Department of Medical Biotechnologies, University of Siena, Siena, Italy.
VisMederi srl, Siena, Italy.
J Gen Virol. 2021 Jan;102(1). doi: 10.1099/jgv.0.001499.
Recent studies have suggested that the CCR5 antagonist maraviroc (MVC) may exert an HIV-1 latency reversal effect. This study aimed at defining MVC-mediated induction of HIV-1 in three cell line latency models and in CD4 T cells from six patients with suppressed viraemia. HIV-1 induction was evaluated in TZM-bl cells by measuring HIV-1 LTR-driven luciferase expression, and in ACH-2 and U1 latently infected cell lines by measuring cell-free (CFR) and cell-associated (CAR) HIV-1 RNA by qPCR. NF-B p65 was quantified in nuclear extracts by immunodetection. In CD4 T cells, CAR, CFR and cell-associated DNA (CAD) were quantified at baseline and 1-7-14 days post-induction (T1, T7, T14). At T7 and T14, the infectivity of the CD4 T cells co-cultured with MOLT-4/CCR5 target cells was evaluated in the TZM-bl assay (TZA). Results were expressed as fold activation (FA) with respect to untreated cells. No LTR activation was observed in TZM-bl cells at any MVC concentration. NF-B activation was only modestly upregulated (1.6±0.4) in TZM-bl cells with 5 µM MVC. Significant FA of HIV-1 expression was only detected at 80 µM MVC, namely on HIV-1 CFR in U1 (3.1±0.9; =0.034) and ACH-2 cells (3.9±1.4; =0.037). CFR was only weakly stimulated at 20 µM in ACH-2 (1.7±1.0 FA) cells and at 5 µM in U1 cells (1.9±0.5 FA). Although no consistent pattern of MVC-mediated activation was observed in experiments, substantial FA values were detected sparsely on individual samples with different parameters. Notably, in one sample, MVC stimulated all parameters at T7 (2.3±0.2 CAD, 6.8±3.7 CAR, 18.7±16.7 CFR, 7.3±0.2 TZA). In conclusion, MVC variably induces HIV-1 production in some cell line models not previously used to test its latency reversal potential. In CD4 T cells, MVC may exert patient-specific HIV-1 induction; however, clinically relevant patterns, if any, remain to be defined.
最近的研究表明，趋化因子受体 5 拮抗剂马拉维若（MVC）可能发挥 HIV-1 潜伏期逆转作用。本研究旨在定义 MVC 在三种细胞系潜伏期模型和六位病毒血症抑制患者的 CD4 T 细胞中诱导 HIV-1 的作用。通过测量 HIV-1 LTR 驱动的荧光素酶表达，在 TZM-bl 细胞中评估 HIV-1 的诱导，在 ACH-2 和 U1 潜伏感染细胞系中通过 qPCR 测量细胞外（CFR）和细胞相关（CAR）HIV-1 RNA。通过免疫检测在核提取物中定量 NF-B p65。在 CD4 T 细胞中，在诱导后 1-7-14 天（T1、T7、T14）时基线和 CAR、CFR 和细胞相关 DNA（CAD）进行定量。在 T7 和 T14，与 MOLT-4/CCR5 靶细胞共培养的 CD4 T 细胞的感染性在 TZM-bl 测定中进行评估（TZA）。结果以相对于未处理细胞的激活倍数（FA）表示。在任何 MVC 浓度下，TZM-bl 细胞均未观察到 LTR 激活。在 5 µM MVC 的 TZM-bl 细胞中，NF-B 激活仅适度上调（1.6±0.4）。仅在 80 µM MVC 时才检测到 HIV-1 表达的显著 FA，即在 U1（3.1±0.9；=0.034）和 ACH-2 细胞（3.9±1.4；=0.037）中。在 ACH-2 细胞（1.7±1.0 FA）中，在 20 µM 时仅弱刺激 CFR，在 U1 细胞中在 5 µM 时（1.9±0.5 FA）。尽管在实验中没有观察到 MVC 介导的激活的一致模式，但在不同参数的个别样本中稀疏地检测到大量 FA 值。值得注意的是，在一个样本中，MVC 在 T7 时刺激所有参数（2.3±0.2 CAD、6.8±3.7 CAR、18.7±16.7 CFR、7.3±0.2 TZA）。总之，MVC 可在以前未用于测试其潜伏期逆转潜力的一些细胞系模型中可变地诱导 HIV-1 产生。在 CD4 T 细胞中，MVC 可能发挥患者特异性 HIV-1 诱导作用；然而，如果存在任何临床相关模式，仍有待定义。
