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A human leukocyte antigen identified by a monoclonal antibody
Authors:Ann E. Berger  Janet E. Davis  Peter Cresswell
Affiliation:Division of Immunology, Duke University of Medical Center, Durham, NorhtCarolina, USA
Abstract:Leukocyte antigen of approximate molecular weight 200,000 daltons have beendescribed in mouse, rat, and man. We described here the reactivities of a monoclonal antibody. GAP 8.3, which identified such an antigen on human leukocytes. We found the leukocyte antigen H-T200 on T and B lymphocytes, granulocytes, monocytes, and platelets, but not on erythrocytes or nonhematopoetically derived cells. Resting and activated T cells had more antigen or their surfaces than did resting B lymphocytes and EBV-transformed B cells, respectively. The leukocyte antigen was detected on approximately 75% of bone marrow cells; cells of the erythroid series comprised the negative population. The GAP 8.3 antibody and its F(ab′)2 fragments had no effect on in vitro stimulation of peripheral blood cells by mitogens or allogeneic cells. Antigen isolated from T cell lines had a higher electrophoretic mobility than did antigen from B cell lines; antigen from the mycloid line U937 comigrated with that from B cell lines. In addition, we detected very small but reproducible differences in the electrophoretic mobility of the antigen on two T cell lines.
Keywords:LCL  lymphoblastoid cell line  L/C  leukocyte common  H-T200  human T200  PHA  phytohemagglutinin  PWM  pokeweed mitogen  Con A  concanavalin A  PBS  phosphate buffered saline  HAT  hypoxanthine, amethopterin, thymidine  FCS  fetal calf (bovine) serum  LSM  lymphocyte separating medium  PBA  PBS containing 2% bovine, serum albumin and 0.02%, sodium azide  TBS  0.01 M TRIS, 0.15 M NaCl, pH 8.2  octa-ethyleneglycol dodecyl, ether  CYNAP  cytotoxicity negative, absorption positive  A/A  antibiotic/antimucotic
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