Terahara Kazutaka, Iwabuchi Ryutaro, Hosokawa Masahito, Nishikawa Yohei, Takeyama Haruko, Takahashi Yoshimasa, Tsunetsugu-Yokota Yasuko
Department of Immunology, National Institute of Infectious Diseases, 1-23-1 Toyama, Shinjuku-ku, Tokyo, 162-8640, Japan.
Department of Life Science and Medical Bioscience, Waseda University, 2-2 Wakamatsu-cho, Shinjuku-ku, Tokyo, 162-8480, Japan.
BMC Res Notes. 2019 Apr 29;12(1):242. doi: 10.1186/s13104-019-4281-5.
Resting CD4 T cells are major reservoirs of latent HIV-1 infection, and may be formed during the early phase of the infection. Although CCR5-tropic (R5) HIV-1 is highly transmissible during the early phase, newly infected individuals have usually been exposed to a mixture of R5 and CXCR4-tropic (X4) viruses, and X4 viral DNA is also detectable in the host. Our aim was to identify which subsets of resting CD4 T cells contribute to forming the latent reservoir in the presence of both X4 and R5 viruses.
Primary resting CD4 naïve T (T) cells, CCR5 memory T (T) cells, and CCR5 T cells isolated by flow cytometry were infected simultaneously with X4 and R5 HIV-1, which harbored different reporter genes, and were cultured in the resting condition. Flow cytometry at 3 days post-infection demonstrated that X4 HIV-1 cells were present in all three subsets of cells, whereas R5 HIV-1 cells were present preferentially in CCR5 T cells, but not in T cells. Following CD3/CD28-mediated activation at 3 days post-infection, numbers of R5 HIV-1 cells and X4 HIV-1 cells increased significantly only in the CCR5 T subset, suggesting that it provides a major reservoir of replication-competent, latently infected viruses.
静息CD4 T细胞是潜伏性HIV-1感染的主要储存库,可能在感染早期形成。尽管CCR5嗜性(R5)HIV-1在感染早期具有高度传染性,但新感染个体通常接触过R5和CXCR4嗜性(X4)病毒的混合物,并且在宿主中也可检测到X4病毒DNA。我们的目的是确定在存在X4和R5病毒的情况下,静息CD4 T细胞的哪些亚群有助于形成潜伏储存库。
通过流式细胞术分离的原代静息CD4初始T(T)细胞、CCR5记忆T(T)细胞和CCR5 T细胞同时感染携带不同报告基因的X4和R5 HIV-1,并在静息条件下培养。感染后3天的流式细胞术显示,X4 HIV-1细胞存在于所有三个细胞亚群中,而R5 HIV-1细胞优先存在于CCR5 T细胞中,而不存在于T细胞中。在感染后3天进行CD3/CD28介导的激活后,仅在CCR5 T亚群中,R5 HIV-1细胞和X4 HIV-1细胞的数量显著增加,这表明它是具有复制能力的潜伏感染病毒的主要储存库。