人表皮细胞培养中Langerhans细胞变化的研究

研究表明表皮Langerhans细胞(Lcs)是一种重要的抗原递呈细胞,其参与同种皮肤排斥反应。如消除移植物内Lcs,可延长异体皮肤存活。本研究系利用ATPase染色,抗人HLA-Dr抗原单克隆抗体间接免疫萤光及免疫酶标(ABC)染色,观察了人表皮细胞(Ecs)培养中Lcs的密度及形态变化。并通过混合表皮细胞淋巴细胞培养反应(MEcLR),检查了培养前后Ecs的免疫刺激能力。发现在Ecs培养过程中,存于Ecs内的Lcs培养3天即显著减少同时明显变形;7天后Lcs减少达90％以上;培养14天或更长时间Ecs内Lcs基本消失。在培养新生的Ecs内自始未发现Lcs。培养7天后的Ecs及新生Ecs在MEcLR 中失去刺激异体淋巴细胞转化的能力。这些结果表明通过Ec体外培养可以消除Ecs内携带Ⅰ级抗原的Lcs,提示本培养系统培养的同种人Ecs如用于异体移植,其存活的时间可能延长。

Study of the Changes of Langerhans in Cultured Human Epidermal Cells

It has been shown that epidermal Langerhans cells (LCs) were antigen presenting cells in cutaneous cell-mediated immuneresponse,and they were implicated in skin allograft rejection,If LCs can be depleted from skin grafts,the survival period of the allografts may be prolonged. Therefore,we observed the changes of LCs in cultured human epidermal cells (ECs) by ATPase staining,HLA-Dr monoclonal antibody indirect immunofluoresence and enzyme labelling avidin-biotin complex (ABC) staining.And the capacity of ECs to stimulate allogeneic lymphocyte proligeration before and after cultures was testedby mixed epidermal cell-lymphocyte culture response (MEcLR).We found that LCs in original skin explants were significantly decreased and deformed three days after culture.LCs were reduced more than 90 percent after 7 days of culture.LCs in original skin explants were approximately eliminated 14 days or more of culture.LCs were never seen within epidermal outgrowth in culture.The Ecs gained from the original skin explants which were cultured after 7 days and from epidermal outgrowth lost their ability to stimulate allogeneic lymphocyte transformation in MEcLR.These results indicate that LCs bearing HLA-Dr antigen in human are able to be depleted from epidermal grafts by epidermal cell culture,and suggest that the ECs cultured in our culture system can prolong their survival period as they are used as allografts.