Weeks B S, Klotman M E, Dhawan S, Kibbey M, Rappaport J, Kleinman H K, Yamada K M, Klotman P E
Laboratory of Developmental Biology, National Institute of Dental Research, National Institutes of Health, Bethesda, Maryland 20892.
J Cell Biol. 1991 Aug;114(4):847-53. doi: 10.1083/jcb.114.4.847.
Altered T cell adherence after human immunodeficiency virus 1 (HIV-1) infection may contribute to viral pathogenesis in the acquired immune deficiency syndrome. To address this hypothesis, we assessed mechanisms of T cell adherence to extracellular matrix proteins in vitro. We found that after HIV-1 infection, both chronically infected H9 CD4+ T cells and acutely infected primary peripheral blood lymphocytes acquired the ability to adhere to the extracellular matrix glycoprotein fibronectin, to a lesser extent to type IV collagen and laminin, but not to type I collagen. H9 cells chronically infected with two of the three HIV-1 strains studied showed approximately a sevenfold increase in attachment to fibronectin, while the same cells infected with the human retrovirus HIV-2 did not. Adhesion was accompanied by changes in morphology, including marked spreading and increased filopodia. These alterations were not blocked by the protein kinase C inhibitor H-7, which did inhibit TPA-induced T cell attachment to fibronectin. Monoclonal antibodies against both the alpha 5 and the beta 1 subunits of the classical fibronectin receptor as well as an Arg-Gly-Asp (RGD) peptide inhibited attachment, whereas anti-alpha 4 monoclonal antibodies and the CS1 peptide did not. Binding to collagen IV was also inhibited by the anti-beta 1 monoclonal antibody, but not the other antibodies. Cells metabolically labeled with [35S]methionine and analyzed by immunoprecipitation with polyclonal anti-beta 1 integrin antibody showed a 2.5-fold increase in integrin synthesis in infected cells compared to uninfected controls. This increase in synthesis was associated with an increase in cell surface expression of both alpha 5 and beta 1 integrins by FACS (registered trademark of Becton Dickinson for a fluorescence-activated cell sorter) analysis. Enhanced expression of integrins such as alpha 5 beta 1 may cause T cell adherence to a variety of tissues, where released viral gene products may induce some of the tissue-specific manifestations of HIV-1 infection.
人类免疫缺陷病毒1型（HIV-1）感染后T细胞黏附的改变可能在获得性免疫缺陷综合征的病毒发病机制中起作用。为了验证这一假说，我们在体外评估了T细胞黏附细胞外基质蛋白的机制。我们发现，HIV-1感染后，慢性感染的H9 CD4+ T细胞和急性感染的原代外周血淋巴细胞均获得了黏附细胞外基质糖蛋白纤连蛋白的能力，对IV型胶原和层粘连蛋白的黏附能力较弱，但对I型胶原无黏附能力。用三种研究的HIV-1毒株中的两种慢性感染的H9细胞对纤连蛋白的黏附增加了约7倍，而感染人类逆转录病毒HIV-2的相同细胞则没有。黏附伴随着形态学变化，包括明显的铺展和丝状伪足增加。这些改变未被蛋白激酶C抑制剂H-7阻断，而H-7确实抑制了佛波酯诱导的T细胞对纤连蛋白的黏附。针对经典纤连蛋白受体的α5和β1亚基的单克隆抗体以及精氨酸-甘氨酸-天冬氨酸（RGD）肽均抑制黏附，而抗α4单克隆抗体和CS1肽则无此作用。抗β1单克隆抗体也抑制了与IV型胶原的结合，但其他抗体则无此作用。用[35S]甲硫氨酸进行代谢标记并用多克隆抗β1整合素抗体进行免疫沉淀分析的细胞显示，与未感染对照相比，感染细胞中的整合素合成增加了2.5倍。这种合成增加与通过荧光激活细胞分选仪（FACS，Becton Dickinson公司的注册商标）分析的α5和β1整合素在细胞表面表达的增加有关。诸如α5β1等整合素的表达增强可能导致T细胞黏附于多种组织，在这些组织中释放的病毒基因产物可能引发HIV-1感染的一些组织特异性表现。
