Proliferative Response of Cloned CTL line 10B-5 upon Stimulation with Soluble Clonotypic Monoclonal Antibody and its Blocking by Anti-Lyt-2 Antibody

Heiichiro Udono, Kagemasa Kuribayashi, Teizo Fujita, Masahiro Mieno, Hiroshi Shiku, Eiichi Nakayama

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Proliferation of the cloned cytotoxic T lymphocyte (CTL) line 10B-5 induced by anti-clonotypic antibody and its blocking by anti-Lyt-2 mAb were studied. Clone 10B-5 was derived from spleen cells of a (BALB/c X CS7BL/6)F1 (CB6F1)-nu/+ mouse immunized with UV♀1 sarcoma. 10B-5 cells lysed UV♀1 specifically and proliferated upon stimulation with UV♀1 and feeder cells in the presence of IL2. 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♀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 IL2, 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.

Original languageEnglish
Pages (from-to)390-400
Number of pages11
JournalImmunobiology
Volume178
Issue number4-5
DOIs
Publication statusPublished - Jan 1 1989
Externally publishedYes

Keywords

  • (BALB/c X C57BL/6)F
  • B6
  • C57BL/6
  • CB6F
  • CTL
  • Con A
  • IL2
  • MHC
  • OVA
  • T cell antigen receptor
  • T cell growth factor
  • TCGF
  • TCR
  • UV
  • concanavalin A
  • cytotoxic T lymphocyte
  • interleukin 2
  • mAb
  • major histocompatibility complex
  • monoclonal antibody
  • ovalbumin
  • ultraviolet

ASJC Scopus subject areas

  • Immunology and Allergy
  • Immunology
  • Hematology

Fingerprint Dive into the research topics of 'Proliferative Response of Cloned CTL line 10B-5 upon Stimulation with Soluble Clonotypic Monoclonal Antibody and its Blocking by Anti-Lyt-2 Antibody'. Together they form a unique fingerprint.

  • Cite this