Expression and functional analysis of cellular anti-HIV factor among HIV-infected Long-Term Nonprogressor

1、寻找hiv感染的长期不进展者我们在日本大阪国立医院和国际医疗中心的2100多名住院患者中检索了CD4+ t细胞计数维持在500以上的近6年的患者。结果发现15例患者。2、apobec3g表达我们接种了抗APOBEC3G抗体，通过FI检测成功显示APOBEC3G在细胞内高表达。而在PBMC中，这种表达水平在每个批次中都有上升和下降的趋势。因此，我们构建了持续表达APOBEC3G蛋白的标准细胞系，该细胞系可用于APOBEC3G的规范化。通常，非贴壁细胞用于HIV感染试验。然而，APOBEC3G的持续表达和维持是困难的。然后，我们在贴壁细胞系293T细胞的基础上建立细胞系。在克隆A3G-C1和A3G-C4中，vif缺陷的HIV不能感染。APOBEC3G在这些细胞中的高表达和功能性。通过这些细胞系，可以定量测定APOBEC3G，并测量长期非进展者和慢进展者APOBEC3G的表达。

1,Search of HIV-infected Long Term NonprogressorsWe searched the patients who maintained CD4+-T Cell count more than 500 for recent six years among more than 2100 patients who hospitalized to Osaka National hospital and International Medical Center of Japan. As a result, 15 patients were found.2,Expression of APOBEC3GWe received anti-APOBEC3G antibody and it successfully showed the high expression of intracellular APOBEC3G through the FI assay. However, this expression level in PBMC goes up and down in each lot. Thus, we constructed the standard cell line which expresses the APOBEC3G protein constantly, and this cell line was useful for the normalization of APOBEC3G. Generally, non-Adherent cells were used for the HIV infection assay. However, it is difficult to make and maintain the appropriate APOBEC3G expression constantly. Then, we establish the cell line which is based on the adherent cell line, 293T cells. Among the clone A3G-C1 and A3G-C4, vif-defected HIV could not infect with. APOBEC3G expression in these cells was high and functional. By these cell line, quantification of APOBEC3G is possible and the measurement of APOBEC3G expression among Long Term Nonprogressors and slow progressors.