Prof. Daniel E. Speiser & Dr. Nathalie Rufer - Analysis of in vivo differentiation and function, and molecular dissection of antigen specific CD8 T cells before and after immunotherapy of melanoma patients

Dr. Nathalie Rufer

Head of Research Unit

Department of Research (DFR), CHUV, University of Lausanne, Switzerland
Ludwig Institute for Cancer Research, Lausanne, Switzerland

Dr. Nathalie Rufer received her PhD from the University of Geneva in 1996. She carried out a post-doctoral work in the laboratory of Peter Lansdorp (Terry Fox Laboratory, Vancouver, Canada) on telomere length biology in human T lymphocytes. In 2001, she joined ISREC as an associate scientist in the NCCR Molecular Oncology program and since 2009, she has been appointed by the CHUV-University of Lausanne. Her research interests are focused on human T lymphocyte responses against tumor antigens in melanoma patients, following therapeutic vaccination as well as on the understanding of the structural basis of T cell receptor affinity/avidity for antigen

Prof. Daniel E. Speiser

Member of the Ludwig Institute for Cancer Research

Prof. Daniel Speiser graduated in 1982 and received a Doctorate in Medicine in 1986 at the University of Zürich, Switzerland. Besides education in internal medicine, he trained for 5 years in the laboratory of R.M. Zinkernagel (Medical Faculty of Zürich), specializing in infectious and tumor immunity. He extended his experience to transplantation immunology and habilitated at the University of Geneva in 1995. Afterwards he developed experimental immunotherapy in a mouse model with naturally arising pancreatic tumors in the laboratories of P.S. Ohashi and T.W. Mak (University of Toronto). Since 1997 he is a Member of the Ludwig Institute for Cancer Research, Lausanne branch, where he leads the Clinical Immunotherapy Trial Program in Oncology at the University Hospital Lausanne. His research is focused on tumor antigens specific for melanoma and carcinomas, and activation, differentiation and function of T cells. The clinical studies have the aim to identify the most promising vaccine components promoting those human T cell responses which are strong and adequate to fight human cancer.

Département de formation et de recherche, CHUV

Ludwig Institute for Cancer Research

University of Lausanne

C/o HO Niv 5, Aile Est, 1532

Avenue Pierre-Decker, 4

1011 Lausanne

Switzerland

Tel Dr. Rufer: +41/21 314 01 99

Tel Prof. Speiser: +41/21 314 01 82

Fax: +41/21 314 74 77

Email:

Molecular analysis of T cell immune responses in melanoma patients

T lymphocytes and skin cancer

T cells are white blood cells representing an essential part of the immune system. They recognize and destroy any diseased cell which has been altered, e.g. by infection with a virus or by transformation into cancer cells. Melanoma has proven to be one of the most immunogenic among human tumors. A relatively high number of molecularly defined antigens that are recognized by T lymphocytes have been identified in the last 15 years. For instance, the differentiation antigen Melan-A is selectively expressed by normal cells of the melanocytic lineage including skin melanocytes and by a large majority of malignant melanomas. In the human melanoma cancer model, tumor antigen-specific T cell responses often develop, but their protective effects fail to control or eliminate the disease. This emphasizes the need to enhance our understanding of issues such as the identification of the type of T cells that are most efficient in anti-cancer therapy, and their functional properties in well-defined clinical situations.

T lymphocytes bear antigen-specific T cell receptors

Each T cell possesses a unique entity called T cell receptor. The main function of this receptor is to recognize target molecules of viral- or tumor-origin that are presented at the surface of a target cell by self-MHC class I molecules. A relatively small number of T cell precursor cells is normally selected in response to viral antigens. While well-defined antiviral-specific T cell responses consist of T cells expressing T cell receptors endowed with high avidity for their antigen ligand, T cells directed against tumor antigens express T cell receptors of lower avidity. The relative lack of high avidity T cell receptors may be a major reason why immune responses towards tumor antigens are often non-protective and T cell activity against tumor progression remains inefficient.

Cancer immunotherapy

Therapeutic vaccination has the aim of generating and/or boosting immune responses to destroy tumor cells and prevent tumor progression. Activated T cells proliferate and differentiate into “effector” cells, which secrete cytokines and exert cytolytic activity. Ongoing research aims to determine the specific roles of each subpopulation, and to identify vaccine components that support those T cells, which are central for efficient immune protection. Moreover, immunotherapy provides a unique opportunity to study T cell priming and memory formation in humans. Three aspects of the dynamics of T cell mediated immunity are of prime interest to us; (i) the identification of the type of T cells and their T cell receptors that are most efficient in anti-cancer therapy, (ii) the T cell receptor dynamics in patients with melanoma, and (iii) the structural basis of T cell receptor avidity for tumor antigens. The main goal of our projects is to improve the possibilities for using tumor-specific cytolytic T cells in anti-cancer therapy.

