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

1、寻找HIV感染的长期无进展患者我们在大阪国立医院和日本国际医学中心住院的2100多名患者中搜索了近6年来CD4-T细胞计数保持在500以上的患者。结果发现15例APOBEC3G2、APOBEC3GG的表达我们接受了抗APOBEC3G的抗体，通过FI检测，成功地显示了APOBEC3G在细胞内的高表达。然而，这种在PBMC中的表达水平在每一批中都是上升和下降的。因此，我们构建了持续表达APOBEC3G蛋白的标准细胞系，为APOBEC3G的正常化奠定了基础。通常，非贴壁细胞用于HIV感染的检测。然而，要不断地制造和维持适当的APOBEC3G表达是困难的。在此基础上，建立了293T细胞贴壁细胞系。在克隆A3G-C1和A3G-C4中，病毒缺陷的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，APOBEC3G的定量以及APOBEC3G在长期无进展者和慢进展者中的表达的测量成为可能。

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.