VARIATIONS IN CD4 EXPRESSION BY HUMAN-MONOCYTES AND MACROPHAGES AND THEIR RELATIONSHIP TO INFECTION WITH THE HUMAN IMMUNODEFICIENCY VIRUS
VARIATIONS IN CD4 EXPRESSION BY HUMAN-MONOCYTES AND MACROPHAGES AND THEIR RELATIONSHIP TO INFECTION WITH THE HUMAN IMMUNODEFICIENCY VIRUS
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DOI：
10.1099/0022-1317-70-10-2661
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发表时间：
1989-10-01
影响因子：
3.8
通讯作者：
CUNNINGHAM, AL
中科院分区：
文献类型：
--
作者：
KAZAZI, F;MATHIJS, JM;CUNNINGHAM, AL
发现LeuM 3阳性人血液单核细胞表面上的CD 4抗原的表达是可变的，通过双染色流式细胞术，来自46名正常人志愿者的65 - 90%的细胞呈阳性。当单核细胞粘附在塑料上时（但在Teflon上培养时则不然），粘附后1至24 h观察到CD 4表达显著降低。用四种抗CD 4单克隆抗体在流式细胞术或直接免疫荧光法中不能检测到粘附于塑料5天的巨噬细胞中的CD 4表达。相反，在5天内表达LeuM 3和OKM 5表面抗原的贴壁细胞比例增加。CD 4 mRNA水平通过狭缝印迹和北方杂交测定，总细胞CD 4蛋白水平通过免疫沉淀测定。细胞mRNA和CD 4水平在整个5天期间保持恒定，但膜CD 4蛋白水平大大降低，表明CD 4的下调是翻译后的。两个六个新鲜的人类免疫缺陷病毒（HIV）分离株的感染表现出不同的动力学模式时，纯化的单核细胞最近粘附到塑料和巨噬细胞粘附5天。HIV抗原和逆转录酶水平在感染的单核细胞培养物中保持高水平3至4周，然后细胞发生脱落和坏死。巨噬细胞的感染产生低得多的抗原和逆转录酶水平，其在2周内下降到非常低或不可检测的水平，留下持续存活的巨噬细胞。感染后一周，在69 .+-. 7%单核细胞和6 . ±. 3%的巨噬细胞。用抗Leu 3a单克隆抗体进行的阻断实验表明，粘附5天的巨噬细胞的HIV感染主要通过与CD 4结合以外的机制发生。
The expression of CD4 antigen on the surface of LeuM3-positive human blood monocytes was found to be variable with 65 to 90% of cells from 46 normal human volunteers being positive by dual staining flow cytometry. When monocytes adhered to plastic (but not when cultured on Teflon), a marked decrease in CD4 expression was observed between 1 and 24 h post-adherence. CD4 expression could not be detected in macrophages adhered to plastic for 5 days by using four anti-CD4 monoclonal antibodies in flow cytometry or direct immunofluorescence. Conversely an increasing proportion of adherent cells expressed LeuM3 and OKM5 surface antigens over the 5 days. CD4 mRNA levels were measured by slot-blot and Northern hybridization, and total cellular CD4 protein levels by immunoprecipation. Both cellular mRNA and CD4 levels remained constant throughout the 5 day period but membrane CD4 protein levels were greatly reduced indicating that the down-regulation of CD4 was post-translational. Infection with two of six fresh human immunodeficiency virus (HIV) isolates showed different kinetic patterns when tested on purified monocytes recently adhered to plastic and macrophages adherent for 5 days. HIV antigen and reverse transcriptase levels in infected monocyte cultures remained high for 3 to 4 weeks before detachment and necrosis of the cells occurred. Infection of macrophages generated much lower levels of antigen and reverse transcriptase which declined to very low or undetectable levels over 2 weeks, leaving persisting viable macrophages. One week after infection HIV nucleic acid was detected in 69 .+-. 7% monocytes and 6 .+-. 3% of macrophages by in situ hybridization. Blocking experiments with anti-Leu3a monoclonal antibody suggested that HIV infection of 5 day adherent macrophages occurred mainly by a mechanism other than binding to CD4.