Tracking tumor-specific T lymphocytes in patients

We have developed new-sophisticated techniques that allow us to carefully dissect and follow human immune responses against tumors. For the first time, we are able to isolate anti-tumor T cells bearing tumor-specific T cell receptors with high potency for tumor cell recognition and destruction. Using this novel molecular-based strategy, we have recently tracked and followed dominant tumor-specific T cell clones in 15 melanoma patients over extended period of time. The identification of those “super” anti-tumor selected T cell receptors and their detailed characterization at the single cell level are key steps to better understand the relation between structural and functional features of T cell receptors to efficiently recognize tumor cells. Our approach of T cell analysis is much more extensive than what is usually done in humans or even in animal models. Expected results should allow streamlining the development of antigen specific immunotherapy against cancer and/or infections.

Figure 1. Human tumor-specific cytolytic T lymphocytes can be differentiated into functionally different sub-populations using a combination of cell surface markers such as tyrosinase phosphatase CD45RA and homing chemokine CCR7. Tumor-specific T lymphocytes bear a naive phenotype when analyzed at an early time-point before the start of peptide vaccination (CD45RA+CCR7+). In contrast, after vaccination, these T-cells undergo phenotypic and functionally changes in vivo, and become primed cells (CD45RA-CCR7-).

Figure 2. Telomere length analysis of hTERT-transduced T lymphocytes by in situ hybridization of metaphase chromosomes. We used a Peptide Nucleic Acid (PNA) probe that is complementary to the telomeric TTAGGG sequence. Cy-3-labeled telomeres are shown in yellow, while DAPI-stained chromosomes are depicted in blue. Telomere length is proportional to the fluorescent signal. Chromosomes with short telomeres were observed in T cell lymphocytes transduced with a control vector (A, arrows). In contrast, ectopic expression of hTERT had a direct effect on each chromosome by preventing the accumulation of short telomeres and significantly extending the replicative life span of T cells (B).

List of publications Dr. Rufer

Published papers with peer reviews

Wieckowski S, Baumgaertner P, Corthesy P, Voelter V, Romero P, Speiser DE, and Rufer N. Fine structural variations of abTCRs selected by vaccination with natural versus altered self-antigen in melanoma patients. J Immunol. 183(8):5397-406. (2009)

Jandus C, Bioley G, Dojcinovic D, Derré L, Baitsch L, Wieckowski S, Rufer N, Kwok WW, Tiercy J-M, Luescher IF, Speiser DE and Romero P. Tumor antigen specific regulatory CD4 T cells identified in human metastatic melanoma: peptide vaccination results in selective expansion of Th1-like counterparts. Cancer Res. (2009)

Iancu EM, Corthesy P, Baumgaertner P, Devevre E, Voelter V, Romero P, Speiser DE, and Rufer N. Clonotype selection and composition of human CD8 T cells specific for persistent herpes viruses varies with differentiation but is stable over time. J Immunol. 183(1):319-31 (2009)

Touvrey C, Derré L, Devevre E, Corthesy P, Romero P, Rufer N, and Speiser DE. Dominant human CD8 T-cell clonotypes persist simultaneously as memory and effector cells in memory phase. J Immunol. 182(11):6718-26 (2009)

Bricard G, Cesson V, Devevre E, Bouzourene H, Barbey C, Rufer N, Im SJ, Alves PM, Martinet O, Halkic N, Cerottini J-C, Romero P, Porcelli S, MacDonald R, and Speiser DE. Enrichment of human CD4+ Va24/Vb11 iNKT cells in intrahepatic malignant tumors. J Immunol. 182(8):5140-51 (2009)

Derré L, Bruyninx M, Baumgaertner P, Ferber M, Schmid D, Leimgruber A, Zoete V, Romero P, Michielin O, Speiser DE, and Rufer N. Distinct sets of alphabeta TCRs confer similar recognition of tumor antigen NY-ESO-1157-165 by interacting with its central Met/Trp residues. Proc. Natl. Acad. U.S.A. 105(39):15010-15015 (2008)

