title: The cytotoxic T cell response to peptide analogs of the HLA-A*0201-restricted MUC1 signal sequence epitope, M1.2.
abstract: The mucin MUC1 molecule is overexpressed on a variety of adenocarcinomas and is thus, a potential target for immunotherapy. Of the MUC1 peptides that bind to HLA-A*0201(A2), M1.2 (LLLLTVLTV) from the signal sequence appears to be the most immunogenic in humans. Here we have shown that large numbers (10(9)) of tetramer-binding M1.2-specific cytotoxic T lymphocytes (CTL) can be generated ex vivo from circulating precursors, derived from healthy adults. However, there was significant interpersonal variation in the level of co-stimulatory signal required. Tetramer-binding cells also required maturation in culture to become proficient killers of the HLA-A2(+) MUC1(+) MCF7 cell line, known to express a low number of endogenously processed M1.2. The functional avidity of M1.2-specific CTL, however, was low as compared to CTL specific for an HIV-1 epitope. Despite the low avidity, M1.2-specific CTL were polyfunctional, secreting multiple cytokines upon degranulation with antigen recognition. To identify potential agonist peptides that may be superior immunogens, an M1.2-specific CTL culture was used to scan a large nonameric combinatorial peptide library. Of 54 predicted peptides, 4 were "consensus" agonists because they were recognized by CTL from two other donors. Two agonists, p29 (LLPWTVLTV) and p15 (VLLWTVLTV), were equally stimulatory when loaded onto C1R target cells transfected with wild-type HLA-A2. Both agonists induced IL-2, TNF-alpha, IFN-gamma, and degranulation with M1.2-specific CTL. In contrast, production of these cytokines, which are tightly regulated by specific activation through the T cell receptor, was restricted when the CTL were stimulated with peptides loaded onto C1R cells that were transfected with an HLA-A2 molecule bearing a mutation that abrogates binding to the CD8 co-receptor. Thus, activation by both M1.2 and its agonists was dependent upon CD8, showing that compensation by the co-receptor was necessary for the human T cell response to M1.2.
authors.affiliation: Department of Medicine, Wayne State University School of Medicine, Detroit, MI, 48201, USA.
authors.forename: Malcolm S
authors.initials: MS
authors.lastname: Mitchell
authors.forename: Teri A
authors.initials: TA
authors.lastname: Lund
authors.forename: Andrew K
authors.initials: AK
authors.lastname: Sewell
authors.forename: Francesco M
authors.initials: FM
authors.lastname: Marincola
authors.forename: Elyse
authors.initials: E
authors.lastname: Paul
authors.forename: Kim
authors.initials: K
authors.lastname: Schroder
authors.forename: Darcy B
authors.initials: DB
authors.lastname: Wilson
authors.forename: June
authors.initials: J
authors.lastname: Kan-Mitchell
mesh_terms: D000328:Adult; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D003713:Dendritic Cells / Q000187:drug effects; D018984:Epitopes, T-Lymphocyte / Q000187:drug effects / Q000276:immunology*; D005260:Female; D015234:HLA-A Antigens / Q000276:immunology*; D015789:HLA-A2 Antigen; D006801:Humans; D008070:Lipopolysaccharides / Q000494:pharmacology; D008297:Male; D009000:Monocytes / Q000187:drug effects; D018396:Mucin-1 / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology* / Q000494:pharmacology; D019151:Peptide Library; D012016:Reference Values; D015398:Signal Transduction / Q000187:drug effects / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000187:drug effects / Q000276:immunology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
chemical_list: D018984:Epitopes, T-Lymphocyte; D015234:HLA-A Antigens; C435939:HLA-A*02:01 antigen; D015789:HLA-A2 Antigen; D008070:Lipopolysaccharides; D018396:Mucin-1; D010446:Peptide Fragments; D019151:Peptide Library; D014409:Tumor Necrosis Factor-alpha