Peters P M, Kamarck M E, Hemler M E, Strominger J L, Ruddle F H
J Exp Med. 1984 May 1;159(5):1441-54. doi: 10.1084/jem.159.5.1441.
The genes that code for the human lymphocyte cell surface determinants defined by monoclonal antibodies A- 1A5 and A- 3A4 have been genetically mapped. All human chromosomes, except Y, were included in a series of human less than mouse lymphocyte hybrid populations that retained expression of lymphocyte-specific surface markers. Expression of the A- 1A5 and A- 3A4 antigens was quantitated by indirect immunofluorescence and fluorescence-activated cell sorter (FACS) analysis. Hybrid populations heterogeneous for antigen expression were sorted to yield antigenically homogeneous subpopulations. Isozyme analysis indicated concordant segregation of the A- 1A5 determinant with chromosome 10, and the A- 3A4 determinant with chromosome 4. In contrast to the unhybridized human parent cell line (MOLT-4), from which A- 1A5 immunoprecipitated two proteins (160,000 and 125,000 Mr), A- 1A5 only immunoprecipitated a single band (125,000 Mr) from an A- 1A5 -expressing human less than mouse hybrid. The genetic disassociation of these two proteins from the A- 1A5 -reactive complex suggests that the appearance of the 160,000 Mr protein requires a gene locus that is unlinked to the locus for the 125,000 Mr protein on chromosome 10. A third component of the A- 1A5 -reactive protein complex (210,000 Mr), which is recognized by the monoclonal antibody TS2/7, was not expressed on the parent MOLT-4 cells, but was weakly expressed on MOLT-4 less than mouse BW5147 hybrids. This allowed preliminary mapping of that determinant to either chromosome 10 or 15. The A- 3A4 antigen (approximately 45,000 Mr) is a novel cell surface structure expressed on all hematopoietic cell lines tested, and represents the first cell surface marker mapped to chromosome 4.
编码由单克隆抗体A-1A5和A-3A4所定义的人类淋巴细胞细胞表面决定簇的基因已被进行了基因定位。除Y染色体外的所有人类染色体都包含在一系列人-小鼠淋巴细胞杂交群体中,这些杂交群体保留了淋巴细胞特异性表面标志物的表达。通过间接免疫荧光和荧光激活细胞分选仪(FACS)分析对A-1A5和A-3A4抗原的表达进行定量。对抗原表达异质性的杂交群体进行分选,以产生抗原性均一的亚群体。同工酶分析表明,A-1A5决定簇与10号染色体共分离,A-3A4决定簇与4号染色体共分离。与未杂交的人类亲本细胞系(MOLT-4)不同,从MOLT-4细胞中A-1A5可免疫沉淀出两种蛋白质(分子量分别为160,000和125,000),而从表达A-1A5的人-小鼠杂交细胞中A-1A5仅免疫沉淀出一条带(分子量125,000)。这两种蛋白质与A-1A5反应复合物的基因解离表明,分子量160,000蛋白质的出现需要一个与10号染色体上分子量125,000蛋白质的基因座不连锁的基因座。单克隆抗体TS2/7识别的A-1A5反应蛋白复合物的第三个成分(分子量210,000)在亲本MOLT-4细胞上不表达,但在MOLT-4-小鼠BW5147杂交细胞上弱表达。这使得该决定簇初步定位于10号或15号染色体。A-3A4抗原(约45,000分子量)是在所有测试的造血细胞系上表达的一种新型细胞表面结构,是第一个定位于4号染色体的细胞表面标志物。