Quantitation of Productively Infected Monocytes and Macrophages of Simian Immunodeficiency Virus-Infected Macaques.
Quantitation of Productively Infected Monocytes and Macrophages of Simian Immunodeficiency Virus-Infected Macaques.
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DOI：
10.1128/jvi.00290-16
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发表时间：
2016-06-15
影响因子：
5.4
通讯作者：
Clements JE
中科院分区：
文献类型：
--
作者：
Avalos CR;Price SL;Forsyth ER;Pin JN;Shirk EN;Bullock BT;Queen SE;Li M;Gellerup D;O'Connor SL;Zink MC;Mankowski JL;Gama L;Clements JE
尽管联合抗逆转录病毒疗法（ART）取得了成功，但由于感染患者体内存在潜伏的病毒储库，人类免疫缺陷病毒（HIV）感染仍然是终身感染。CD 4 + T细胞对感染和疾病进展的贡献已被广泛研究。然而，在早期艾滋病毒感染期间，大脑和其他组织中的巨噬细胞会被感染，并导致组织特异性疾病，例如大脑中的脑炎和痴呆以及肺部的肺炎。单核细胞和巨噬细胞的感染程度尚未用与用于研究CD 4 + T细胞感染和评价携带感染性病毒基因组的CD 4 + T细胞数量的试验相当的试验进行严格评估。为了评估生产性感染的单核细胞和巨噬细胞对体内HIV和猴免疫缺陷病毒（SIV）感染细胞的贡献，我们基于用于定量HIV-和SIV-中的CD 4 + T细胞潜伏库的类似测定，开发了定量病毒生长测定（QVOA）。将表达CD 11b的髓样细胞连续稀释，并与易感的CD 11b细胞共培养。细胞来扩增病毒。测量T细胞受体β RNA作为对照，以评估CD 4 + T细胞在试验中的潜在贡献。通过定量逆转录-PCR定量上清液中的病毒产生。在血液、支气管肺泡灌洗液、肺、脾和脑中检测到有效感染的髓样细胞，表明这些细胞在SIV感染期间持续存在，并且在ART期间具有促成病毒储库的潜力。重要性CD 4 + T细胞的感染及其作为潜伏储库的作用已被严格评估;然而，还没有类似地研究体内产生性感染的单核细胞和巨噬细胞的频率。与淋巴细胞不同，髓样细胞对HIV的致细胞病变作用有抵抗力。此外，组织驻留巨噬细胞具有自我更新的能力，并在体内持续数月至数年。因此，组织巨噬细胞一旦被感染，就具有潜在稳定的病毒储存库的特征。更好地了解生产性感染的巨噬细胞的数量是至关重要的，以进一步评估感染的骨髓细胞作为一个潜在的病毒库的作用。在本研究中，我们比较了SIV感染猕猴模型中CD 4 + T细胞和巨噬细胞的感染频率。我们开发了一种关键的检测方法，使我们能够定量含有病毒基因组的骨髓细胞，这些病毒基因组导致SIV感染的猕猴产生感染，并评估巨噬细胞作为潜在储库的作用。
Despite the success of combined antiretroviral therapy (ART), human immunodeficiency virus (HIV) infection remains a lifelong infection because of latent viral reservoirs in infected patients. The contribution of CD4+ T cells to infection and disease progression has been extensively studied. However, during early HIV infection, macrophages in brain and other tissues are infected and contribute to tissue-specific diseases, such as encephalitis and dementia in brain and pneumonia in lung. The extent of infection of monocytes and macrophages has not been rigorously assessed with assays comparable to those used to study infection of CD4+ T cells and to evaluate the number of CD4+ T cells that harbor infectious viral genomes. To assess the contribution of productively infected monocytes and macrophages to HIV- and simian immunodeficiency virus (SIV)-infected cells in vivo, we developed a quantitative virus outgrowth assay (QVOA) based on similar assays used to quantitate CD4+ T cell latent reservoirs in HIV- and SIV-infected individuals in whom the infection is suppressed by ART. Myeloid cells expressing CD11b were serially diluted and cocultured with susceptible cells to amplify virus. T cell receptor β RNA was measured as a control to assess the potential contribution of CD4+ T cells in the assay. Virus production in the supernatant was quantitated by quantitative reverse transcription-PCR. Productively infected myeloid cells were detected in blood, bronchoalveolar lavage fluid, lungs, spleen, and brain, demonstrating that these cells persist throughout SIV infection and have the potential to contribute to the viral reservoir during ART. IMPORTANCE Infection of CD4+ T cells and their role as latent reservoirs have been rigorously assessed; however, the frequency of productively infected monocytes and macrophages in vivo has not been similarly studied. Myeloid cells, unlike lymphocytes, are resistant to the cytopathic effects of HIV. 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 potentially stable viral reservoir. A better understanding of the number of productively infected macrophages is crucial to further evaluate the role of infected myeloid cells as a potential viral reservoir. In the study described here we compared the frequency of productively infected CD4+ T cells and macrophages in an SIV-infected macaque model. We developed a critical assay that will allow us to quantitate myeloid cells containing viral genomes that lead to productive infection in SIV-infected macaques and assess the role of macrophages as potential reservoirs.