Appiah-Kubi Joyce, Yuzhe Yuan, Kuwata Takeo, Terasawa Hiromi, Nyame Perpetual, Amesimeku Wright Ofotsu, Hossain Md Jakir, Matsushita Shuzo, Sawa Tomohiro, Maeda Yosuke, Harada Shinji, Yusa Keisuke, Monde Kazuaki
Department of Microbiology, Faculty of Life Sciences, Kumamoto University, Kumamoto, 860-8556, Japan.
Department of Virology, Noguchi Memorial Institute for Medical Research, University of Ghana, Accra, Ghana.
Sci Rep. 2025 Jul 21;15(1):26413. doi: 10.1038/s41598-025-12653-9.
For the entry of R5 human immunodeficiency virus type 1 (HIV-1) into target cells, CCR5 functions as a coreceptor following the binding of the envelope glycoprotein gp120 to the receptor CD4. While CD4 is known to be strongly downregulated after infection, the fate of CCR5 post-infection remains unclear. We investigated the surface expression of CCR5 on PM1/CCR5 cells following infection with HIV-1. Flow cytometry using anti-CCR5 MAb (T21/8 and 2D7) revealed that CCR5 was not downregulated on the surface of infected cells. Notably, CCR5 was found to be coassembled with HIV-1 Gag at the viral budding sites. A specific subset of CCR5, designated as CCR5, which is recognized by T21/8 but not 2D7, was significantly colocalized with gp120 on the cell surface. Virions incorporating CCR5 were immunoprecipitated using T21/8 MAb but not with 2D7 Mab, indicating that CCR5 was incorporated into progeny virions. The efficiency of CCR5 incorporation into virions was positively correlated with the level of CCR5 expression on the cell surface. Moreover, incorporation efficiency was approximately 83-fold higher in CD4 T cells than in adherent CCR5 cells. Furthermore, the incorporation of CCR5 into HIV-1 virions resulted in a 25% decrease in viral infectivity. These findings suggest that CCR5 may have a detrimental effect during the late stage of the HIV-1 replication cycle.
对于R5型人类免疫缺陷病毒1型（HIV-1）进入靶细胞而言，在包膜糖蛋白gp120与受体CD4结合后，CCR5作为共受体发挥作用。虽然已知CD4在感染后会强烈下调，但感染后CCR5的命运仍不清楚。我们研究了HIV-1感染后PM1/CCR5细胞表面CCR5的表达情况。使用抗CCR5单克隆抗体（T21/8和2D7）进行的流式细胞术显示，感染细胞表面的CCR5没有下调。值得注意的是，发现CCR5在病毒出芽位点与HIV-1 Gag共同组装。CCR5的一个特定亚群，称为CCR5，可被T21/8识别但不能被2D7识别，在细胞表面与gp120显著共定位。使用T21/8单克隆抗体而非2D7单克隆抗体免疫沉淀掺入CCR5的病毒颗粒，表明CCR5被掺入子代病毒颗粒中。CCR5掺入病毒颗粒的效率与细胞表面CCR5的表达水平呈正相关。此外，CD4 T细胞中的掺入效率比贴壁CCR5细胞高约83倍。此外，CCR5掺入HIV-1病毒颗粒导致病毒感染性降低25%。这些发现表明，CCR5可能在HIV-1复制周期的后期产生有害影响。
