Generation and characterization of a monoclonal antibody that recognizes an activation antigen expressed by a majority of adherent lymphokine-activated killer cells.
In an attempt to generate murine natural killer (NK) cell-specific monoclonal antibodies (MAbs) by immunizing Balb/c mice with C57BL/6 (B6) A-LAKs, we have isolated a hybridoma, CS/NicT.2, which secretes an IgM that recognizes a majority of B6 and B6 Rag-1-/- A-LAKs. The CS/NicT.2 antigen is highly expressed by A-LAKs, but only at extremely low levels on resting splenocytes, suggesting that its expression is tightly associated with IL-2 activation. Among the cell lines examined, only CTLL-2 expresses the CS/NicT.2 antigen at relatively high levels. A low level of CS/NicT.2 staining is also detected on resting allo-specific cytotoxic T lymphocytes (CTL) clones, AB.1 and C11. In addition, a similar low level of CS/NicT.2 staining is detected on the T-helper cell line HT-2. The CS/NicT.2 antigen is upregulated by ionomycin but not by phorbol 12-myristate 13-acetate (PMA). For the CTL clones examined, CS/NicT.2 staining is also dramatically increased by anti-TCRbeta or anti-CD3epsilon stimulation. Protease treatments of CTLL-2 show that this antigen is proteinase K sensitive, but relatively resistant to trypsin digestion. Furthermore, the CS/NicT.2 antigen exhibits a relatively fast turnover rate as assessed by proteinase K and cycloheximide treatments of CTLL-2, suggesting that the CS/NicT.2 antigen may have a short half-life on the cell surface.
Chang
C S
CS
Department of Medical Microbiology and Immunology, Faculty of Medicine, University of Alberta, Edmonton, Canada.
Kane
K P
KP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000378:metabolism; D000943:Antigens, Differentiation / Q000096:biosynthesis*; D002448:Cell Adhesion / Q000276:immunology; D010450:Endopeptidases / Q000378:metabolism; D005260:Female; D015759:Ionomycin / Q000494:pharmacology; D015979:Killer Cells, Lymphokine-Activated / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology / Q000378:metabolism; D015854:Up-Regulation / Q000187:drug effects / Q000276:immunology
D000911:Antibodies, Monoclonal; D000943:Antigens, Differentiation; D015759:Ionomycin; D010450:Endopeptidases