Ottmann O G, Nocka K H, Moore M A, Pelus L M
Department of Hematopoietic Regulation, Sloan-Kettering Institute, New York, NY 10021.
Leukemia. 1988 Oct;2(10):677-86.
Expression of major histocompatibility complex class II Ags HLA-DR, HLA-DP, and HLA-DQ on human BM granulocyte-erythroid-macrophage-megakaryocyte CFU (CFU-GEMM), BFU-E, and CFU-GM was examined by indirect immunofluorescence, cell sorting, and complement-mediated cytotoxicity. BM, highly enriched for progenitor cells by depletion of mature hematopoietic elements, was further separated by sterile sorting into HLA-DR (-), low, intermediate, and high intensity HLA-DR (+), as well as HLA-DP (+) and HLA-DP (-) cell fractions and assayed for progenitor cell content. In addition, in the case of HLA-DR, CFU-GM response to inhibition by prostaglandin E was determined. Cell sorting and cytotoxicity data confirm that approximately 95% of assayable erythroid, myeloid, and multipotential progenitor cells expressed HLA-DR, whereas HLA-DQ Ags were undetectable. HLA-DR and HLA-DP Ags were co-expressed on 61% of these progenitor cells, predominantly those expressing HLA-DR at high intensity. Day 7 and 14 CFU-GM showed a trend toward segregation to the high HLA-DR (+) cell fractions, especially when recombinant human G-CSF was used to stimulate clone formation. Both day 7 and day 14 CFU-GMs were found predominantly in the HLA-DP (+) cell fraction. In contrast, BFU-E and CFU-GEMM were found in the low intensity HLA-DR cell fraction and predominantly in the HLA-DP (-) fraction. Both eosinophil CFU and cells giving rise to basophil/mast cells in suspension culture were found in the low and intermediate intensity HLA-DR fractions, but could be segregated into HLA-DP (+) and HLA-DP (-) cell fractions, respectively. Functional analysis of day 7 CFU-GM segregated, based upon HLA-DR intensity, indicated a positive correlation between increasing HLA-DR intensity and responsiveness to inhibition by prostaglandin E. Furthermore, only those CFU-GM expressing HLA-DR at high intensity could be removed by cytolytic treatment using a mAb anti-HLA-DR previously shown to be selective for CFU-GM responsive to PGE and in S phase of the cell cycle.
通过间接免疫荧光、细胞分选和补体介导的细胞毒性检测人骨髓粒系-红系-巨噬系-巨核系集落形成单位（CFU-GEMM）、爆式红系集落形成单位（BFU-E）和粒系集落形成单位（CFU-GM）上主要组织相容性复合体II类抗原HLA-DR、HLA-DP和HLA-DQ的表达。通过去除成熟造血细胞成分高度富集祖细胞的骨髓，再经无菌分选进一步分为HLA-DR(-)、低强度、中等强度和高强度HLA-DR(+)，以及HLA-DP(+)和HLA-DP(-)细胞组分，并检测祖细胞含量。此外，对于HLA-DR，测定CFU-GM对前列腺素E抑制的反应。细胞分选和细胞毒性数据证实，约95%可检测的红系、髓系和多能祖细胞表达HLA-DR，而未检测到HLA-DQ抗原。HLA-DR和HLA-DP抗原在61%的这些祖细胞上共表达，主要是那些高强度表达HLA-DR的细胞。第7天和第14天的CFU-GM显示出向高HLA-DR(+)细胞组分分离的趋势，特别是当使用重组人粒细胞集落刺激因子（G-CSF）刺激克隆形成时。第7天和第14天的CFU-GM主要存在于HLA-DP(+)细胞组分中。相反，BFU-E和CFU-GEMM存在于低强度HLA-DR细胞组分中，且主要存在于HLA-DP(-)组分中。悬浮培养中嗜酸性粒细胞集落形成单位和产生嗜碱性粒细胞/肥大细胞的细胞均存在于低强度和中等强度HLA-DR组分中，但可分别分离到HLA-DP(+)和HLA-DP(-)细胞组分中。基于HLA-DR强度对第7天CFU-GM进行分离的功能分析表明，HLA-DR强度增加与对前列腺素E抑制的反应性呈正相关。此外，只有那些高强度表达HLA-DR的CFU-GM才能通过使用先前显示对前列腺素E有反应且处于细胞周期S期的抗HLA-DR单克隆抗体进行溶细胞处理而被去除。
