Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA.
Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, Massachusetts, USA
J Virol. 2019 Jul 30;93(16). doi: 10.1128/JVI.00731-19. Print 2019 Aug 15.
Recently, we and others have shown that natural killer (NK) cells exhibit memory-like recall responses against cytomegalovirus (CMV) and human immunodeficiency/virus simian immunodeficiency virus (HIV/SIV) infections. Although the mechanism(s) have not been fully delineated, several groups have shown that the activating receptor NKG2C is elevated on NK cells in the context of rhesus CMV (rhCMV) or human CMV (hCMV) infections. CD94, which heterodimerizes with NKG2C is also linked to adaptive NK cell responses. Because nonhuman primates (NHP) play a crucial role in modeling HIV (SIV) infections, it is crucial to be able to assess and characterize the NKG2 family in NHP. Unfortunately, it is not possible to detect CD94 using commercially available antibodies in NHP. Our work, a first for NHP, has focused on developing RNA flow cytometry using mRNA transcripts as proxies distinguishing NKG2C from NKG2A. We have expanded the application of this technology and here we show the first characterization of CD94 (KLRD1) NK cells in NHP using multiparametric RNA flow cytometry. Peripheral blood mononuclear cells from naive and matched acutely ( = 4) or chronically ( = 12) SIV-infected rhesus macaques were analyzed by flow cytometry using commercially available antibodies, determining expression of transcripts for NKG2A, NKG2C, and CD94 (KLRC1, KLRC2, and KLRD1, respectively) on NK cells using RNA flow cytometry. Our data show that KLRC1 KLRC2 KLRD1 NK cells decrease following chronic, but not acute, infection with SIV. This approach will allow us to investigate the kinetics of infection and NK memory formation and will further improve our understanding of basic NK cell biology, especially in the context of SIV infection. Nonhuman primates play a crucial role in approximating human biology and many diseases that are difficult, if not impossible, to achieve in other animal models, notably HIV. Current advances in adaptive NK cell research positions us to address fundamental deficiencies in our fight against infection and disease at the earliest moments after infection or substantially earlier in disease progression. We show here that we can identify specific NK cell subpopulations that are modulated following chronic, but not acute, SIV infection. The ability to identify these subsets more precisely will inform therapeutic and vaccine strategies targeting an optimized NK cell response.
最近，我们和其他人已经表明，自然杀伤 (NK) 细胞对巨细胞病毒 (CMV) 和人类免疫缺陷/病毒猴免疫缺陷病毒 (HIV/SIV) 感染表现出记忆样回忆反应。尽管其机制尚未完全阐明，但有几个小组已经表明，在恒河猴 CMV (rhCMV) 或人 CMV (hCMV) 感染的情况下，激活受体 NKG2C 在 NK 细胞上升高。与适应性 NK 细胞反应相关的 CD94 与 NKG2C 异二聚化。由于非人类灵长类动物 (NHP) 在模拟 HIV(SIV) 感染方面起着至关重要的作用，因此能够评估和表征 NHP 中的 NKG2 家族至关重要。不幸的是，不可能在 NHP 中使用市售抗体检测 CD94。我们的工作，这是 NHP 的首创，专注于使用 mRNA 转录本作为区分 NKG2C 与 NKG2A 的代表物开发 RNA 流式细胞术。我们已经扩展了这项技术的应用，并且在这里我们首次使用多参数 RNA 流式细胞术对 NHP 中的 CD94（KLRD1）NK 细胞进行了表征。使用商业上可用的抗体通过流式细胞术分析来自未感染和匹配的急性（= 4）或慢性（= 12）SIV 感染的恒河猴外周血单核细胞，使用 RNA 流式细胞术确定 NK 细胞上 NKG2A、NKG2C 和 CD94（KLRC1、KLRC2 和 KLRD1，分别）的转录物表达。我们的数据表明，KLRC1 KLRC2 KLRD1 NK 细胞在慢性而非急性 SIV 感染后减少。这种方法将使我们能够研究感染和 NK 记忆形成的动力学，并进一步提高我们对基本 NK 细胞生物学的理解，特别是在 SIV 感染的背景下。非人类灵长类动物在近似人类生物学方面起着至关重要的作用，并且在其他动物模型中难以甚至不可能实现许多疾病，特别是 HIV。适应性 NK 细胞研究的当前进展使我们能够在感染或疾病进展的早期阶段，在感染后最早的时刻解决感染和疾病的基本缺陷。我们在这里表明，我们可以识别在慢性而非急性 SIV 感染后发生调节的特定 NK 细胞亚群。更准确地识别这些亚群的能力将为针对优化的 NK 细胞反应的治疗和疫苗策略提供信息。
