Use of the fluorescence-activated cell sorter to quantitate and enrich for subpopulations of human skin cells.
Use of the fluorescence-activated cell sorter to quantitate and enrich for subpopulations of human skin cells.
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使用荧光激活细胞分选仪对人类皮肤细胞亚群进行定量和富集。
DOI：
10.1111/1523-1747.ep12500077
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发表时间：
1982
期刊：
影响因子：
--
通讯作者：
Engleman,EG
中科院分区：
文献类型：
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作者：
Morhenn,VB;Benike,CJ;Charron,DJ;Cox,A;Mahrle,G;Wood,GS;Engleman,EG
在人表皮细胞抗原表达的定量研究中，比较了各种免疫染色技术。几乎所有的有核表皮细胞都表达β2-微球蛋白，它与HLA-A、-B和-C抗原相关，而只有约4%表达T6，这是一种由朗格汉斯细胞表达的抗原，而不是皮肤中的其他细胞。用流式细胞仪（FACS）对表皮细胞悬液中的T6阳性朗格汉斯细胞进行了10 ~ 15倍的选择性富集，平均6.5%的细胞被抗HLA-DR抗体特异性染色。当分散的细胞与抗DR加过氧化物酶染色的免疫电镜技术进行了检查，只有单核白细胞（可能朗格汉斯细胞）染色。在用FACS分离HLA-DR阳性皮肤细胞后，DR阳性群体而不是DR阴性群体刺激同种异体应答淋巴细胞的增殖，表明分选的细胞具有代谢活性。我们的结论是，HLA-DR抗原不表达角质形成细胞在正常的人皮肤细胞悬浮液，流式细胞仪可用于选择性地丰富或耗尽皮肤细胞悬浮液的抗原不同的亚群，如朗格汉斯细胞。
A variety of immunologic staining techniques were compared in a quantitative study of antigen expression by human epidermal cells. Virtually all nucleated epidermal cells expressβ2-microglobulin, which is associated with HLA-A, -B, and -C antigens, whereas only about 4% expressed T6, an antigen expressed by Langerhans cells but not other cells in the skin. With the fluorescence-activated cell sorter (FACS), epidermal cell suspensions were selectively enriched 10- to 15-fold for T6-positive Langerhans cells.An average of 6.5% of cells were specifically stained by anti-HLA-DR antibody. When dispersed cells stained with anti-DR plus peroxidase were examined with the technique of immunoelectron microscopy, only mononuclear leukocytes (probably Langerhans cells) were stained. After separating HLA-DR positive skin cells with the FACS, the DR-positive population but not the DR-negative population stimulated proliferation of allogeneic responder lymphocytes, indicating that sorted cells are metabolically active. We conclude that HLA-DR antigen is not expressed by keratinocytes in normal human skin cell suspensions and that the FACS can be used to selectively enrich or deplete skin cell suspensions of antigenically distinct subpopulations such as Langerhans cells.