Peptide Fine Specificity of Anti-Glycoprotein 100 CTL Is Preserved Following Transfer of Engineered TCRαβ Genes Into Primary Human T Lymphocytes 1

TCR with known antitumor reactivity can be genetically introduced into primary human T lymphocytes and provide promising tools for immunogene therapy of tumors. We molecularly characterized two distinct TCRs specific for the same HLA-A2-restricted peptide derived from the melanocyte differentiation Ag gp100, yet exhibiting different stringencies in peptide requirements. The existence of these two distinct gp100-specific TCRs allowed us to study the preservation of peptide fine specificity of native TCRαβ when engineered for TCR gene transfer into human T lymphocytes. Retroviral transduction of primary human T lymphocytes with either one of the two sets of TCRαβ constructs enabled T lymphocytes to specifically kill and produce TNF-α when triggered by native gp100pos/HLA-A2pos tumor target cells as well as gp100 peptide-loaded HLA-A2pos tumor cells. Peptide titration studies revealed that the cytolytic efficiencies of the T lymphocyte transductants were in the same range as those of the parental CTL clones. Moreover, primary human T lymphocytes expressing either one of the two engineered gp100-specific TCRs show cytolytic activities in response to a large panel of peptide mutants that are identical with those of the parental CTL. The finding that two gp100-specific TCR, derived from two different CTL, can be functionally introduced into primary human T lymphocytes without loss of the Ag reactivity and peptide fine specificity, holds great promise for the application of TCR gene transfer in cancer immunotherapy.

[1]  R. Henderson,et al.  Identification of a peptide recognized by five melanoma-specific human cytotoxic T cell lines. , 1994, Science.

[2]  Bolhuis Rl,et al.  Genetic re-targeting of T lymphocyte specificity. , 1998 .

[3]  E. Kawasaki,et al.  A general method for facilitating heterodimeric pairing between two proteins: application to expression of alpha and beta T-cell receptor extracellular segments. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[4]  G. Rovera,et al.  Molecular diagnosis of cutaneous T-cell lymphoma: polymerase chain reaction amplification of T-cell antigen receptor beta-chain gene rearrangements. , 1991, The Journal of investigative dermatology.

[5]  Steven A. Rosenberg,et al.  Progress in human tumour immunology and immunotherapy , 2001, Nature.

[6]  M. Theobald,et al.  Tolerance to p53 by A2.1-restricted Cytotoxic T Lymphocytes , 1997, The Journal of experimental medicine.

[7]  Partho Ghosh,et al.  Structure of the complex between human T-cell receptor, viral peptide and HLA-A2 , 1996, Nature.

[8]  S. H. van der Burg,et al.  Identification of peptide sequences that potentially trigger HLA‐A2.1‐restricted cytotoxic T lymphocytes , 1993, European journal of immunology.

[9]  Robert H. Collins,et al.  Donor leukocyte infusions in 140 patients with relapsed malignancy after allogeneic bone marrow transplantation. , 1997, Journal of clinical oncology : official journal of the American Society of Clinical Oncology.

[10]  Tak W. Mak,et al.  Human T-cell receptor variable gene segment families , 1995, Immunogenetics.

[11]  M. Kalos,et al.  Transfer of Specificity for Human Immunodeficiency Virus Type 1 into Primary Human T Lymphocytes by Introduction of T-Cell Receptor Genes , 2000, Journal of Virology.

[12]  M. Matsuda,et al.  Decreased expression of the signal-transducing zeta chains in tumor-infiltrating T-cells and NK cells of patients with colorectal carcinoma. , 1993, Cancer research.

[13]  A. Lesk,et al.  The outline structure of the T‐cell alpha beta receptor. , 1988, The EMBO journal.

[14]  D. Longo,et al.  Loss of T-cell receptor zeta chain and p56lck in T-cells infiltrating human renal cell carcinoma. , 1993, Cancer research.

[15]  S. H. van der Burg,et al.  Affinity, specificity and T‐cell‐receptor diversity of melanoma‐specific CTL generated in vitro against a single tyrosinase epitope , 1997, International journal of cancer.

[16]  P. Henkart,et al.  Cytotoxic cells : basic mechanisms and medical applications , 2000 .

[17]  J. Sidney,et al.  Prominent role of secondary anchor residues in peptide binding to HLA-A2.1 molecules , 1993, Cell.

[18]  T. Schumacher,et al.  Immunotherapy through TCR gene transfer , 2001, Nature Immunology.

[19]  S. H. van der Burg,et al.  An HLA class I peptide-binding assay based on competition for binding to class I molecules on intact human B cells. Identification of conserved HIV-1 polymerase peptides binding to HLA-A*0301. , 1995, Human immunology.

[20]  S. Rosenberg,et al.  Efficient transfer of a tumor antigen-reactive TCR to human peripheral blood lymphocytes confers anti-tumor reactivity. , 1999, Journal of immunology.

[21]  C. Lamers,et al.  Activation of the immune system of cancer patients by continuous i.v. recombinant IL‐2 (rIL‐2) therapy is dependent on dose and schedule of rIL‐2 , 1993, Clinical and experimental immunology.

[22]  R. Schreiber,et al.  Eradication of established tumors by CD8+ T cell adoptive immunotherapy. , 2000, Immunity.

