Peptide‐pulsed dendritic cells induce tumoricidal cytotoxic T lymphocytes from healthy donors against stably HLA‐A*0201‐binding peptides from the Melan‐A/MART‐1 self antigen

The melanoma antigen Melan‐A/MART‐1 was screened for the presence of potential HLA‐A*0201‐binding cytotoxic T lymphocytes (CTL) epitopes. The immunodominant nonamer epitope AAGIGILTV demonstrated weak binding to T2 but a significant half‐life of binding to HLA‐A*0201 in contrast to the decamer EAAGIGILTV. In addition to the immunodominant CTL epitope, we describe two peptides, GILTVILGV and ALMDKSLHV, that display stable binding to HLA‐A*0201. Using cultured autologous dendritic cells pulsed with these peptides, CTL lines were induced from peripheral blood lymphocytes that displayed reactivity with HLA‐A2+, Melan‐A/MART‐1+ melanoma cells. CTL reactivity against the immunodominant epitope could be induced with the nonamer epitope alone, but not with the decamer variant. CTL clones generated from an (EAAGIGILTV + AAGIGILTV)‐induced CTL line recognize the appropriate melanoma cells and normal melanocytes. Upon further characterization of one of these CTL clones, it was found to be of surprisingly high affinity considering that it is directed against a self antigen. This study demonstrates that immunogenic peptides can be selected based on stability (half‐life) of peptide/HLA binding. In addition, cultured DC were found to efficiently induce CTL responses in vitro against such selected peptides, and some of these CTL were capable of recognizing endogenously processed antigen.

[1]  S. Rosenberg,et al.  Cloning of the gene coding for a shared human melanoma antigen recognized by autologous T cells infiltrating into tumor. , 1994, Proceedings of the National Academy of Sciences of the United States of America.

[2]  K. Sakaguchi,et al.  Identification of the immunodominant peptides of the MART-1 human melanoma antigen recognized by the majority of HLA-A2-restricted tumor infiltrating lymphocytes , 1994, The Journal of experimental medicine.

[3]  M. Serrano,et al.  A p16INK4a-insensitive CDK4 mutant targeted by cytolytic T lymphocytes in a human melanoma , 1995, Science.

[4]  E. Gilboa,et al.  Bone marrow-generated dendritic cells pulsed with a class I-restricted peptide are potent inducers of cytotoxic T lymphocytes , 1995, The Journal of experimental medicine.

[5]  S. H. van der Burg,et al.  Immunogenicity of peptides bound to MHC class I molecules depends on the MHC-peptide complex stability. , 1996, Journal of immunology.

[6]  M. Lotze,et al.  Mass spectrometric identification of a naturally processed melanoma peptide recognized by CD8+ cytotoxic T lymphocytes , 1995, The Journal of experimental medicine.

[7]  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.

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

[9]  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.

[10]  P. Coulie,et al.  A peptide encoded by human gene MAGE‐3 and presented by HLA‐A2 induces cytolytic T lymphocytes that recognize tumor cells expressing MAGE‐3 , 1994, European journal of immunology.

[11]  J. Shabanowitz,et al.  An HLA-A2-restricted tyrosinase antigen on melanoma cells results from posttranslational modification and suggests a novel pathway for processing of membrane proteins , 1996, The Journal of experimental medicine.

[12]  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.

[13]  L. Zitvogel,et al.  Bone marrow-derived dendritic cells pulsed with synthetic tumour peptides elicit protective and therapeutic antitumour immunity , 1995, Nature Medicine.

[14]  R. Steinman,et al.  Proliferating dendritic cell progenitors in human blood , 1994, The Journal of experimental medicine.

[15]  D. Margulies,et al.  Determinant selection of major histocompatibility complex class I- restricted antigenic peptides is explained by class I-peptide affinity and is strongly influenced by nondominant anchor residues , 1994, The Journal of experimental medicine.

[16]  M. Bednarek,et al.  The minimum peptide epitope from the influenza virus matrix protein. Extra and intracellular loading of HLA-A2. , 1991, Journal of immunology.

[17]  R. Steinman,et al.  Influenza virus-infected dendritic cells stimulate strong proliferative and cytolytic responses from human CD8+ T cells. , 1994, The Journal of clinical investigation.

[18]  P. Coulie,et al.  A mutated intron sequence codes for an antigenic peptide recognized by cytolytic T lymphocytes on a human melanoma. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[19]  T. Espevik,et al.  A highly sensitive cell line, WEHI 164 clone 13, for measuring cytotoxic factor/tumor necrosis factor from human monocytes. , 1986, Journal of immunological methods.

[20]  A. Vitiello,et al.  Definition of a minimal optimal cytotoxic T-cell epitope within the hepatitis B virus nucleocapsid protein , 1993, Journal of virology.

[21]  D. Kioussis,et al.  Low avidity recognition of self-antigen by T cells permits escape from central tolerance. , 1995, Immunity.

[22]  R. Versteeg,et al.  c‐myc down‐regulates class I HLA expression in human melanomas. , 1988, The EMBO journal.

[23]  P. Coulie,et al.  The tyrosinase gene codes for an antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas , 1993, The Journal of experimental medicine.

[24]  J. Renauld,et al.  A new gene coding for a differentiation antigen recognized by autologous cytolytic T lymphocytes on HLA-A2 melanomas , 1994, The Journal of experimental medicine.

[25]  A. Houghton,et al.  Implicating a role for immune recognition of self in tumor rejection: passive immunization against the brown locus protein , 1995, The Journal of experimental medicine.

[26]  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.

[27]  F. Sallusto,et al.  Efficient presentation of soluble antigen by cultured human dendritic cells is maintained by granulocyte/macrophage colony-stimulating factor plus interleukin 4 and downregulated by tumor necrosis factor alpha , 1994, The Journal of experimental medicine.

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

[29]  M. Feltkamp,et al.  Efficient MHC class I-peptide binding is required but does not ensure MHC class I-restricted immunogenicity. , 1994, Molecular immunology.

[30]  J. Zeuthen,et al.  A Human Melanoma Cell Line, Recognized by Both HLA Class I and Class II Restricted T Cells, is Capable of Initiating both Primary and Secondary Immune Responses , 1995, Scandinavian journal of immunology.

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

[32]  M. Ressing,et al.  CTL specific for the tyrosinase autoantigen can be induced from healthy donor blood to lyse melanoma cells. , 1995, Journal of immunology.

[33]  P. Bruggen,et al.  Tumor antigens recognized by T lymphocytes. , 1994, Annual review of immunology.