Efficient isolation and propagation of human immunodeficiency virus on recombinant colony-stimulating factor 1-treated monocytes.
Efficient isolation and propagation of human immunodeficiency virus on recombinant colony-stimulating factor 1-treated monocytes.
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人类免疫缺陷病毒对重组刺激因子1处理的单核细胞的有效分离和传播。
DOI：
10.1084/jem.167.4.1428
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发表时间：
1988-04-01
影响因子：
15.3
通讯作者：
MELTZER, MS
中科院分区：
文献类型：
--
作者：
GENDELMAN, HE;ORENSTEIN, JM;MARTIN, MA;FERRUA, C;MITRA, R;PHIPPS, T;WAHL, LA;LANE, HC;FAUCI, AS;BURKE, DS;SKILLMAN, D;MELTZER, MS
单核细胞在补充有rCSF-1的培养基中在组织培养物中维持超过3个月。这些培养物为从HIV感染患者的PBMC中分离和增殖病毒提供了易感靶细胞。分离到单核细胞中的HIV容易感染其他rCSF-1处理的单核细胞，但只能无效地感染PHA刺激的淋巴母细胞。类似地，在T细胞系中传代的实验室HIV毒株或从患者的白细胞中分离的病毒进入PHA刺激的成淋巴细胞中，不能有效地感染rCSF-1处理的单核细胞。通过6-7周的逆转录酶活性或HIV抗原捕获，在巨噬细胞培养液中检测到持续的低水平病毒体产生。感染后2周，单核细胞培养物中观察到细胞形态的显著变化，包括细胞死亡、合胞体和巨细胞形成，但在4-6周时，所有细胞形态均正常。然而，在这些培养物中，在6周时感染细胞的频率为60-90%，如通过与HIV RNA探针的原位杂交或通过与AIDS患者血清的免疫荧光所定量的。电镜超微结构分析也显示了高频率的感染细胞，几乎所有的艾滋病毒出芽到细胞质空泡和病毒颗粒内积累，只有很少与质膜。病毒在巨噬细胞内的滞留和HIV变异体的巨噬细胞嗜性可以解释疾病期间病毒持续存在和传播的机制。
Monocytes were maintained in tissue culture for greater than 3 mo in media supplemented with rCSF-1. These cultures provided susceptible target cells for isolation and propagation of virus from PBMC of HIV- infected patients. HIV isolated into monocytes readily infected other rCSF-1-treated monocytes but only inefficiently infected PHA-stimulated lymphoblasts. Similarly, laboratory HIV strains passaged in T cell lines or virus isolated from patients' leukocytes into PHA-stimulated lymphoblasts inefficiently infected rCSF-1-treated monocytes. Persistent, low-level virion production was detected in macrophage culture fluids by reverse transcriptase activity or HIV antigen capture through 6-7 wk. Marked changes in cell morphology with cell death, syncytia, and giant cell formation were observed in monocyte cultures 2 wk after infection, but at 4-6 wk, all cells appeared morphologically normal. However, the frequency of infected cells in these cultures at 6 wk was 60-90% as quantified by in situ hybridization with HIV RNA probes or by immunofluorescence with AIDS patients' sera. Ultrastructural analysis by EM also showed a high frequency of infected cells; virtually all HIV budded into and accumulated within cytoplasmic vacuoles and virus particles were only infrequently associated with the plasma membrane. Retention of virus within macrophages and the macrophage tropism of HIV variants may explain mechanisms of both virus persistence and dissemination during disease.
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影响因子：
56.9
作者：
BARRESINOUSSI, F;CHERMANN, JC;MONTAGNIER, L
通讯作者：
MONTAGNIER, L
影响因子：
56.9
作者：
CLARK, SC;KAMEN, R
通讯作者：
KAMEN, R
DOI：
10.1073/pnas.82.20.7086
发表时间：
1985-01-01
影响因子：
11.1
作者：
GENDELMAN, HE;NARAYAN, O;GHOTBI, Z
通讯作者：
GHOTBI, Z
影响因子：
56.9
作者：
CLAVEL, F;GUETARD, D;MONTAGNIER, L
通讯作者：
MONTAGNIER, L
影响因子：
56.9
作者：
KOENIG, S;GENDELMAN, HE;FAUCI, AS
通讯作者：
FAUCI, AS