Voelter V, Rufer N, Reynard S, Greub G, Brookes R, Guillaume P, Grosjean F, Fagerberg T, Michielin O, Rowland-Jones S, Pinilla C, Leyvraz S, Romero, P, and Appay V. Characterization of Melan-A reactive memory CD8+ T-cells in a healthy donor. Int Immunol. 20:1087-1096 (2008)

Voelter V, Pica A, Laurent J, Rimoldi D, Bouzourene H, Sajadi A, Matter M, Romero P, Rufer N, and Speiser DE. An unusual case of metastatic melanoma sensitive to chemotherapy and immunotherapy-with late immune escape in the brain. Cancer Immun. 8:6-9 (2008)

Speiser DE, Baumgaertner P, Voelter V, Devevre E, Barbey C, Romero P, and Rufer N. Unmodified self antigen triggers human CD8 T cells with stronger tumor reactivity than altered antigen. Proc. Natl. Acad. U.S.A. 105(10):3849-54 (2008)

Rebeaud F, Hailfinger S, Posevitz-Fejfar A, Tapernoux M, Moser R, Rueda D, Gaide O, Guzzardi M, Iancu EM, Rufer N, Fasel N, and Thome M. The proteolytic activity of Malt1 plays a key role in T-cell activation. Nature Immunol. 9(3):272-81 (2008)

Iancu EM, Speiser DE, and Rufer N. Assessing ageing of individual T lymphocytes: Mission impossible? Mech Ageing Dev. 129(1-2):67-78 (2008)

Appay V, Bosio A, Lokan S, Wiencek Y, Biervert C, Kusters D, Devevre E, Speiser DE, Romero P, Rufer N, and Leyvraz S. Sensitive gene expression profiling of human T cell subsets reveals parallel post-thymic differentiation for CD4+ and CD8+ lineages. J Immunol. 179(11):7406-14 (2007)

Jorritsma A, Gomez-Eerland R, Dokter M, van de Kasteele W, Zoet YM, Doxiadis II, Rufer N, Romero P, Morgan RA, Schumacher TN, Haanen JB. Selecting highly affine and well expressed TCRs for gene therapy of melanoma. Blood. 110(10):3564-72 (2007)

Thadikkaran, L., Menzel, O., Tissot, J-D., Rufer, N. Proteomic and transcriptomic analysis of human CD8+ T lymphocytes over-expressing telomerase. Proteomics. (2007)

Romero, P., Zippelius, A., Kurth, I., Pittet, M.J., Touvrey, C., Iancu, E.M., Corthesy, P., Devevre, E., Speiser, D.E., Rufer, N. Four functionally distinct populations of human effector-memory CD8+ T lymphocytes. J Immunol. 178(7):4112-9 (2007)

Appay, V., Voelter, V., Rufer, N., Reynard, S., Jandus, C., Gasparini, D., Lienard, D., Speiser, D.E., Schneider, P., Cerottini, J.C., Romero, P., Leyvraz, S. Combination of transient lymphodepletion with busulfan and fludarabine and peptide vaccination in a phase I clinical trial for patients with advanced melanoma . J Immunother. 30(2):240-50 (2007)

Derré, L., Bruyninx, M., Baumgaertner, P., Devevre, E., Corthesy, P., Touvrey, C., Mahnke, Y.D., Pircher, H., Voelter, V., Romero, P., Speiser, D.E., Rufer, N. J. In Vivo Persistence of Codominant Human CD8+ T Cell Clonotypes Is Not Limited by Replicative Senescence or Functional Alteration. Immunol. 179(4):2368-79 (2007)

Barbey, C., Baumgaertner, P., Devevre, E., Rubio-Godoy, V., Derré, L., Bricard, G., Guillaume, P., Luescher, I.F., Liénard, D., Cerottini, J-C., Romero, P., Rufer, N., and Speiser, D.E. IL-12 controls cytotoxicity of a novel subset of self antigen-specific human CD28+ cytolytic T cells. J Immunol. 178(6):3566-74 (2007)

Tabiasco, J., Devevre, E., Rufer, N., Salaun, B., Cerottini, J-C., Speiser, D.E., and Romero, P. Human effector CD8+ T lymphocytes express toll like receptor-3 as a functional coreceptor. J Immunol. 177(12):8708-13 (2006)

