DBA/2 or BALB/C mouse with intravenous injection, and a HIV infected animal model of mouse was established. .And then, some effective traditional Chinese medicine by literature confirmed that could inhibit HIV replication, and the anti-HIV medical herbs discovered in our previous in vitro studies, were evaluated the in vivo anti-HIV efficacies, by using this HIV-1 infected mouse model. The HIV-1 infected mouse model established in our research has the advantages of abundant experimental animal resource, simple operation and low cost in the anti-HIV in vivo efficacy research of Chinese medicine herbs.

抗HIV中药的体内疗效研究，普遍采用非人灵长类动物模型，存在动物资源匮乏、成本高、样本少等缺点。建立一个低成本、高应用价值的鼠类HIV感染动物模型，有利于促进抗HIV中药的研究。.前期工作，我们应用慢病毒载体转染hCD4/CCR5/Cyclin T1于小鼠白血病细胞株L615和L1210后，实现了HIV-1的感染及其病毒复制。.本研究中，应用hCD4/CCR5/Cyclin T1慢病毒过表达系统，使白血病细胞株L615和L1210具备融合入胞的受体与辅助受体、病毒复制因子，建立HIV-1的宿主细胞；将此白血病细胞移植接种于DBA/2近交系鼠或BALB/C小鼠，制备成HIV易感小鼠动物模型。.然后，应用本研究的动物模型，对文献报道证实的多种抑制HIV中药、以及我们体外实验新发现的7种抗HIV有效中药进行体内疗效评价。本模型应用于抗HIV中药的体内疗效评估，具有成本低、效率高的特点。

抗HIV中药的体内疗效研究，普遍采用非人灵长类动物模型，存在动物资源匮乏、成本高、样本少等缺点。建立一个低成本、高应用价值的鼠类HIV-1感染动物模型，有利于促进抗HIV-1中药的在体疗效研究。.本研究中，我们应用慢病毒载体转染hCD4/CCR5/Cyclin T1于小鼠白血病细胞株L615和L1210后，使白血病细胞株L615和L1210具备融合入胞的受体/辅助受体和病毒复制因子，实现了HIV-1的感染及其病毒复制，建立HIV-1的宿主细胞；也就是说实现了HIV-1对鼠细胞的跨物种的感染。.将稳定表达hCD4/CCR5/Cyclin T1的小鼠白血病细胞移植接种于普通裸鼠尾静脉，检测动物血液中移植的白血病细胞比率以及鼠白血病细胞膜hCD4/CCR5的表达情况；HIV-1感染小鼠后第7、14、21、28天处死动物，检测血液中的病毒载量，结果证实验制备成HIV易感小鼠动物模型。.最后，应用我们建立的HIV小鼠模型，验证抗病毒药物的抑制病毒复制作用；同时，将文献报道的具有抗HIV作用的中药约4味、以及我们在体外实验证实具有抗HIV作用的6种中药，进行抗HIV-1药物动物体内疗效观察，结果显示有3个药物单体具有抗病毒作用，其作用机制尚在探索中。.简而言之，在研究期间，我们实现了跨物种感染HIV-1的鼠细胞系2个，成功建立了基于裸鼠的HIV-1感染动物模型，进行了5个中药（及单体物）的体内疗效观察，参加了国内学术会议3次，培养了3位硕士研究生，发表了国内论文3篇（2篇为核心期刊），标注SCI论文3篇。中药抗HIV-1的体疗效观察，因需明确该药物抗病毒的作用机制，论文尚待发表。