朗格汉斯细胞特异性递呈在巨噬细胞亲和性人类免疫缺陷病毒感染中的作用

The effect of specific virus delivering of Langerhans cell for the M-tropic HIV infection

目的 观察巨噬细胞亲和性HIV经皮肤感染的细胞学机制 ,区别巨噬细胞亲和性及T细胞亲和性HIV感染的差异。方法 以基因枪转染AD8及NL4 3分子克隆至皮肤 ,流式细胞仪和荧光显微镜观察病毒在表皮细胞中的表达 ,磁分离术分离CD80 + 和CD80 -的游离细胞 ,进而以PCR和逆转录酶检测游走细胞中HIV对CD+ 4 T淋巴细胞的感染和表达。结果 AD8及NL4 3经基因枪直接转染皮肤后 ,表皮游走细胞皆显示HIV转染 ,可扩增到gag基因。只有AD8转染的游走细胞能进一步感染CD4 + T淋巴细胞 ,CD4 + T淋巴细胞可扩增到gag并表达逆转录酶 ;含NL4 3的游走细胞 ,不能感染CD4 + T淋巴细胞 ,不能扩增到gag,不能表达逆转录酶。AD8转染的 1/ 2log稀释的CD80 + 细胞能够感染CD4 + T淋巴细胞。即使 4次稀释后 ,仍能感染CD4 + T细胞 ,PCR扩增到gag并有大量逆转录酶表达 ,超过对照组的 2～ 3倍。AD8转染的CD80 -细胞仅第 1次稀释与CD4 + T细胞共孵可扩增到gag,仍可见逆转录酶表达。进一步稀释后不能扩增到gag。NL4 3转染的CD80 + 或CD80 -游走细胞与CD4 + T细胞共孵 ,皆不能扩增到gag ,也无逆转录酶表达。结论 直接转染AD8的CD80 + 的游走表皮细胞能特异性地感染CD4 + T细胞。这种特异性不仅依赖于细胞表面HIV受体的亲和性结合 ,还依赖于转?

Objective To observe the cellular mechanism of via skin infection of Microphage tropic (M-tropic) HIV and further differentiate the infection of M-tropic and T-tropic HIV. Methods DNA clone of AD8 and NL43 was respectively transfected to skin by Gene Gun, The expression of virus in the epidermal cells was observed by flow-cytometry and fluoroscope. Emigrating cells were Mac-sorted into CD80 + and CD80 -. The infection of CD4 +T lymphocytes by HIV from emigrating cells were further assessed by PCR and RT assay. Results Through direct transfection of skin by Gene Gun, emigrating cells from skin showed transfection of AD8 and NL43 and the gag gene can be amplified from the cells. Only AD8 transfected emigrating cells can further infect CD4 +T lymphocyte. Gag can be amplified and reverse transcriptase can be expressed from these CD4 + T cells; NL43 transfected emigrating cells can not infect CD4 +T lymphocyte. No gag can be amplified and no reverse transcriptase can be expressed. AD8 transfected and 1/2 log diluted CD80 + cells can infect CD4 +T lymphocyte. Even after 4 dilutions, There are still a lot of CD4 +T cells,gag can be amplified and a large amount of reverse transcriptase was expressed that is double or triple of controls. AD8 infected CD80 -cells co-cultured with CD4 +Tcells showed gag amplification only in first dilution and reverse transcriptase is also expressed. Gag can not be amplified by further dilution. NL43infected CD80 + or CD80 - emigrating cells co-cultured with CD4 +T cells did not show gag amplification and did not express reverse transcriptase. Conclusion Direct AD8 transfected CD80 + emigrating epidermal cells can specifically infect CD4 +T lymphocyte. This specificity not only depends on affinitive combination of HIV receptor on cell surface, but also on the specific virus delivering of AD8 transfected CD80 + cells that infect the CD4 +Tcells. That means even without the receptor, CD80 + cells only deliver transfected AD8 but not NL43. So, It can be said that the specificity of the virus receptor is not the only clue of HIV infection.