Appay, V., Voelter, V., Rufer, N., Reynard, S., Gasparini, D., Liénard, D., Speiser, D.E., Schneider, P., Cerottini, J.C., Romero, P., and Leyvraz, S. Combination of transient lymphodepletion with Busulfan and Fludarabine and peptide vaccination in a phase I clinical trial for patients with advanced melanoma. J Immunother. 30(2):240-50 (2007)

Bianchi, T., Rufer, N., MacDonald, HR., and Migliaccio, M. The tumor suppressor p16(Ink4a) regulates T lymphocyte survival. Oncogene. 25(29):4110-5 (2006)

Colombetti, S., Fagerberg, T., Baumgaertner, P., Chapatte, L., Speiser, D.E., Rufer, N., Michielin, O., and Lévy, F. Impact of orthologous Melan-A peptide vaccination on the anti-self Melan-A/HLA-A2 T cell cross-reactivity. J Immunol. 176(11):6560-7 (2006)

Menzel, O., Migliaccio, M., Goldstein, D.R., Dahoun, S., Delorenzi, M., and Rufer, N. Mechanisms regulating the proliferative potential of human CD8+ T lymphocytes over-expressing telomerase. J Immunol. 177(6):3657-68 (2006)

Appay, V., Jandus, C., Voelter, V., Reynard, S., Coupland, S.E., Rimoldi, D., Lienard, D., Guillaume, P., Krieg, A.M., Cerottini, J.-C., Romero, P., Leyvraz, S., Rufer, N., Speiser, D.E. New generation vaccine induces effective melanoma-specific CD8+ T cells in the circulation but not in the tumor site, J Immunol., 177(3):1670-8 (2006)

Speiser, D.E., Baumgaertner, P., Barbey, C., Rubio-Godoy, V., Moulin, A., Corthesy, P., Devevre, E., Dietrich, P.Y., Rimoldi, D., Lienard, D., Cerottini, J.-C., Romero, P., Rufer, N. A novel approach to characterize clonality and differentiation of human melanoma-specific T cell responses: spontaneous priming and efficient boosting by vaccination, J Immunol, 177(2):1338-48 (2006)

Baumgaertner, P., Rufer, N., Devevre, E., Derré, L., Rimoldi, D., Geldhof, C., Voelter, V., Lienard, D., Romero, P., Speiser, D.E. Ex vivo detectable human CD8 T-cell responses to cancer-testis antigens, Cancer Res., 66(4):1912-6 (2006)

Appay, V., Speiser, D.E., Rufer, N., Reynard, S., Barbey, C., Cerottini, J.-C., Leyvraz, S., Pinilla, C., Romero, P. Decreased specific CD8(+) T cell cross-reactivity of antigen recognition following vaccination with Melan-A peptide, Eur J Immunol., 36(7):1805-14 (2006)

Bianchi, T., Rufer, N., MacDonald, H.R., Migliaccio, M. The tumor suppressor p16Ink4a regulates T lymphocyte survival, Oncogene, 25(29):4110-5 (2006)

Colombetti, S., Fagerberg, T., Baumgartner, P., Chapatte, L., Speiser, D.E., Rufer, N., Michielin, O., Lévy, F. Impact of orthologous melan-A peptide immunizations on the anti-self melan-A/HLA-A2 T cell cross-reactivity, J Immunol., 176(11):6560-7 (2006)

Migliaccio, M., Alves, PM., Romero, P., and Rufer, N. Distinct mechanisms control human naive and antigen-experienced CD8+ T lymphocyte proliferation. J Immunol. 176(4):2173-82 (2006)

Montes, M., Rufer, N., Appay, V., Reynard, S., Pittet, MJ., Speiser, DE., Guillaume, P., Cerottini, J.C., Romero, P., and Leyvraz, S. Optimum in vitro expansion of human antigen-specific CD8 T cells for adoptive transfer therapy. Clin Exp Immunol. 142, 292-302 (2005)

Le Gal, F.A., Ayyoub, M., Dutoit, V., Widmer, V., Jager, E., Cerottini, J.C., Dietrich, P.Y., Valmori, D. Distinct structural TCR repertoires in naturally occurring versus vaccine-induced CD8+ T-cell responses to the tumor-specific antigen NY-ESO-1. Journal of Immunotherapy, 28, 252-7 (2005)

Speiser, D.E., Lienard, D., Rufer, N., Rubio-Godoy, V., Rimoldi, D., Lejeune, F., Krieg, A.M., Cerottini, J.C., Romero, P. Rapid and strong human CD8(+) T cell responses to vaccination with peptide, IFA, and CpG oligodeoxynucleotide 7909. Journal of Clinical Investigation, 115, 739-46 (2005)

