Immunoelectron microscopic identification of human NK cells by FITC-conjugated anti-Leu-11a and biotinylated anti-Leu-7 antibodies.

摘要: Abstract Human natural killer (NK) cells have been reported to express various surface antigens. The majority and the most functionally potent NK are Leu-11a (NKP-15) positive cells. Only a small number of functional Leu-7 (HNK-1) antigen. In present study, we established techniques for immunoelectron microscopic identification by mouse monoclonal FITC-conjugated anti-Leu-11a biotinylated anti-Leu-7 antibodies. Ficoll-Hypaque-isolated peripheral blood lymphocytes (PBL) were reacted with specific antibodies before or after fixation in 1% glutaraldehyde/1% paraformaldehyde fixative. Prefixation labeling viable was carried out at 4°C 37°C. Cells prelabeled antibody secondary either fixation. Anti-Leu-7 stained directly via an avidin-biotin-peroxidase (ABC) system, indirectly ABC immunoperoxidase procedure anti-mouse IgG 10 nm 40 colloidal gold-labeled antibody. Results indicate that antigen could be localized incubation 20 min fixation; however, totally abrogated following same procedure. well methods prefixation system With respect gold labeling, better results obtained when immediately but Ultrastructurally both (+) shared common ultrastructural features associated large granular lymphocytes. Using above described techniques, found approximately 2–5% Leu-7+ 9–15% Leu-11a+ PBL healthy donors. overall suggest is more sensitive glutaraldehyde/paraformaldehyde than Leu-7, since it can only procedures; ideal technique light electron enumeration; immunogold method provides adequate which designated ultracythochemical studies.