Department of Molecular and Comparative Pathobiology, Johns Hopkins University School of Medicine, Baltimore, MD, USA.
Methods Mol Biol. 2022;2407:333-356. doi: 10.1007/978-1-0716-1871-4_22.
The role of CD4+ T cells in HIV infection and the latent reservoir, that is, latently infected cells that harbor replication competent virus, has been rigorously assessed. We have previously reported a quantitative viral outgrowth assay (QVOA) for SIV that demonstrated the frequency of latently infected CD4+ T cells is approximately 1 in a million cells, similar to that of HIV infected individuals on ART. However, the frequency of productively infected monocytes in blood and macrophages in tissues has not been similarly studied. Myeloid cells are infected during acute HIV and SIV infection; however, unlike lymphocytes, they are resistant to the cytopathic effects of the virus. Moreover, tissue-resident macrophages have the ability to self-renew and persist in the body for months to years. Thus, tissue macrophages, once infected, have the characteristics of a stable viral reservoir. A better understanding of the number of productively infected macrophages is critical to understanding the role of infected myeloid cells as a viral reservoir. In order to assess the functional latent reservoir. we have developed specific QVOAs for monocytes in blood, and macrophages in spleen, BAL and brain, which are described in detail in this chapter.
CD4+T 细胞在 HIV 感染和潜伏储库（即潜伏感染的细胞含有具有复制能力的病毒）中的作用已经得到了严格评估。我们之前报道了一种用于 SIV 的定量病毒扩增检测（QVOA），该检测表明潜伏感染的 CD4+T 细胞的频率约为每百万细胞中有一个，类似于接受 ART 的 HIV 感染者。然而，关于血液中受感染的单核细胞和组织中的巨噬细胞的感染频率尚未进行类似研究。髓样细胞在急性 HIV 和 SIV 感染期间被感染；然而，与淋巴细胞不同，它们对病毒的细胞病变效应具有抗性。此外，组织驻留巨噬细胞具有自我更新和在体内持续数月至数年的能力。因此，一旦感染，组织巨噬细胞就具有稳定的病毒储库的特征。更好地了解受感染的巨噬细胞数量对于理解感染的髓样细胞作为病毒储库的作用至关重要。为了评估功能性潜伏储库，我们已经针对血液中的单核细胞和脾脏、BAL 和大脑中的巨噬细胞开发了特定的 QVOA，本章详细描述了这些方法。
