A Comparative Analysis of Cell Surface Markers on Murine NK Cells and CTL Target-Effector Conjugates |
| |
| Authors: | T.J. BEAUMONT J. C. RODER B. E. ELLIOTT R. S. KERBEL J.W. DENNIS M. KASAI K. OKUMURA |
| |
| Affiliation: | Department of Microbiology and Immunology, and The Cancer Research Division, Department of Pathology, Queen's University, Kingston, Ontario, Canada;Institute of Biology Sciences, University of Tsukuba, Ibaraki, and Department of Immunology, Faculty of Medicine, University of Tokyo, Tokyo, Japan |
| |
| Abstract: | The rosetting of sheep erythrocytes (SRBC) coated with non-haemagglutinating monoclonal antibodies rather than conventional haemagglutinating antisera revealed readily detectable FcR on most splenic natural killer (NK) cells since 76% of splenic lymphocytes forming conjugates with YAC also resetted with SRBC coated with high concentrations of monoclonal anti-SRBC antibody of the IgG2b subclass and since Ficoll depletion or enrichment of splenic lymphocytes rosetting with IgG2b-coated SRBC resulted in a corresponding 4-fold decrease or increase in conjugate-forming cells and a 10-fold decrease or increase in NK cytolytic acttvity. NK cells bound much less readily to monoclonal IgG2a and not at all to monoclonal IgGI or IgM, but the degree of binding was directly proportional to the amount of antibody on the erythrocytes and was not isotype-restricted. In addition, immunofluorescent studies revealed that YAC-1-conjugated lymphocytes were Lyt-1-, Lyt-2-, partially Thy-1+ (60%), asiato-GMI + (80%), Qa-4+ (77%), Qa-5+ (79%), and Ly-5+ (94%). In comparison, a proportion (39%) of alloimmune peritoneal exudate cells which conjugated with P815–2 also siained by immunofluorescence with anti-asialo GM1 antisera. Most (>90%) P815- conjugated cells were Thy-1+, Lyt-2+. and a subpopulation of Lyt-l+2+ conjugates was observed (25 %). Qa-5 and Ly-5 were also expressed on most (two-thirds) cytolytic T lymphocytes (CTL) conjugates, whereas Qa-4 and FcR for IgG2b were not detected. The best phenotypic distinctions between NK cells and CTL were therefore based on the presence or absence of Lyt-2, Qa-4, and FcR for IgG2b on most effector cells. Anti-asialo-GMl or monoclonal anti-Qa-4 and complement treatment greatly diminished both the frequency of NK conjugates and the percentage of conjugates with detectable IgG2b FcR or asialo-GM1. These results confirm that NK cells co-express asialo-GMI and Fc receptors, at the single-celt level, and provide a simple method for greatly enriching NK populations at least 10-fold. |
| |
| Keywords: | |
|
|
