Co-expression of CXCR4/fusin and galactosylceramide in the human intestinal epithelial cell line HT-29
Co-expression of CXCR4/fusin and galactosylceramide in the human intestinal epithelial cell line HT-29
复制标题
DOI：
10.1097/00002030-199711000-00004
复制
发表时间：
1997-09-01
期刊：
影响因子：
3.8
通讯作者：
Fantini, J
中科院分区：
文献类型：
--
作者：
Delezay, O;Koch, N;Fantini, J
目的：检测CXCR 4/fusin在人肠上皮细胞中的表达，探讨其在gp 120受体半乳糖神经酰胺（galactosylceramide，CalCer）介导的HIV-1感染途径中的作用。CalCer+（HT-29，HT-29/CD 4+）和GalCer-使用单克隆抗体（MAb）12 C5分析（Caco-2/C12、C114和C114/CD 4+）人肠细胞系的CXCR 4/融合蛋白表达。然后评价这种单克隆抗体在允许细胞中抑制HIV-1感染的能力。结果：HT-29和HT-29/CD 4+细胞表面可检测到CXCR 4/fusin，而Caco-2/Cl 2、C/14和Cl 74/CD 4+细胞表面均未检测到CXCR 4/fusin。90%的CXCR 4/融合蛋白+ HT-29和HT-29/Cd 4+细胞共表达GalCer。HIV-1实验室分离株感染HT-29细胞可被抗GalCer和抗CXCR 4/融合素单克隆抗体抑制。CD 4的表达使HT-29细胞对HIV-1（89.6）敏感，HIV-1是一种不识别GalCer的嗜巨噬细胞分离株。12 G5单抗阻断HIV-1感染HT-29/CD 4+细胞。相反，HIV-1受体的表达，即，CD 4，CalCer或两者，进入CXCR 4/融合蛋白阴性肠细胞并不赋予对HIV-1感染的敏感性。然而，由此产生的受体阳性细胞系可以结合HIV-1，而原来的细胞系不能。结论：HIV-1进入人肠细胞涉及GalCer和CXCR 4/fusin。HIV-1分离株如89.6能够使用CXCR 4/融合素作为辅助受体，但不与CalCer结合，不感染这些细胞。这些数据提出了CXCR 4/融合蛋白可能作为HIV-1进入CD 4-/GalCer+肠上皮细胞的辅助受体的可能性。
Objective: To detect the expression CXCR4/fusin in human intestinal epithelial cells and to assess its potential role in the pathway of HIV-1 infection mediated by the alternative gp120 receptor galactosylceramide (CalCer).Methods: CalCer+ (HT-29, HT-29/CD4+) and GalCer-(Caco-2/Cl2, Cl14 and C114/CD4+) human intestinal cell lines were analysed for CXCR4/fusin expression using the monoclonal antibody (MAb) 12C5. This MAb was then evaluated for its ability to inhibit HIV-1 infection in permissive cells. HIV-1 infection was measured by detection of p24 antigen, polymerase chain reaction amplification, and cocultivation with CD4+ cells.Results: CXCR4/fusin was detected on the surface of HT-29 and HT-29/CD4+, but not on Caco-2/Cl2, C/14 and Cl74/CD4+ cells. Ninety per cent of CXCR4/fusin+ HT-29 and HT-29/Cd4+ cells co-expressed GalCer. Infection of HT-29 cells by laboratory isolates of HIV-l was inhibited by both anti-GalCer and anti-CXCR4/fusin MAbs. Expression of CD4 rendered HT-29 cells sensitive to HIV-1(89.6), a macrophage-tropic isolate that does not recognize GalCer. The 12G5 MAb blocked HIV-1 infection of HT-29/CD4+ cells. In contrast, the expression of HIV-1 receptors, i.e., CD4, CalCer or both, into CXCR4/fusin-negative intestinal cells did not confer sensitivity to HIV-1 infection. The resulting receptor-positive cell lines could, however, bind HIV-1, whereas the original cell lines could not.Conclusion: HIV-1 entry into human intestinal cells involves both GalCer and CXCR4/fusin. HIV-1 isolates such as 89.6 that are able to use CXCR4/fusin as coreceptor, but do not bind to CalCer, do not infect these cells. These data raise the possibility that CXCR4/fusin may function as a coreceptor for HIV-1 entry into CD4-/GalCer+ intestinal epithelial cells.