Besansky N J, Butera S T, Sinha S, Folks T M
Retrovirus Diseases Branch, Centers for Disease Control, Atlanta, Georgia 30333.
J Virol. 1991 May;65(5):2695-8. doi: 10.1128/JVI.65.5.2695-2698.1991.
Using a polymerase chain reaction-based assay on total cell lysates, we have detected unintegrated human immunodeficiency virus type 1 (HIV-1) DNA in chronically infected T-lymphocytic (ACH-2, J1) and promyelocytic (OM-10.1) cell lines. Treatment with 3'-azido-3'-deoxythymidine (AZT) or soluble CD4 inhibited accumulation of unintegrated viral DNA about 10-fold within 72 h; removal of AZT permitted recovery to pretreatment levels within 72 h. Our results indicate that unintegrated HIV-1 DNA is unstable in these cell lines and originates from a continuous process of reinfection. OM-10.1 cells had relatively high levels of surface CD4 by flow cytometry and high levels of unintegrated viral DNA by polymerase chain reaction. ACH-2 cells had very low levels of both surface CD4 and unintegrated viral DNA. However, J1 cells, with surface CD4 below the level of detection of flow cytometry had a high level of unintegrated viral DNA similar to that of OM-10.1 cells. This implies that the number of CD4 receptors is not rate limiting for reinfection.
通过对全细胞裂解物进行基于聚合酶链反应的检测,我们在慢性感染的T淋巴细胞系(ACH-2、J1)和早幼粒细胞系(OM-10.1)中检测到了未整合的1型人类免疫缺陷病毒(HIV-1)DNA。用3'-叠氮-3'-脱氧胸苷(AZT)或可溶性CD4处理可在72小时内将未整合病毒DNA的积累抑制约10倍;去除AZT可使病毒DNA在72小时内恢复到预处理水平。我们的结果表明,未整合的HIV-1 DNA在这些细胞系中不稳定,且源自持续的再感染过程。通过流式细胞术检测,OM-10.1细胞表面CD4水平相对较高,通过聚合酶链反应检测其未整合病毒DNA水平也较高。ACH-2细胞表面CD4和未整合病毒DNA水平都非常低。然而,J1细胞表面CD4低于流式细胞术的检测水平,但其未整合病毒DNA水平却与OM-10.1细胞相似,很高。这意味着CD4受体的数量不是再感染的限速因素。