Open AccessAnalysis of CD8 T‐cell response by IFNγ ELISPOT and H‐2L d / pRL1a tetramer assays in pRL1a multiple antigen peptide‐immunized and RL male 1‐bearing BALB/ c and (BALB/c×C57BL/6) F 1 mice
Author(s) -
Takada Itsuro,
Noguchi Yuji,
Kenjo Akira,
Wada Hisashi,
Uenaka Akiko,
Fujita Teizo,
Inoue Hajime,
Nakayama Eiichi
Publication year - 2004
Publication title -
cancer science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.035
H-Index - 141
eISSN - 1349-7006
pISSN - 1347-9032
DOI - 10.1111/j.1349-7006.2004.tb02212.x
Subject(s) - elispot , cytotoxic t cell , cd8 , microbiology and biotechnology , antigen , t cell , spleen , biology , epitope , tetramer , chemistry , in vitro , immune system , immunology , biochemistry , enzyme
We previously identified an H‐2L d ‐binding peptide pRL1a (IPGLPLSL) on RL male 1 that is predominantly recognized by cytotoxic T‐lymphocytes (CTLs). MAP is a multibranched lysine core with antigenic peptides. Immunization of BALB/c mice with pRL1a MAP effectively induced pRL1a CTLs. Here, we demonstrate the presence of pRL1a‐recognizing CD8 + T‐cells in pRL1a MAP‐immunized and RL male 1‐bearing BALB/c and (BALB/ cxC57BL/6)F 1 mice by using IFNγ ELISPOT and H‐2L d /pRL1a tetramer assays. A few IFNγ ELISPOTs and no tetramer‐positive cells were detected ex vivo in spleen cells from BALB/c mice immunized with pRL1a MAP. After a single in vitro stimulation with RL male 1, 432 and 741 IFNγ ELISPOTs/10 5 cells were detected and tetramer‐positive CD8 + T‐cells occurred at relative frequencies of 5.7% and 30.8% in splenic CD8 + T‐cells from mice that had been doubly and triply immunized, respectively, against pRL1a MAP. Tetramer‐positive cells displayed two distinct cell populations, CD62L low and CD62L high . Secondary in vitro stimulation expanded CD62L high cells more efficiently than CD62L low cells. Furthermore, a higher frequency of IFNγ‐producing and tetramer‐positive CD8 + T‐cells was detected ex vivo in RL male 1‐bearing semi‐allogeneic (BALB/cxC57BL/6)F 1 than in BALB/c mice on day 14 after tumor inoculation.