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Proliferative response of cloned CTL line 10B-5 upon stimulation with soluble clonotypic monoclonal antibody and its blocking by anti-Lyt-2 antibody.
- Source :
-
Immunobiology [Immunobiology] 1989 Feb; Vol. 178 (4-5), pp. 390-400. - Publication Year :
- 1989
-
Abstract
- 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.
- Subjects :
- Animals
Clone Cells
Female
Immunoglobulin Fab Fragments immunology
Interleukin-2 pharmacology
Male
Mice
Mice, Inbred BALB C
Mice, Inbred C57BL
Receptors, Antigen, T-Cell immunology
Recombinant Proteins pharmacology
Antibodies, Monoclonal immunology
Isoantibodies immunology
Lymphocyte Activation
T-Lymphocytes, Cytotoxic immunology
Subjects
Details
- Language :
- English
- ISSN :
- 0171-2985
- Volume :
- 178
- Issue :
- 4-5
- Database :
- MEDLINE
- Journal :
- Immunobiology
- Publication Type :
- Academic Journal
- Accession number :
- 2785484
- Full Text :
- https://doi.org/10.1016/S0171-2985(89)80061-0