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体外离心增强人原代单核细胞/巨噬细胞中1型人类免疫缺陷病毒感染及人巨细胞病毒基因表达

Centrifugal enhancement of human immunodeficiency virus type 1 infection and human cytomegalovirus gene expression in human primary monocyte/macrophages in vitro.

作者信息

Ho W Z, Cherukuri R, Ge S D, Cutilli J R, Song L, Whitko S, Douglas S D

机构信息

Division of Allergy, Immunology, and Infectious Diseases, Children's Hospital of Philadelphia, Pennsylvania 19104.

出版信息

J Leukoc Biol. 1993 Feb;53(2):208-12. doi: 10.1002/jlb.53.2.208.

Abstract

In an effort to facilitate the efficiency of human immunodeficiency virus type 1 (HIV-1) and/or human cytomegalovirus (HCMV) infection in primary monocyte/macrophages in vitro, the effect of low-speed centrifugation was studied. The infectivity of three strains (Bal, Ada-M, and IIIB) of HIV-1 tested was significantly enhanced by centrifugal inoculation at a force of 1500g for 60 min. Reverse transcriptase activity and HIV-1 p24 antigen in primary monocyte/macrophages infected by a centrifugal inoculation technique were detectable 3-7 days earlier and were more than 10-fold greater in magnitude (at an early stage of the infection) than those of control cells infected by the conventional inoculation technique. Examination of the cells by indirect immunofluorescence revealed higher expression of HIV-1 p24 protein in the monocyte/macrophages infected by the centrifugal inoculation technique. These differences were directly related to centrifugal inoculation and were evident up to 3 weeks after infection. Enhancement was not observed when centrifugation was carried out before or after HIV-1 infection. Centrifugal inoculation of HCMV also enhanced its immediate-early and early gene expression up to 30- to 50-fold, although neither late nuclear antigens and glycoproteins of HCMV nor infectious virus was detected in HCMV-infected monocyte/macrophage cultures. These results show that centrifugal inoculation is a useful technique for improving the efficiency of HCMV and HIV-1 infection in vitro.

摘要

为了提高人免疫缺陷病毒1型(HIV-1)和/或人巨细胞病毒(HCMV)在原代单核细胞/巨噬细胞中的体外感染效率,研究了低速离心的作用。通过以1500g的力离心接种60分钟,所测试的三种HIV-1毒株(Bal、Ada-M和IIIB)的感染性显著增强。采用离心接种技术感染的原代单核细胞/巨噬细胞中的逆转录酶活性和HIV-1 p24抗原比采用传统接种技术感染的对照细胞可提前三至七天检测到,且(在感染早期)其水平高出十倍以上。通过间接免疫荧光检查细胞发现,采用离心接种技术感染的单核细胞/巨噬细胞中HIV-1 p24蛋白的表达更高。这些差异与离心接种直接相关,且在感染后三周内都很明显。在HIV-1感染之前或之后进行离心均未观察到增强作用。HCMV的离心接种也使其立即早期和早期基因表达增强了30至50倍,尽管在感染HCMV的单核细胞/巨噬细胞培养物中未检测到HCMV的晚期核抗原、糖蛋白和感染性病毒。这些结果表明,离心接种是一种提高HCMV和HIV-1体外感染效率的有用技术。

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