Vaccination of Cancer Patients Against Telomerase Induces Functional Antitumor CD8+ T Lymphocytes

Purpose: High-level expression of the telomerase reverse transcriptase (hTERT) in >85% of human cancers, in contrast with its restricted expression in normal adult tissues, points to hTERT as a broadly applicable molecular target for anticancer immunotherapy. CTLs recognize peptides derived from hTERT and kill hTERT+ tumor cells of multiple histologies in vitro. Moreover, because survival of hTERT+ tumor cells requires functionally active telomerase, hTERT mutation or loss as a means of escape may be incompatible with sustained tumor growth. Experimental Design: A Phase I clinical trial was performed to evaluate the clinical and immunological impact of vaccinating advanced cancer patients with the HLA-A2-restricted hTERT I540 peptide presented with keyhole limpet hemocyanin by ex vivo generated autologous dendritic cells. Results: As measured by peptide/MHC tetramer, enzyme-linked immunospot, and cytotoxicity assays, hTERT-specific T lymphocytes were induced in 4 of 7 patients with advanced breast or prostate carcinoma after vaccination with dendritic cells pulsed with hTERT peptide. Tetramer-guided high-speed sorting and polyclonal expansion achieved highly enriched populations of hTERT-specific cells that killed tumor cells in an MHC- restricted fashion. Despite concerns of telomerase activity in rare normal cells, no significant toxicity was observed. Partial tumor regression in 1 patient was associated with the induction of CD8+ tumor infiltrating lymphocytes. Conclusions: These results demonstrate the immunological feasibility of vaccinating patients against telomerase and provide rationale for targeting self-antigens with critical roles in oncogenesis.

[1]  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.

[2]  R. Vonderheide.  Telomerase as a universal tumor-associated antigen for cancer immunotherapy , 2002, Oncogene.

[3]  R. Glasspool,et al.  Telomerase-specific suicide gene therapy vectors expressing bacterial nitroreductase sensitize human cancer cells to the pro-drug CB1954 , 2001, Oncogene.

[4]  B. Walker,et al.  An optimal viral peptide recognized by CD8+ T cells binds very tightly to the restricting class I major histocompatibility complex protein on intact cells but not to the purified class I protein. , 1991, Proceedings of the National Academy of Sciences of the United States of America.

[5]  K. Skorecki,et al.  Isolation and characterization of human recombinant antibodies endowed with the antigen-specific, major histocompatibility complex-restricted specificity of T cells directed toward the widely expressed tumor T-cell epitopes of the telomerase catalytic subunit. , 2002, Cancer research.

[6]  Philip J. R. Goulder,et al.  Phenotypic Analysis of Antigen-Specific T Lymphocytes , 1996, Science.

[7]  Francesco M Marincola,et al.  Cancer immunotherapy with peptide-based vaccines: what have we achieved? Where are we going? , 2002, Journal of the National Cancer Institute.

[8]  M. Rep,et al.  Phenotypic and Functional Separation of Memory and Effector Human CD8+ T Cells , 1997, The Journal of experimental medicine.

[9]  J. Shay,et al.  Human Telomerase and Its Regulation , 2002, Microbiology and Molecular Biology Reviews.

[10]  W. Hahn,et al.  Inhibition of telomerase limits the growth of human cancer cells , 1999, Nature Medicine.

[11]  David Allman,et al.  Ex vivo expansion of polyclonal and antigen-specific cytotoxic T lymphocytes by artificial APCs expressing ligands for the T-cell receptor, CD28 and 4-1BB , 2002, Nature Biotechnology.

[12]  M. Robinson,et al.  Telomere shortening and apoptosis in telomerase-inhibited human tumor cells. , 1999, Genes & development.

[13]  R. Steinman,et al.  Rapid generation of broad T-cell immunity in humans after a single injection of mature dendritic cells. , 1999, The Journal of clinical investigation.

[14]  D. Schadendorf,et al.  Intranodal injection of semimature monocyte-derived dendritic cells induces T helper type 1 responses to protein neoantigen. , 2003, Blood.

[15]  A. Hasegawa,et al.  Identification of human telomerase reverse transcriptase-derived peptides that induce HLA-A24-restricted antileukemia cytotoxic T lymphocytes. , 2001, Blood.

[16]  P. Klenerman,et al.  Immune surveillance against a solid tumor fails because of immunological ignorance. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[17]  Eli Gilboa,et al.  Induction of cytotoxic T cell responses and tumor immunity against unrelated tumors using telomerase reverse transcriptase RNA transfected dendritic cells. , 2000, Nature Medicine.

[18]  Lieping Chen,et al.  Immunological ignorance of silent antigens as an explanation of tumor evasion. , 1998, Immunology today.