Migliaccio, M., Raj, K., Menzel, O., Rufer, N. Mechanisms That Limit the In Vitro Proliferative Potential of Human CD8+ T Lymphocytes. The Journal of Immunology, 174, 3335-3343 (2005)

Rufer, N., Reichenbach, P., Romero, P. Methods for the ex vivo characterization of human CD8+ T subsets based on gene expression and replicative history analysis. Methods in Molecular Medicine 109, 265-84 (2005)

Zippelius, A., Bioley, G., Le Gal, F.A., Rufer, N., Brandes, M., Batard, P., De Smedt, M., Plum, J., Speiser, D.E., Cerottini, J.C., Dietrich, P.Y., Romero, P., Pittet, M.J. Human thymus exports naive CD8 T cells that can home to nonlymphoid tissues. The Journal of Immunology 172: 2773-2777 (2004)

Zippelius, A., Batard, P., Rubio-Godoy, V., Bioley, G., Lienard, D., Lejeune, F., Rimoldi, D., Guillaume, P., Meidenbauer, N., Mackensen, A., Rufer, N., Lubenow, N., Speiser, D., Cerottini, J.C., Romero, P., Pittet, M.J. Effector function of human tumor-specific CD8 T cells in melanoma lesions: a state of local functional tolerance. Cancer Research 64, 2865-2873 (2004)

Papagno, L., Spina, C.A., Marchant, A., Salio, M., Rufer, N., Little, S., Dong, T., Chesney, G., Waters, A., Easterbrook, P., Dunbar, P.R., Sheperd, D., et al. CD8+ differentiation towards senescence following elevated activation in HIV-1 infection. Public Library of Science Biology 2, E20 (2003)

Dietrich, P-Y., Le Gal, F-A., Dutoit, V., Pittet, M.J., Trautman, L., Zippelius, A., Cognet, I., Widmer, V., Walker, P., Michielin, O., Guillaume, P., Connerotte, T., et al. Prevalent role of TCR alpha-chain in the selection of the preimmune repertoire specific for a human tumor-associated self-antigen. Journal of Immunology 170, 5103-5109 (2003)

Rufer N., Zippelius A., Batard P., Pittet M.J., Kurth I., Corthesy P., Cerottini J-C., Leyvraz S., Roosnek E., Nabholz M., Romero P. Identification of human pre-effector CD8+ T lymphocytes with distinct replicative history and partial effector functions. Blood 102, 1779-1787 (2003)

Sauce, D., Rufer, N., Mercier, P., Bodinier, M., Rémy-Martin, J-P., Duperrier, A., Ferrand, C., Hervé, P., Romero, P., Lang, F., Tiberghien, P. and Robinet, E. Retrovirus-mediated gene transfer in polyclonal T cells results in lower apoptosis and enhanced ex vivo expansion of CMV-reactive CD8 T cells as compared to EBV-reactive CD8 T cells. Blood 102, 1241-1248 (2003)

Reviews

Rufer, N. Molecular tracking of antigen-specific T-cell clones during immune responses. Curr Opin Immunol. 17, 441-447 (2005)

Rufer, N., Nabholz, M. Télomérase, élixir de jouvence des cellules humaines ? Médecine / Science 19, 345-350 (2003)

Romero, P., Valmori, D., Pittet, M.J., Zippelius, A., Rimoldi, D., Lévy, F., Dutoit, V., Rubio-Godoy, V., Michielin, O., Guillaume, P., Batard, P., Luescher, I.F., et al., Antigenicity and immunogenicity of Melan-A/Mart-1 derived peptides as targets for tumor reactive CTL in human melanoma. Immunology Reviews 188, 81-96 (2002)

Book chapter

Rufer, N., Reichenbach, P., and Romero, P. book chapter: Methods for the ex-vivo characterization of human CD8+ T subsets based on gene expression and replicative history analysis. Methods Mol Med. 109, 265-284. Adoptive Immunotherapy: Methods and Protocols. Edited by Ludewig B, and Hoffman, MW. Humana Press Inc.

