Proliferative response of cloned CTL line 10B-5 upon stimulation with soluble clonotypic monoclonal antibody and its blocking by anti-Lyt-2 antibody.
Proliferation of the cloned cytotoxic T lymphocyte (CTL) line 10B-5 induced by anticlonotypic antibody and its blocking by anti-Lyt-2 mAb were studied. Clone 10B-5 was derived from spleen cells of a (BALB/c x C57BL/6)F1, (CB6F1)-nu/+ mouse immunized with UV female 1 sarcoma. 10B-5 cells lysed UV female 1 specifically and proliferated upon stimulation with UV female 1 and feeder cells in the presence of IL 2. Anti-clonotypic monoclonal antibody (mAb) N1-56 was produced by a hybridoma established by fusion of spleen cells from a CB6F1-nu/+ mouse that had been immunized by five injections of 10B-5 cells prefixed with 0.1% formaldehyde. N1-56 mAb immunoprecipitated 90 kd molecules cleavable to 45 kd molecules under reducing conditions, indicating its reaction with T cell antigen receptor (TCR). N1-56 mAb in its soluble form induced a proliferative response of 10B-5 cells. Thus, the antigen binding site of N1-56 mAb appeared to substitute for the specific antigen determinant on UV female 1 sarcoma. The F(ab')2, but not the Fab fragment of purified N1-56 mAb, stimulated proliferation, indicating that cross-linking of TCR molecules was necessary for stimulation. The proliferative response of 10B-5 cells induced by soluble N1-56 mAb was blocked by addition of anti-Lyt-2.2 mAb to the cultures. The specificity of Lyt-2 blocking was confirmed by an absorption test. The proliferative response of 10B-5 cells induced by Con A, but not that induced by IL 2, was blocked by anti-Lyt-2.2 mAb. These results indicated that blocking by anti-Lyt-2.2 mAb was at an early stage and that it could be bypassed by stimulation with IL2.
Udono
H
H
Department of Oncology, Nagasaki University School of Medicine, Japan.
Kuribayashi
K
K
Fujita
T
T
Mieno
M
M
Shiku
H
H
Nakayama
E
E
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D002999:Clone Cells; D005260:Female; D007140:Immunoglobulin Fab Fragments / Q000276:immunology; D007376:Interleukin-2 / Q000494:pharmacology; D007518:Isoantibodies / Q000276:immunology*; D008213:Lymphocyte Activation; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D008810:Mice, Inbred C57BL; D011948:Receptors, Antigen, T-Cell / Q000276:immunology; D011994:Recombinant Proteins / Q000494:pharmacology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*
D000911:Antibodies, Monoclonal; D007140:Immunoglobulin Fab Fragments; D007376:Interleukin-2; D007518:Isoantibodies; C029805:Lyt antibodies; D011948:Receptors, Antigen, T-Cell; D011994:Recombinant Proteins