Translational Sciences, Sanofi, Chilly-Mazarin, France.
Viral Pathogenesis and Biosafety Group, San Raffaele Scientific Institute, Milan, Italy.
Methods Mol Biol. 2022;2407:97-101. doi: 10.1007/978-1-0716-1871-4_9.
In addition to CD4 T cells, tissue-resident macrophages are target of productive HIV-1 infection. Unlike CD4 T lymphocytes they are characterized by a substantial resistance to the cytopathic effects triggered by viral infection. This feature, in addition to their homeostatic self-renewal capacity, strongly support the hypothesis that macrophages could serve as an additional reservoir of persistently infected cells in individuals receiving combination antiretroviral therapy (cART).In order to study the peculiar aspects of HIV-1 infection of macrophages, human primary monocyte-derived macrophages (MDM) represent the most exploited model given the difficulty to obtain and maintain in culture for significant periods of time macrophages from different organs and tissues. Here we present a model of MDM differentiation achieved in the absence of addition of exogenous cytokines (such as GM-CSF, discussed in the previous chapter), that could be further investigated in term of cell polarization toward classic, proinflammatory "M1", or alternatively activated "M2" cells before or after infection. We will also discuss how to reinforce the M1-polarization protocol to obtain a reliable model of reversible latency of infectious HIV-1 in primary M1-MDM.
除了 CD4 T 细胞，组织驻留巨噬细胞也是 HIV-1 感染的靶细胞。与 CD4 T 淋巴细胞不同，它们对病毒感染引发的细胞病变效应具有很强的抵抗力。除了它们的稳态自我更新能力外，这一特性强烈支持了这样一种假设，即巨噬细胞可以作为接受联合抗逆转录病毒治疗 (cART) 的个体中持续感染细胞的另一个储存库。为了研究 HIV-1 感染巨噬细胞的特殊方面，人原代单核细胞衍生的巨噬细胞 (MDM) 是最常用的模型，因为难以获得和维持不同器官和组织的巨噬细胞很长一段时间。在这里，我们提出了一种在不添加外源性细胞因子（如 GM-CSF，在前面的章节中讨论过）的情况下实现 MDM 分化的模型，该模型可以在感染前后进一步研究其向经典的、促炎的“M1”或替代性激活的“M2”细胞的极化。我们还将讨论如何加强 M1 极化方案，以获得在原代 M1-MDM 中可逆潜伏感染性 HIV-1 的可靠模型。
