PRODUCTION OF MONOCLONAL ANTIBODIES TO GROUP-A ERYTHROCYTES, HLA AND OTHER HUMAN CELL-SURFACE ANTIGENS - NEW TOOLS FOR GENETIC-ANALYSIS
PRODUCTION OF MONOCLONAL ANTIBODIES TO GROUP-A ERYTHROCYTES, HLA AND OTHER HUMAN CELL-SURFACE ANTIGENS - NEW TOOLS FOR GENETIC-ANALYSIS
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DOI：
10.1016/0092-8674(78)90296-9
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发表时间：
1978-01-01
期刊：
影响因子：
64.5
通讯作者：
ZIEGLER, A
中科院分区：
文献类型：
--
作者：
BARNSTABLE, CJ;BODMER, WF;ZIEGLER, A
分泌抗体的杂交细胞来源于用人扁桃体淋巴细胞制备的膜免疫的小鼠骨髓瘤细胞和脾脏细胞的融合。通过培养上清检测与人扁桃体细胞结合的抗体，检测对细胞表面抗原分泌抗体的杂交体。分析了6种不同的抗体（W6/1、/28、/32、/34、/45和/46）。这些抗原的组织分布广泛（W6/32、/34、/45和/46），或主要作用于红细胞（W6/1和W6/28）。其中一种抗红细胞抗体（W6/1）检测到多态性抗原，因为A1和A2血型的红细胞被标记，而B和O血型的红细胞没有被标记。抗体W6/34、/45和/46均针对小鼠-人杂交种11号染色体短臂上的抗原。免疫沉淀研究表明，W6/45抗原可能是16000道尔顿表观分子量的蛋白，而W6/34和/46抗原不能被该技术检测到。抗体W6/32针对的是HLA-A、B和C抗原的43,000道尔顿分子量链中大多数（如果不是全部）共有的决定因素。这是通过体细胞遗传技术和免疫沉淀分析建立的。扁桃体白细胞在饱和状态下每个细胞结合370,000个W6/32抗体分子。克隆了杂交骨髓瘤W6/32和W6/34。两者都分泌一种免疫球蛋白G2抗体。在小鼠体内培养W6/32细胞，荷瘤动物血清中含有bb0 - 10 mg/ml单克隆抗体。实验证实了混合骨髓瘤技术在制备抗人细胞表面抗原的单特异性抗体方面的有效性。强调了获得体细胞遗传学试剂和获得检测人类抗原多态性的小鼠抗体的可能性。
Antibody-secreting hybrid cells were derived from a fusion between mouse myeloma cells and spleen cells from a mouse immunized with membrane from human tonsil lymphocyte preparations. Hybrids secreting antibodies to cell surface antigens were detected by assaying culture supernatants for antibody binding to human tonsil cells. Six different antibodies (called W6/1,/28,/32,/34,/45 and/46) were analyzed. These were against antigens of wide tissue distribution (W6/32, /34, /45 and /46) or mainly on erythrocytes (W6/1 and W6/28). One of the anti-erythrocyte antibodies (W6/1) detected a polymorphic antigen, since blood group A1 and A2 erythrocytes were labeled while B and O were not. Antibodies W6/34, /45 and /46 were all against antigens which were mapped to the short arm of chromosome 11 by segregation analysis of mouse-human hybrids. Immunoprecipitation studies suggest that W6/45 antigen may be a protein of 16,000 dalton apparent MW while W6/34 and /46 antigens could not be detected by this technique. Antibody W6/32 is against a determinant common to most, if not all, of the 43,000 dalton MW chains of HLA-A, B and C antignes. This was established by somatic cell genetic techniques and by immunoprecipitation analysis. Tonsil leukocytes bound 370,000 W6/32 antibody molecules per cell at saturation. The hybrid myelomas W6/32 and W6/34 were cloned. Both secrete an Ig[immunoglobulin]G2 antibody. W6/32 cells were grown in mice and the serum of the tumor-bearing animals contained > 10 mg/ml of monoclonal antibody. The experiments established the usefulness of the hybrid myeloma technique in preparing monospecific antibodies against human cell surface antigens. The possibilities of obtaining reagents for somatic cell genetics and of obtaining mouse antibodies detecting human antigenic polymorphisms were highlighted.