Harrington R D, Geballe A P
Department of Molecular Medicine, Fred Hutchinson Cancer Research Center, Seattle, WA 98104, USA.
Ann Clin Lab Sci. 1996 Nov-Dec;26(6):522-30.
Expression of CD4, the principle receptor for HIV-1, is not sufficient for viral entry into most non-human and rare human cell lines. Construction of HIV-1 susceptible heterokaryons and a hybrid cone by fusion of HeLa cells and the HIV-1 resistant human cell line U373-CD4 were previously reported by us. These results suggested that U373-CD4 lack a cofactor(s) which is essential for HIV-1 entry and can be supplied by HeLa cells. Now the construction of multiple stable U373-CD4/HeLa whole cell and microcell hybrid clones are described, two of which are highly susceptible to HIV-1. Using these hybrids it is demonstrated that expression of CD4 and CD26, the T cell activation antigen dipeptidyl peptidase IV recently proposed as a CD4 cofactor necessary for HIV-1 infection, are not sufficient for HIV-1 entry. This panel of HIV-1 resistant and susceptible hybrids provides a rapid, simple assay for testing the role of other candidate cofactor molecules (e.g., fusin) that may be required for HIV-1 entry into human cells. Further, the method described by us for constructing HeLa microcells should permit the construction of murine-HeLa microcell hybrids, thus providing ideal reagents for determining which human chromosome(s) are needed to confer HIV-1 susceptibility onto non-human cells.
CD4作为HIV-1的主要受体，其表达并不足以使病毒进入大多数非人类及少数人类细胞系。我们之前报道过通过HeLa细胞与对HIV-1耐药的人类细胞系U373-CD4融合构建对HIV-1敏感的异核体和杂交细胞。这些结果表明U373-CD4缺乏一种对HIV-1进入至关重要且可由HeLa细胞提供的辅助因子。现在描述了多个稳定的U373-CD4/HeLa全细胞和微细胞杂交克隆的构建，其中两个对HIV-1高度敏感。利用这些杂交细胞证明，CD4和CD26（最近被认为是HIV-1感染所需的CD4辅助因子的T细胞活化抗原二肽基肽酶IV）的表达并不足以使HIV-1进入。这组对HIV-1耐药和敏感的杂交细胞为测试其他可能是HIV-1进入人类细胞所需的候选辅助因子分子（如融合素）的作用提供了一种快速、简单的检测方法。此外，我们描述的构建HeLa微细胞的方法应该能够构建鼠-HeLa微细胞杂交体，从而为确定将HIV-1易感性赋予非人类细胞需要哪些人类染色体提供理想的试剂。
