Mechanism of target cell recognition by CD3- LGL. I. Development of a monoclonal antibody to a K562-associated target cell antigen.
In an attempt to identify the target recognition molecule(s) involved in the interaction between CD3- large granular lymphocyte (LGL) and a tumor cell target, monoclonal antibodies (mAb) to NK-susceptible K562 tumor cell membrane glycoproteins were developed. After screening by ELISA for reactivity to K562 membrane glycoproteins, two monoclonal antibodies were identified (mAb 35 and mAb 36). One of the monoclonal antibodies (mAb 36) was found to inhibit conjugation between LGL and K562 target cells and also to inhibit lysis of K562 by LGL. Upon further testing, mAb 36 also inhibited the binding between LGL and other NK-susceptible target cells, e.g., Daudi and Molt 4. In contrast, mAb 35, even though binding to K562, did not inhibit the binding of LGL to tumor targets and therefore was used as an isotype control. When mAb 36 was utilized as an affinity matrix, bound proteins specifically inhibited CD3- LGL-K562 conjugation. Experiments involving tunicamycin treatment of tumor target cells demonstrated that mAb 36 recognized a carbohydrate moiety rather than the protein core. Therefore, these data suggested that the target cell recognition molecule which is recognized by mAb 36 appears to be a membrane carbohydrate-associated molecule.
Bino
T
T
Israel Institute for Biological Research, Ness-Ziona.
Frey
J L
JL
Ortaldo
J R
JR
D000911:Antibodies, Monoclonal / Q000276:immunology*; D000936:Antigen-Antibody Complex / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology* / Q000302:isolation & purification; D002460:Cell Line; D002846:Chromatography, Affinity; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007075:Immunoglobulin M / Q000276:immunology*; D007694:Killer Cells, Natural / Q000276:immunology*; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000276:immunology / Q000473:pathology; D008562:Membrane Glycoproteins / Q000276:immunology* / Q000302:isolation & purification; D014415:Tunicamycin / Q000494:pharmacology
D000911:Antibodies, Monoclonal; D000936:Antigen-Antibody Complex; D000954:Antigens, Surface; D007075:Immunoglobulin M; D008562:Membrane Glycoproteins; D014415:Tunicamycin