Maikoo Shreyal, Palstra Robert-Jan, Dong Krista L, Mahmoudi Tokameh, Ndung'u Thumbi, Madlala Paradise
HIV Pathogenesis Programme, The Doris Duke Medical Research Institute, Nelson R. Mandela School of Medicine, University of KwaZulu-Natal, Durban, South Africa.
School of Laboratory Medicine and Medical Sciences, University of KwaZulu-Natal, Durban, South Africa.
J Virus Erad. 2024 Dec 13;10(4):100575. doi: 10.1016/j.jve.2024.100575. eCollection 2024 Dec.
Sub-Saharan Africa accounts for almost 70 % of people living with HIV (PLWH) worldwide, with the greatest numbers centred in South Africa where 98 % of infections are caused by subtype C (HIV-1C). However, HIV-1 subtype B (HIV-1B), prevalent in Europe and North America, has been the focus of most cure research and testing despite making up only 12 % of HIV-1 infections globally. Development of latency models for non-subtype B viruses is a necessary step to address this disproportionate focus. Furthermore, the impact of genetic variation between viral subtypes, specifically within the long terminal repeat (LTR) element of the viral transcriptional promoter on latency reversal, remains unclear. To address this scientific gap, we constructed a minimal genome retroviral vector expressing HIV-1C consensus transactivator of transcription protein (Tat) and green fluorescent protein (GFP) under the control of either HIV-1C consensus LTR (C731CC) or the transmitted/founder (T/F) LTRs derived from PLWH (C731CC), produced corresponding LTR pseudotyped viruses using a vesicular stomatitis virus (VSV-G) pseudotyped Envelope vector and the pCMVΔR8.91 packaging vector containing HIV-1 accessory and genes. Viruses produced in this way were used to infect Jurkat E6 and primary CD4 T cells . By enriching for latently infected cells, and treating them with different latency reversing agents, we developed an HIV-1C latency model that demonstrated that the HIV-1C consensus LTR has lower reactivation potential compared to its HIV-1B counterpart. Furthermore, HIV-1C T/F LTR pseudotyped proviral genetic variants exhibited a heterogenous reactivation response which was modulated by host cell (genetic) variation. Our data suggests that genetic variation both within and between HIV-1 subtypes influences latency reversal. Future studies should investigate the specific role of variation in host cellular environment on reactivation differences.
撒哈拉以南非洲地区的人类免疫缺陷病毒（HIV）感染者占全球HIV感染者总数的近70%，其中绝大多数集中在南非，该国98%的感染是由C型（HIV-1C）亚型引起的。然而，在欧洲和北美流行的HIV-1 B亚型（HIV-1B），尽管仅占全球HIV-1感染的12%，却一直是大多数治愈研究和测试的重点。开发针对非B亚型病毒的潜伏模型是解决这种关注失衡问题的必要步骤。此外，病毒亚型之间的基因变异，特别是病毒转录启动子的长末端重复序列（LTR）元件内的变异对潜伏激活的影响仍不清楚。为了填补这一科学空白，我们构建了一种最小基因组逆转录病毒载体，该载体在HIV-1C共有LTR（C731CC）或来自HIV感染者的传播/奠基者（T/F）LTR的控制下，表达HIV-1C共有转录激活蛋白（Tat）和绿色荧光蛋白（GFP），使用水泡性口炎病毒（VSV-G）假型包膜载体和含有HIV-1辅助基因的pCMVΔR8.91包装载体产生相应的LTR假型病毒。以这种方式产生的病毒用于感染Jurkat E6细胞和原代CD4 T细胞。通过富集潜伏感染细胞并用不同的潜伏激活剂处理它们，我们建立了一个HIV-1C潜伏模型，该模型表明HIV-1C共有LTR与其HIV-1B对应物相比具有较低的激活潜力。此外，HIV-1C T/F LTR假型前病毒基因变体表现出异质性的激活反应，该反应受到宿主细胞（基因）变异的调节。我们的数据表明，HIV-1亚型内部和之间的基因变异会影响潜伏激活。未来的研究应调查宿主细胞环境变异对激活差异的具体作用。