[19]  D. Schadendorf,et al.  Circulating Tumor-reactive CD8(+) T cells in melanoma patients contain a CD45RA(+)CCR7(-) effector subset exerting ex vivo tumor-specific cytolytic activity. , 2002, Cancer research.

[20]  W. Oyen,et al.  Effective migration of antigen-pulsed dendritic cells to lymph nodes in melanoma patients is determined by their maturation state. , 2003, Cancer research.

[21]  A. Enk,et al.  Vaccination with Mage-3a1 Peptide–Pulsed Mature, Monocyte-Derived Dendritic Cells Expands Specific Cytotoxic T Cells and Induces Regression of Some Metastases in Advanced Stage IV Melanoma , 1999, The Journal of experimental medicine.

[22]  F. Sallusto,et al.  Two subsets of memory T lymphocytes with distinct homing potentials and effector functions , 1999, Nature.

[23]  J. Shay,et al.  Telomerase: a target for cancer therapeutics. , 2002, Cancer cell.

[24]  R. Steinman,et al.  Immune and clinical responses in patients with metastatic melanoma to CD34(+) progenitor-derived dendritic cell vaccine. , 2001, Cancer research.

[25]  W. Hahn,et al.  Equivalent induction of telomerase-specific cytotoxic T lymphocytes from tumor-bearing patients and healthy individuals. , 2001, Cancer research.

[26]  M. Raffeld,et al.  Cancer Regression and Autoimmunity in Patients After Clonal Repopulation with Antitumor Lymphocytes , 2002, Science.

[27]  J. Hipp,et al.  Cytotoxic T cell immunity against telomerase reverse transcriptase in humans. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[28]  Javier Hernández,et al.  Identification of a human telomerase reverse transcriptase peptide of low affinity for HLA A2.1 that induces cytotoxic T lymphocytes and mediates lysis of tumor cells , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[29]  A. Enk,et al.  A comparison of two types of dendritic cell as adjuvants for the induction of melanoma‐specific T‐cell responses in humans following intranodal injection , 2001, International journal of cancer.

[30]  D. Corey,et al.  Inhibition of human telomerase in immortal human cells leads to progressive telomere shortening and cell death. , 1999, Proceedings of the National Academy of Sciences of the United States of America.

[31]  Jingwu Z. Zhang,et al.  Identification of HLA DR7-restricted epitopes from human telomerase reverse transcriptase recognized by CD4+ T-helper cells. , 2002, Cancer research.

[32]  D. Birx,et al.  Adoptive transfer of costimulated CD4+ T cells induces expansion of peripheral T cells and decreased CCR5 expression in HIV infection , 2002, Nature Medicine.

[33]  D. Corey.  Telomerase inhibition, oligonucleotides, and clinical trials , 2002, Oncogene.

[34]  Dirk Schadendorf,et al.  Vaccination of melanoma patients with peptide- or tumorlysate-pulsed dendritic cells , 1998, Nature Medicine.

[35]  C B Harley,et al.  Specific association of human telomerase activity with immortal cells and cancer. , 1994, Science.

[36]  Nina Bhardwaj,et al.  Antigen-Specific Inhibition of Effector T Cell Function in Humans after Injection of Immature Dendritic Cells , 2001, The Journal of experimental medicine.

[37]  J. Schultze,et al.  From cancer genomics to cancer immunotherapy: toward second-generation tumor antigens. , 2001, Trends in immunology.

[38]  W. Hahn,et al.  Telomerase activation, cellular immortalization and cancer , 2001, Annals of medicine.

[39]  D. Speiser,et al.  Lack of tumor recognition by hTERT peptide 540–548‐specific CD8+ T cells from melanoma patients reveals inefficient antigen processing , 2001, European journal of immunology.

[40]  R. Steinman,et al.  Active immunization against cancer with dendritic cells: The near future , 2001, International journal of cancer.

[41]  W. Hahn,et al.  Characterization of HLA-A3-restricted cytotoxic T lymphocytes reactive against the widely expressed tumor antigen telomerase. , 2001, Clinical cancer research : an official journal of the American Association for Cancer Research.

[42]  C. Greider,et al.  Developmental and tissue-specific regulation of mouse telomerase and telomere length. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[43]  W. Hahn,et al.  The telomerase catalytic subunit is a widely expressed tumor-associated antigen recognized by cytotoxic T lymphocytes. , 1999, Immunity.

[44]  H. Inoko,et al.  Target epitope in the Tax protein of human T-cell leukemia virus type I recognized by class I major histocompatibility complex-restricted cytotoxic T cells , 1992, Journal of virology.

[45]  F. Oesch,et al.  Inverse relationship of melanocyte differentiation antigen expression in melanoma tissues and CD8+ cytotoxic‐T‐cell responses: Evidence for immunoselection of antigen‐loss variants in vivo , 1996, International journal of cancer.