[23]  Z. Eshhar,et al.  Harnessing Syk family tyrosine kinases as signaling domains for chimeric single chain of the variable domain receptors: optimal design for T cell activation. , 1998, Journal of immunology.

[24]  S. Rosenberg,et al.  Shared human melanoma antigens. Recognition by tumor-infiltrating lymphocytes in HLA-A2.1-transfected melanomas. , 1992, Journal of immunology.

[25]  H. Zarour,et al.  The majority of autologous cytolytic T-lymphocyte clones derived from peripheral blood lymphocytes of a melanoma patient recognize an antigenic peptide derived from gene Pmel17/gp100. , 1996, The Journal of investigative dermatology.

[26]  H. Eisen,et al.  Evidence that a single peptide-MHC complex on a target cell can elicit a cytolytic T cell response. , 1996, Immunity.

[27]  J. Sidney,et al.  Identification of subdominant CTL epitopes of the GP100 melanoma-associated tumor antigen by primary in vitro immunization with peptide-pulsed dendritic cells. , 1997, Journal of immunology.

[28]  J W Gratama,et al.  Grafting primary human T lymphocytes with cancer-specific chimeric single chain and two chain TCR , 2000, Gene Therapy.

[29]  S. Rosenberg,et al.  Characterization of the functional specificity of a cloned T-cell receptor heterodimer recognizing the MART-1 melanoma antigen. , 1995, Cancer research.

[30]  M. Weijtens,et al.  A retroviral vector system ‘STITCH’ in combination with an optimized single chain antibody chimeric receptor gene structure allows efficient gene transduction and expression in human T lymphocytes , 1998, Gene Therapy.

[31]  R. Steinman,et al.  The distinct surface of human blood dendritic cells, as observed after an improved isolation method. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[32]  Hans J. Stauss,et al.  Circumventing tolerance to a human MDM2-derived tumor antigen by TCR gene transfer , 2001, Nature Immunology.

[33]  T. Kitamura,et al.  Functional Reconstitution of Class II MHC-Restricted T Cell Immunity Mediated by Retroviral Transfer of the αβ TCR Complex1 , 2000, The Journal of Immunology.

[34]  S. H. van der Burg,et al.  Analogues of CTL epitopes with improved MHC class‐I binding capacity elicit anti‐melanoma CTL recognizing the wild‐type epitope , 1997, International journal of cancer.

[35]  P. Romero,et al.  Multiple specificities in the repertoire of a melanoma patient's cytolytic T lymphocytes directed against tumor antigen MAGE-1.A1 , 1995, The Journal of experimental medicine.

[36]  C. Figdor,et al.  Melanocyte lineage-specific antigen gp100 is recognized by melanoma- derived tumor-infiltrating lymphocytes , 1994, The Journal of experimental medicine.

[37]  R. Steinman,et al.  Antigen-specific T lymphocytes efficiently cluster with dendritic cells in the human primary mixed-leukocyte reaction. , 1988, Cellular immunology.

[38]  D. Diamond,et al.  Targeting of human p53-overexpressing tumor cells by an HLA A*0201-restricted murine T-cell receptor expressed in Jurkat T cells. , 2000, Cancer research.

[39]  David A. Williams,et al.  High-Efficiency Gene Transfer into Normal and Adenosine Deaminase-Deficient T Lymphocytes Is Mediated by Transduction on Recombinant Fibronectin Fragments , 1998, Journal of Virology.

[40]  B. V. van Krimpen,et al.  Rapid expansion of human cytotoxic T cell clones: growth promotion by a heat-labile serum component and by various types of feeder cells. , 1984, Journal of immunological methods.

[41]  J. McCubrey,et al.  Transfer of specificity by murine α and β T-cell receptor genes , 1986, Nature.

[42]  S. Rosenberg.  A new era of cancer immunotherapy: Converting theory to performance , 1999, CA: a cancer journal for clinicians.

[43]  J. Bell,et al.  Comparative sequence analysis of the human T cell receptor TCRA and TCRB CDR3 regions. , 1996, Human immunology.

[44]  H. Heslop,et al.  Adoptive cellular immunotherapy for EBV lymphoproliferative diseases , 1997 .

[45]  K. Sakaguchi,et al.  Identification of a human melanoma antigen recognized by tumor-infiltrating lymphocytes associated with in vivo tumor rejection. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[46]  H. Heslop,et al.  Adoptive cellular immunotherapy for EBV lymphoproliferative disease. , 1997, Immunological reviews.

[47]  G. Nolan,et al.  Retroviral transduction of a T cell receptor specific for an Epstein-Barr virus-encoded peptide. , 2001, Clinical immunology.

[48]  C. Figdor,et al.  Generation of antimelanoma cytotoxic T lymphocytes from healthy donors after presentation of melanoma-associated antigen-derived epitopes by dendritic cells in vitro. , 1995, Cancer research.

[49]  P. Linsley,et al.  Regulation of immunostimulatory function and costimulatory molecule (B7-1 and B7-2) expression on murine dendritic cells. , 1994, Journal of immunology.

[50]  P. Robbins,et al.  A listing of human tumor antigens recognized by T cells , 2001, Cancer Immunology, Immunotherapy.