Creson J R, Lin A A, Li Q, Broad D F, Roberts M R, Anderson S J
Cell Genesys, Inc., Foster City, California 94404, USA.
J Virol. 1999 Nov;73(11):9337-47. doi: 10.1128/JVI.73.11.9337-9347.1999.
We have investigated the ability of anti-CD28 antibody costimulation to induce resistance to macrophage (M)-tropic strains of human immunodeficiency virus type 1 (HIV-1) in vitro. Our results confirm the observations of Levine et al. (15) that stimulation of CD4 T cells with anti-CD3/anti-CD28 antibodies coimmobilized on magnetic beads renders the cells resistant to infection by M-tropic strains of HIV-1. The resistance was strongest when the beads were left in the cultures throughout the experiment. In contrast, stimulation of CD4 T cells with the same antibodies immobilized on the surface of plastic culture dishes failed to induce resistance and resulted in high levels of p24 production. This was true even if the cells were passaged continuously on freshly coated plates. If the beads were removed after initial stimulation, p24 production increased over time and produced a result intermediate to the other forms of stimulation. For beads-in, beads-out, and one-time plate stimulated cultures, resistance to infection correlated with down-regulation of CCR5 expression at the cell surface and with increased production of beta-chemokines. However, cultures of CD4 T cells continuously passaged on anti-CD3/anti-CD28-coated plates produced large amounts of p24 despite decreased levels of CCR5 expression and increasing production of beta-chemokines. Expression of the T-cell activation markers CD25 and CD69 and production of gamma interferon further supported the differences in plate versus bead stimulation. Our results explain the apparent contradiction between the ability of anti-CD28 antibody costimulation to induce resistance to HIV infection when presented on magnetic beads and the increased ability to recover virus from the cells of HIV-positive donors who are on highly active antiretroviral therapy when cells are stimulated by anti-CD3/anti-CD28 immobilized on plastic dishes.
我们研究了抗CD28抗体共刺激在体外诱导对1型人类免疫缺陷病毒（HIV-1）巨噬细胞（M）嗜性毒株产生抗性的能力。我们的结果证实了Levine等人（15）的观察结果，即固定在磁珠上的抗CD3/抗CD28抗体刺激CD4 T细胞可使细胞对HIV-1的M嗜性毒株感染产生抗性。当磁珠在整个实验过程中都留在培养物中时，抗性最强。相比之下，用固定在塑料培养皿表面的相同抗体刺激CD4 T细胞未能诱导抗性，并导致高水平的p24产生。即使细胞在新包被的培养皿上连续传代，情况也是如此。如果在初始刺激后去除磁珠，p24产量会随时间增加，并产生介于其他刺激形式之间的结果。对于磁珠存在、磁珠去除和一次性平板刺激的培养物，对感染的抗性与细胞表面CCR5表达的下调以及β趋化因子产生的增加相关。然而，在抗CD3/抗CD28包被的平板上连续传代的CD4 T细胞培养物尽管CCR5表达水平降低且β趋化因子产生增加，但仍产生大量p24。T细胞活化标志物CD25和CD69的表达以及γ干扰素的产生进一步支持了平板刺激与磁珠刺激之间的差异。我们的结果解释了抗CD28抗体共刺激在磁珠上呈现时诱导对HIV感染产生抗性的能力与在接受高效抗逆转录病毒治疗的HIV阳性供体的细胞中，当细胞由固定在塑料培养皿上的抗CD3/抗CD28刺激时从细胞中回收病毒的能力增加之间明显的矛盾。
