Pu Jeffrey J, Redman Colvin M, Visser Jan W M, Lee Soohee
Lindsley F. Kimball Research Institute of the New York Blood Center, New York, New York 10021, USA.
Transfusion. 2005 Jun;45(6):969-74. doi: 10.1111/j.1537-2995.2005.04289.x.
Kell and XK, two distinct red blood cell membrane proteins, are linked by a disulfide bond and form the Kell blood group complex. Kell surface antigens are expressed early during erythropoiesis but the onset of expression of XK which carries the Kx antigen is unknown.
To determine whether Kell and XK are synchronously expressed, sorted human hematopoietic progenitor cells and mouse progenitor cells of defined lineage were studied. To determine the onset of expression, human marrow and cord blood cells were sorted into three subpopulations, representing stem, multipotent, and erythroid progenitor cells, and the expression of Kell and XK was determined by reverse transcription-polymerase chain reaction (RT-PCR) and fluorescence-activated cell sorting (FACS) analysis. Mouse Kell and XK transcripts were determined by cDNA blotting of progenitor cells of defined lineage.
By RT-PCR, human peripheral blood progenitor cells had weak expression of Kell and XK transcripts but FACS analysis did not detect surface antigens. Kell and XK transcripts are expressed in multipotent progenitor cells and these cells express Kell surface antigens. The expression of Kx antigen in progenitor cells was not determined owing to nonspecific reactions with the antibody. By cDNA blotting, mouse Kell expression was detected in bipotential megakaryocytes-erythroid cells and in colony-forming units-erythroid (CFU-E) and burst-forming units-erythroid (BFU-E), whereas XK was only detected in CFU-E and BFU-E.
Both Kell and XK transcripts occur early during erythropoiesis; however, expression may not be coincident because, in mice, Kell transcripts, but not XK, occur in bipotential megakaryocytes-erythroid progenitor cells.
凯尔(Kell)蛋白和XK蛋白是两种不同的红细胞膜蛋白,它们通过二硫键相连,形成凯尔血型复合物。凯尔表面抗原在红细胞生成早期表达,但携带Kx抗原的XK蛋白的表达起始时间尚不清楚。
为了确定凯尔蛋白和XK蛋白是否同步表达,对分选的人造血祖细胞和特定谱系的小鼠祖细胞进行了研究。为了确定表达起始时间,将人骨髓和脐带血细胞分选成三个亚群,分别代表干细胞、多能祖细胞和红系祖细胞,并通过逆转录-聚合酶链反应(RT-PCR)和荧光激活细胞分选(FACS)分析来确定凯尔蛋白和XK蛋白的表达。通过对特定谱系祖细胞的cDNA印迹分析来确定小鼠凯尔蛋白和XK蛋白的转录本。
通过RT-PCR检测,人外周血祖细胞中凯尔蛋白和XK蛋白转录本表达较弱,但FACS分析未检测到表面抗原。凯尔蛋白和XK蛋白转录本在多能祖细胞中表达,且这些细胞表达凯尔表面抗原。由于抗体的非特异性反应,未确定祖细胞中Kx抗原的表达情况。通过cDNA印迹分析,在双潜能巨核细胞-红系细胞以及红系集落形成单位(CFU-E)和红系爆式集落形成单位(BFU-E)中检测到小鼠凯尔蛋白的表达,而XK蛋白仅在CFU-E和BFU-E中检测到。
凯尔蛋白和XK蛋白转录本均在红细胞生成早期出现;然而,表达可能不同步,因为在小鼠中,双潜能巨核细胞-红系祖细胞中存在凯尔蛋白转录本,但不存在XK蛋白转录本。