St Luce S, Arts E, Geleziunas R, Bour S, Wainberg M A
Lady Davis Institute, Jewish General Hospital, Montreal, Quebec, Canada.
J Med Virol. 1993 Sep;41(1):71-8. doi: 10.1002/jmv.1890410115.
We have infected ten-day-old primary cultures of human monocyte-derived macrophages (MDM) with HIV-1 by cocultivation with chronically infected monocytic cell lines. This work has involved the U-937 monocytoid cell line, chronically infected with the HIV-IIIB strain of HIV-1 (U-937HIV IIIB) as well as a number of cell clones, termed UHC, which were derived from U-937HIV IIIB by limiting dilution. Cell-free virus, derived from each of U-937HIV IIIB cells and the UHC1 clone were noninfectious for MDM, as determined by failure to express viral p24 antigen (Ag). In contrast, viral p24 Ag production was detected in MDM that had been cocultivated with U-937HIV IIB, and with each of three UHC clones that produced infectious virus. Infection, in each case, was confirmed by polymerase chain reaction detection via the amplification of proviral DNA. In contrast, cocultivation with the UHC15.7 clone, which fails to cleave viral gp160 to its gp120 and gp41 products or the UHC8 clone, which lacks functional reverse transcriptase, did not lead to infection of MDM. Pretreatment of MDM for 2 hr with 1 microM AZT completely prevented infection by culture fluids containing HIVada, a macrophage-tropic virus, but did not affect infection mediated by cocultivation. These results suggest that cell-to-cell transmission of HIV-1, among monocyte-derived macrophages, can be mediated by proviral DNA. Moreover, gp120 at the surface of infected cells may play an important role in this process, since cell-to-cell HIV transmission could not be demonstrated with the UHC clone that is defective in cleavage of the viral envelope glycoprotein gp160.
我们通过与慢性感染的单核细胞系共培养，用HIV-1感染了人单核细胞衍生巨噬细胞（MDM）的10日龄原代培养物。这项工作涉及长期感染HIV-1的HIV-IIIB株的U-937单核细胞样细胞系（U-937HIV IIIB）以及一些细胞克隆，称为UHC，它们是通过有限稀释从U-937HIV IIIB衍生而来的。从U-937HIV IIIB细胞和UHC1克隆中获得的无细胞病毒对MDM无感染性，这是通过未能表达病毒p24抗原（Ag）来确定的。相比之下，在与U-937HIV IIB以及产生感染性病毒的三个UHC克隆中的每一个共培养的MDM中检测到了病毒p24 Ag的产生。在每种情况下，感染都通过聚合酶链反应检测前病毒DNA的扩增得到证实。相比之下，与未能将病毒gp160切割成其gp120和gp41产物的UHC15.7克隆或缺乏功能性逆转录酶的UHC8克隆共培养，并未导致MDM感染。用1 microM叠氮胸苷（AZT）对MDM预处理2小时可完全阻止由含有巨噬细胞嗜性病毒HIVada的培养液引起的感染，但不影响共培养介导的感染。这些结果表明，HIV-1在单核细胞衍生巨噬细胞之间的细胞间传播可由前病毒DNA介导。此外，感染细胞表面的gp120可能在这一过程中起重要作用，因为用在病毒包膜糖蛋白gp160切割方面有缺陷的UHC克隆无法证明HIV的细胞间传播。
