Vaginal transmission of HIV-1 in hu-SCID mice: a new model for the evaluation of vaginal microbicides
Vaginal transmission of HIV-1 in hu-SCID mice: a new model for the evaluation of vaginal microbicides
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DOI：
10.1097/00002030-200111230-00003
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发表时间：
2001-11-23
期刊：
影响因子：
3.8
通讯作者：
Vella, S
中科院分区：
文献类型：
--
作者：
Di Fabio, S;Giannini, G;Vella, S
目的：建立HIV-1阴道传播动物模型，评价阴道杀菌剂的作用。设计：通过无创阴道给药，用4 × 10(7)人外周血淋巴细胞（hu-PBL）重组SCID小鼠进行阴道感染。hu-PBL先前在体外用HIV-1 (SF162) （hu-PBL- scid）的非合胞体（NSI）菌株感染。用荧光标记的人淋巴细胞检测体内淋巴细胞的迁移。方法：分别采用荧光活化细胞分选（FACS）、Amplicor HIV-1监测试剂盒和酶联免疫吸附法检测CD4 T细胞百分比、血浆病毒载量和p24抗原。对脾脏和淋巴结提取的DNA进行聚合酶链反应（PCR）分析。标记淋巴细胞的体内迁移，分别于4、24、48 h后处死小鼠；切除阴道和局部淋巴结，OCT快速冷冻，荧光显微镜和FACS切片检查。结果：通过FACS、HIV病毒载量、p24和PCR检测，证实经阴道接种的病毒感染细胞存在HIV传播。标记的细胞在4 h后很容易定位到阴道组织内，但在24或48 h后阴道水平几乎无法识别到细胞，而在区域淋巴结水平可以检测到标记的细胞。结论：与其他动物模型相比，阴道HIV感染的hu-SCID小鼠模型具有简单和实用的特点，可能有助于测试化合物抗细胞相关HIV和其他性传播疾病的活性。(C) 2001利平科特·威廉姆斯·威尔金斯。
Objective: To develop an animal model of vaginal transmission of HIV-1 for the evaluation of vaginal microbicides.Design: Vaginal infection was performed in SCID mice reconstituted with 4 X 10(7) human peripheral blood lymphocytes (hu-PBL) by non-invasive vaginal administration. The hu-PBL were previously infected in vitro with a non-syncytium (NSI) strain of HIV-1 (SF162) (hu-PBL-SCID). Lymphocyte migration in vivo was examined using fluorescently labelled human lymphocytes.Methods: The percentage of CD4 T cells, plasma viral load and p24 antigen were evaluated using fluorescent activated cell sorting (FACS), the Amplicor HIV-1 monitor kit and enzyme-linked immunosorbent assay, respectively. Polymerase chain reaction (PCR) analysis was performed on DNA extracted from spleen and lymph nodes. For in vivo migration of labelled lymphocytes, the mice were sacrified after 4, 24 and 48 h; vaginae and local lymph nodes were removed, snap frozen with OCT, sectioned and examined by fluorescent microscopy and FACS.Results: HIV transmission was established using virus-infected cells inoculated vaginally, as shown by FACS, HIV viral load, p24 and PCR results. Labelled cells were easily located within the vaginal tissues after 4 h. However, few or no cells could be identified after 24 or 48 h at the vaginal level, whereas labelled cells could be detected at the level of regional lymph nodes.Conclusions: Because of its simplicity and practical features compared with other animal models, the vaginal HIV-infected hu-SCID mouse model may prove useful to test the activity of compounds against cell-associated HIV and, possibly, other sexually transmitted diseases. (C) 2001 Lippincott Williams Wilkins.