Okutomi Toshiki, Minakawa Satoko, Hirota Riku, Katagiri Koko, Morikawa Yuko
Graduate School of Infection Control Sciences, Kitasato University, Shirokane 5-9-1, Minato-ku, Tokyo 108-8641, Japan.
Department of Biosciences, School of Science, Kitasato University, Kitasato 1-15-1, Minami-ku, Sagamihara, Kanagawa 252-0373, Japan.
Viruses. 2020 Apr 8;12(4):417. doi: 10.3390/v12040417.
HIV reactivation from latency is induced by cytokines but also by cell contact with other cells. To better understand this, J1.1 cells, a latent HIV-1-infected Jurkat derivative, were cocultured with its parental Jurkat. J1.1 cells became p17MA-positive and produced a high level of HIV p24CA antigen, only when they were cocultured with stimulated Jurkat with cell-to-cell contact. In contrast, very little p24CA was produced when they were cocultured without cell contact. Similar results were obtained when latent ACH-2 and its parental A3.01 cells were cocultured. Confocal microscopy revealed that not only HIV-1 p17MA and gp120Env but also LFA-1, CD81, CD59, and TCR CD3 accumulated at the cell contact site, suggesting formation of the virological synapse-like structure. LFA-1-ICAM-1 interaction was involved in the cell-to-cell contact. When J1.1 was cocultured with TCR-deficient Jurkat, the p17MA-positive rate was significantly lower, although the cell-to-cell contact was not impaired. Quantitative proteomics identified 54 membrane molecules, one of which was MHC class I, that accumulated at the cell contact site. Reactivation from latency was also influenced by the presence of stromal cells. Our study indicated that latent HIV-1 in J1.1/ACH-2 cells was efficiently reactivated by cell-to-cell contact with stimulated parental cells, accompanying the virological synapse-like structure.
潜伏的HIV从潜伏期重新激活是由细胞因子诱导的，但也可由与其他细胞的细胞接触诱导。为了更好地理解这一点，将J1.1细胞（一种潜伏感染HIV-1的Jurkat衍生物）与其亲本Jurkat细胞共培养。只有当J1.1细胞与经刺激的Jurkat细胞进行细胞间接触共培养时，它们才会变成p17MA阳性并产生高水平的HIV p24CA抗原。相反，在没有细胞接触的情况下共培养时，产生的p24CA很少。将潜伏的ACH-2细胞与其亲本A3.01细胞共培养时也得到了类似的结果。共聚焦显微镜显示，不仅HIV-1 p17MA和gp120Env，而且LFA-1、CD81、CD59和TCR CD3都在细胞接触部位聚集，提示形成了病毒学突触样结构。LFA-1-ICAM-1相互作用参与了细胞间接触。当J1.1与缺乏TCR的Jurkat细胞共培养时，尽管细胞间接触未受损，但p17MA阳性率显著降低。定量蛋白质组学鉴定出54种在细胞接触部位聚集的膜分子，其中之一是MHC I类分子。潜伏期的重新激活也受基质细胞存在的影响。我们的研究表明，J1.1/ACH-2细胞中潜伏的HIV-1通过与经刺激的亲本细胞的细胞间接触而有效重新激活，并伴有病毒学突触样结构。
