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关于口腔上皮细胞和朗格汉斯细胞作为潜在HIV病毒储存库作用的研究。

An investigation of the role of oral epithelial cells and Langerhans cells as possible HIV viral reservoirs.

作者信息

Boy Sonja C, van Heerden Marlene B, Wolfaardt Marianne, Cockeran Riana, Gema Elize, van Heerden Willie F

机构信息

Department of Oral Pathology and Oral Biology, School of Dentistry, University of Pretoria, Pretoria, South Africa.

出版信息

J Oral Pathol Med. 2009 Jan;38(1):114-9. doi: 10.1111/j.1600-0714.2008.00711.x.

Abstract

BACKGROUND

The role of the oral mucosa as a target of human immunodeficiency virus (HIV-1) infection and persistence is unclear. HIV-1 has been reported in oral epithelial cells, but this has not been confirmed. Cellular reservoirs may impede antiretroviral therapies and should be identified. This study was performed to determine the presence of HIV-1 in oral epithelial and Langerhans cells (LCs) of HIV-1-positive antiretroviral naïve patients. Non-invasive brush biopsy technique for future in vivo HIV research was also evaluated.

METHODS

Oral mucosal cells were harvested from the buccal mucosae, dorsal tongue and the gingiva of the mandibular teeth of 35 HIV-1-positive patients using a Cytobrush Plus cell collector. Epithelial cells were purified from the samples by flow cytometric cell sorting using cytokeratin stains after which the epithelial cell samples were further purified and divided into superficial and deep epithelial cells by laser microdissection on Pap stained cytospin smears. LCs were picked up individually by laser microdissection from CD1a stained cytospin smears. Purified epithelial and LC samples were tested for the presence of HIV-1 DNA by polymerase chain reaction analysis.

RESULTS

Ten of the patients had HIV-1 DNA in one or more of the sampled anatomical locations. No HIV-1 DNA could be demonstrated in any of the purified superficial or deep epithelial or LC samples.

CONCLUSIONS

HIV-DNA can be found using non-invasive oral brush biopsies and should be investigated further as an experimental model for in vivo oral HIV research. Better ways to purify the different cell types should be investigated.

摘要

背景

口腔黏膜作为人类免疫缺陷病毒(HIV-1)感染和持续存在的靶点,其作用尚不清楚。已有报道称在口腔上皮细胞中发现了HIV-1,但尚未得到证实。细胞储存库可能会阻碍抗逆转录病毒疗法,因此需要加以识别。本研究旨在确定HIV-1阳性且未接受过抗逆转录病毒治疗的患者口腔上皮细胞和朗格汉斯细胞(LCs)中是否存在HIV-1。还评估了用于未来体内HIV研究的非侵入性刷检活检技术。

方法

使用Cytobrush Plus细胞采集器从35例HIV-1阳性患者的颊黏膜、舌背和下颌牙龈采集口腔黏膜细胞。使用细胞角蛋白染色通过流式细胞术细胞分选从样本中纯化上皮细胞,之后在巴氏染色的细胞涂片上通过激光显微切割将上皮细胞样本进一步纯化并分为浅表和深层上皮细胞。通过激光显微切割从CD1a染色的细胞涂片上逐个挑选LCs。通过聚合酶链反应分析检测纯化后的上皮细胞和LCs样本中是否存在HIV-1 DNA。

结果

10例患者在一个或多个采样解剖部位检测到HIV-1 DNA。在任何纯化后的浅表或深层上皮细胞或LCs样本中均未检测到HIV-1 DNA。

结论

使用非侵入性口腔刷检活检可发现HIV-DNA,作为体内口腔HIV研究的实验模型应进一步研究。应探索更好的方法来纯化不同的细胞类型。

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