Autissier Estelle, Li Haiying, Goepfert Paul A, Reeves R Keith
1 Division of Immunology, New England Primate Research Center , Harvard Medical School, Southborough, Massachusetts.
2 Center for Virology and Vaccine Research, Beth Israel Deaconess Medical Center , Harvard Medical School, Boston, Massachusetts.
AIDS Res Hum Retroviruses. 2018 May;34(5):446-448. doi: 10.1089/AID.2018.0011. Epub 2018 Mar 27.
Apoptotic membrane microparticles (MMPs) derived from dying cells of multiple cell origins are highly immunostimulatory and are indicative of global immune activation and cell death in a variety of diseases. In this study, we developed a flow cytometric bead assay to quantify annexin-V apoptotic (MMPs) in plasma from humans and rhesus macaques. With a combination of flow cytometry and pan-fluorescent beads, MMPs were enumerated in plasma specimens by adding a constant ratio of beads to initial fluid volumes and then calculating MMP/mL based on MMP-to-bead ratios. Using this straightforward assay, we found that circulating MMP quantifications were highly reproducible and similar in number between normal rhesus macaques and humans subjects. However, MMPs increased two- to threefold during HIV and simian immunodeficiency virus (SIV) infections and were positively associated with T cell immune activation. Collectively, we present a rapid bead-based assay for both humans and macaque models to quantify MMPs that could be an instigator and predictor of immune activation, which is a primary source of HIV/SIV disease.
源自多种细胞来源的濒死细胞的凋亡膜微粒(MMPs)具有高度免疫刺激作用,并且在多种疾病中指示全身免疫激活和细胞死亡。在本研究中,我们开发了一种流式细胞术磁珠检测法,用于定量人和恒河猴血浆中的膜联蛋白-V凋亡(MMPs)。通过将流式细胞术与全荧光磁珠相结合,在血浆样本中通过向初始液体体积中加入恒定比例的磁珠来计数MMPs,然后根据MMP与磁珠的比例计算每毫升中的MMP数量。使用这种简单的检测方法,我们发现正常恒河猴和人类受试者之间循环MMP的定量具有高度可重复性且数量相似。然而,在HIV和猴免疫缺陷病毒(SIV)感染期间,MMPs增加了两到三倍,并且与T细胞免疫激活呈正相关。总体而言,我们提出了一种针对人类和猕猴模型的基于磁珠的快速检测法,用于定量MMPs,MMPs可能是免疫激活的引发剂和预测指标,而免疫激活是HIV/SIV疾病的主要来源。