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人表皮朗格汉斯细胞的HIV-1体外感染

In vitro infection of human epidermal Langerhans' cells with HIV-1.

作者信息

Ramazzotti E, Marconi A, Re M C, Girolomoni G, Cenacchi G, Vignoli M, Zambruno G, Furlini G, La Placa M, Giannetti A

机构信息

Institute of Microbiology, University of Bologna, Italy.

出版信息

Immunology. 1995 May;85(1):94-8.

Abstract

Epidermal Langerhans' cells (LC) from human immunodeficiency virus type-1 (HIV-1)-infected patients harbour HIV-1 proviral DNA and RNA. In the present study, we investigated whether LC from epidermis of normal, HIV-seronegative subjects could be infected in vitro with HIV-1. Epidermal cells (EC) spontaneously detached from epidermal sheet cultures were enriched for LC (10-25% of CD1a+/CD4+ cells), deprived of contaminating T cells and then incubated with HIV-1IIIB. After 24 hr, purified LC and LC-depleted EC fractions were obtained by immunomagnetic separation. Polymerase chain reaction (PCR) analysis showed the presence of HIV-1 proviral DNA (gag) only in purified LC. In addition, LC-enriched EC, purified LC, LC-depleted EC or the non-permissive cell line, TF-1, the latter having being previously challenged with HIV-1IIIB for the same length of time as the EC, were co-cultivated with C8166 cells, and the co-cultures assessed for the presence of HIV DNA by PCR. Co-cultures of C8166 cells with purified LC or LC-enriched EC previously exposed to HIV-1IIIB exhibited a time-dependent increase in HIV proviral DNA. In contrast, PCR analysis of C8166 cells co-cultured with either LC-depleted EC or TF-1 cells gave negative results. Finally, C8166 cells co-cultured with HIV-infected LC formed syncytia, showed membrane budding and released numerous retroviral particles. The results indicate that LC from normal subjects can be infected in vitro with HIV and can transmit infection to myeloid cells. This in vitro model may help in understanding the regulation of HIV infection of LC.

摘要

来自人类免疫缺陷病毒1型(HIV-1)感染患者的表皮朗格汉斯细胞(LC)携带HIV-1前病毒DNA和RNA。在本研究中,我们调查了来自正常、HIV血清学阴性受试者表皮的LC在体外是否能被HIV-1感染。从表皮片培养物中自发脱离的表皮细胞(EC)富集了LC(占CD1a+/CD4+细胞的10 - 25%),去除污染的T细胞后,再与HIV-1IIIB一起孵育。24小时后,通过免疫磁珠分离获得纯化的LC和去除LC的EC组分。聚合酶链反应(PCR)分析显示,仅在纯化的LC中存在HIV-1前病毒DNA(gag)。此外,将富集LC的EC、纯化的LC、去除LC的EC或非允许细胞系TF-1(后者先前已用HIV-1IIIB攻击相同时间,与EC相同)与C8166细胞共培养,并通过PCR评估共培养物中HIV DNA的存在情况。C8166细胞与先前暴露于HIV-1IIIB的纯化LC或富集LC的EC的共培养物中,HIV前病毒DNA呈时间依赖性增加。相反,与去除LC的EC或TF-1细胞共培养的C8166细胞的PCR分析结果为阴性。最后,与HIV感染的LC共培养的C8166细胞形成了多核巨细胞,出现膜出芽并释放了大量逆转录病毒颗粒。结果表明,正常受试者的LC在体外可被HIV感染,并能将感染传播给髓系细胞。这个体外模型可能有助于理解LC的HIV感染调控机制。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/0f6c/1384030/ad9da3321d06/immunology00067-0103-a.jpg

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