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Recognition of breast cancer‐associated peptides by tumor‐reactive, HLA‐class I restricted allogeneic cytotoxic T lymphocytes
Author(s) -
Nguyen Thelinh,
Naziruddin Bashoo,
Dintzis Suzanne,
Doherty Gerard M.,
Mohanakumar T.
Publication year - 1999
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/(sici)1097-0215(19990517)81:4<607::aid-ijc17>3.0.co;2-x
Subject(s) - ctl* , cytotoxic t cell , cd8 , tumor infiltrating lymphocytes , immunology , biology , human leukocyte antigen , breast cancer , cancer research , monoclonal antibody , antigen , cancer , antibody , in vitro , biochemistry , genetics
Strategies to identify tumor‐associated antigens rely on the paradigm that tumor‐associated peptides presented in the context of HLA‐class I are recognized by the cellular immune system. Approaches to isolate tumor‐specific cytotoxic T lymphocytes (CTL) from tumor‐infiltrating lymphocytes are difficult because long‐term growth of the CTL requires autologous tumor cells and lymphocytes (PBL) as feeder cells. In this study, a CTL line (BL.HBL‐100 CTL) was generated from PBL from a normal healthy donor by stimulating with irradiated, HLA‐class I partially matched breast cancer cell line HBL‐100. Activated T lymphocytes generated expressed TCRα/β + with a predominant CD8 + population after 12 stimulations (98.54% CD8 + vs. 0.18% CD4 + ). These CTL lysed HLA‐A1 + , but not HLA‐A1 − , breast cancer cell lines. Moreover, HLA‐A1 + , non breast cancer cell lines were not recognized. The lytic activity of BL.HBL‐100 CTL against HLA‐A1 + breast cancer cell lines was blocked by monoclonal antibodies (MAbs) to HLA‐class I and CD8, but not by anti‐HLA‐class II and CD4. Recognition of HLA‐A1 + breast cancer cells by the CTL was dependent on peptides associated with HLA‐class I since the lysis was inhibited by acid elution of HLA bound peptides. HBL‐100 tumors were grown in severe combined immunodeficient (SCID) mice. Immunohistochemical staining of the HBL‐100 tissue harvested from SCID mice demonstrated human breast cancer cells. HLA‐class I molecules were affinity purified from the HBL‐100 harvested from the SCID mice; class I bound peptides were eluted and separated by RP‐HPLC. Pooled HPLC peptide fractions were tested for reconstituting antigenic epitopes recognized by the BL.HBL‐100 CTL and found to reside within fraction 40. Our results show that a tumor reactive, HLA‐class I restricted CTL was produced by stimulating normal PBL against an HLA‐class I matched breast cancer cell line. We also provide evidence for a breast cancer‐associated, HLA‐class I bound peptide antigen(s) that reconstitutes the antigenic epitope(s) recognized by these CTL. Int. J. Cancer 81:607–615, 1999. © 1999 Wiley‐Liss, Inc.