Stroncek D F, Plachta L B, Herr G P, Dalmasso A P
University of Minnesota, Minneapolis.
Transfusion. 1993 Aug;33(8):656-60. doi: 10.1046/j.1537-2995.1993.33893342747.x.
The neutrophil-specific NB antigen system has been serologically characterized with human alloantisera. Two alleles, NB1 and NB2, have been described. NB1 is expressed on a subpopulation of peripheral blood neutrophils in 97 percent of healthy donors. Human alloantibodies have been used to identify the 58- to 64-kDa glycoprotein (GP) on which NB1 is located. NB1 can usually be detected by both a granulocyte immunofluorescence (GIF) assay and a granulocyte agglutination (GA) assay, but neutrophils from some donors have been found to react with anti-NB1 in GIF but not in GA assays. To determine if the latter neutrophils express NB1 and the corresponding 58- to 64-kDa GP, these neutrophils were probed with rabbit and human sera specific for NB1. First, the proportion of neutrophils that express NB1 was quantitated. Neutrophils from donors that typed as NB1-positive in both GA and GIF assays were analyzed by flow cytometry with antisera to NB1. Human and rabbit anti-NB1 reacted with 71 +/- 17 percent and 70 +/- 17 percent of neutrophils, respectively. There was no difference in the expression of NB1 in NB1-homozygous and NB1-heterozygous individuals. In contrast, significantly fewer neutrophils from four donors that typed as NB1-positive in GIF assay but not GA assay reacted with human (27 +/- 12%; p < 0.001) and rabbit (26 +/- 12%; p < 0.001) anti-NB1. When neutrophils from these same four donors were probed with rabbit and human anti-NB1 by immunoblotting and immunoprecipitation, the 58- to 64-kDa GP was identified.(ABSTRACT TRUNCATED AT 250 WORDS)
中性粒细胞特异性NB抗原系统已通过人同种异体抗血清进行了血清学特征分析。已描述了两个等位基因，即NB1和NB2。NB1在97%的健康供体的外周血中性粒细胞亚群上表达。人同种异体抗体已用于鉴定NB1所在的58至64 kDa糖蛋白（GP）。NB1通常可通过粒细胞免疫荧光（GIF）测定法和粒细胞凝集（GA）测定法检测到，但已发现一些供体的中性粒细胞在GIF测定中与抗NB1反应，而在GA测定中不反应。为了确定后一种中性粒细胞是否表达NB1和相应的58至64 kDa GP，用对NB1特异的兔血清和人血清对这些中性粒细胞进行检测。首先，对表达NB1的中性粒细胞比例进行定量。对在GA和GIF测定中均为NB1阳性的供体的中性粒细胞，用抗NB1抗血清通过流式细胞术进行分析。人抗NB1和兔抗NB1分别与71±17%和70±17%的中性粒细胞反应。NB1纯合子和NB1杂合子个体中NB1的表达没有差异。相比之下，在GIF测定中为NB1阳性但在GA测定中不是的四名供体的中性粒细胞与抗人（27±12%；p<0.001）和抗兔（26±12%；p<0.001）抗NB1反应的明显较少。当用免疫印迹法和免疫沉淀法用兔抗NB1和人抗NB1对来自这相同四名供体的中性粒细胞进行检测时，鉴定出了58至64 kDa的GP。（摘要截断于250字）