List of publications Prof. Speiser

Published papers with peer reviews

Bricard, G., Cesson, V., Devevre, E., Bouzourene, H., Barbey, C., Rufer, N., Im, J. S., Alves, P. M., Martinet, O., Halkic, N., Cerottini, J. C., Romero, P., Porcelli, S. A., Macdonald, H. R. and Speiser, D. E., Enrichment of human CD4+ V(alpha)24/Vbeta11 invariant NKT cells in intrahepatic malignant tumors. J Immunol. 182: 5140-5151 (2009)

Iancu, E. M., Corthesy, P., Baumgaertner, P., Devevre, E., Voelter, V., Romero, P., Speiser, D. E. and Rufer, N., Clonotype selection and composition of human CD8 T cells specific for persistent herpes viruses varies with differentiation but is stable over time. J Immunol. 183: 319-331 (2009)

Lienard, D., Avril, M. F., Gal, F. A., Baumgaertner, P., Vermeulen, W., Blom, A., Geldhof, C., Rimoldi, D., Pagliusi, S., Romero, P., Dietrich, P. Y., Corvaia, N. and Speiser, D. E., Vaccination of Melanoma Patients With Melan-A/Mart-1 Peptide and Klebsiella Outer Membrane Protein P40 as an Adjuvant. J Immunother (2009)

Touvrey, C., Derre, L., Devevre, E., Corthesy, P., Romero, P., Rufer, N. and Speiser, D. E., Dominant human CD8 T cell clonotypes persist simultaneously as memory and effector cells in memory phase. J Immunol. 182: 6718-6726 (2009)

Alves, P. M., Levy, N., Stevenson, B. J., Bouzourene, H., Theiler, G., Bricard, G., Viatte, S., Ayyoub, M., Vuilleumier, H., Givel, J. C., Rimoldi, D., Speiser, D. E., Jongeneel, C. V., Romero, P. J. and Levy, F., Identification of tumor-associated antigens by large-scale analysis of genes expressed in human colorectal cancer. Cancer Immun. 8: 11 (2008)

Derre, L., Bruyninx, M., Baumgaertner, P., Ferber, M., Schmid, D., Leimgruber, A., Zoete, V., Romero, P., Michielin, O., Rufer, N. and Speiser, D. E., Distinct sets of alphabeta TCRs confer similar recognition of tumor antigen NY-ESO-1157-165 by interacting with its central Met/Trp residues. Proc Natl Acad Sci U S A. 105: 15010-15015 (2008)

Jandus, C., Bioley, G., Rivals, J. P., Dudler, J., Speiser, D. and Romero, P., Increased numbers of circulating polyfunctional Th17 memory cells in patients with seronegative spondylarthritides. Arthritis Rheum. 58: 2307-2317 (2008)

Jandus, C., Bioley, G., Speiser, D. E. and Romero, P., Selective accumulation of differentiated FOXP3(+) CD4 (+) T cells in metastatic tumor lesions from melanoma patients compared to peripheral blood. Cancer Immunol Immunother. 57: 1795-1805 (2008)

Schmitt, M., Schmitt, A., Rojewski, M. T., Chen, J., Giannopoulos, K., Fei, F., Yu, Y., Gotz, M., Heyduk, M., Ritter, G., Speiser, D. E., Gnjatic, S., Guillaume, P., Ringhoffer, M., Schlenk, R. F., Liebisch, P., Bunjes, D., Shiku, H., Dohner, H. and Greiner, J., RHAMM-R3 peptide vaccination in patients with acute myeloid leukemia, myelodysplastic syndrome, and multiple myeloma elicits immunologic and clinical responses. Blood. 111: 1357-1365 (2008)

Speiser, D. E., Baumgaertner, P., Voelter, V., Devevre, E., Barbey, C., Rufer, N. and Romero, P., Unmodified self antigen triggers human CD8 T cells with stronger tumor reactivity than altered antigen. Proc Natl Acad Sci U S A. 105: 3849-3854 (2008)

Voelter, V., Pica, A., Laurent, J., Rimoldi, D., Bouzourene, H., Sajadi, A., Matter, M., Romero, P., Rufer, N. and Speiser, D. E., An unusual case of metastatic melanoma sensitive to chemotherapy and immunotherapy, with late immune escape in the brain. Cancer Immun. 8: 6 (2008)

Alves, P.M., Viatte, S., Fagerberg, T., Michielin, O., Bricard, G., Bouzourene, H., Vuilleumier, H., Kruger, T., Givel, J.C., Levy, F., Speiser, D.E., Cerottini, J.C., Romero, P. Immunogenicity of the carcinoembryonic antigen derived peptide 694 in HLA-A2 healthy donors and colorectal carcinoma patients. Cancer Immunol Immunother. Apr 20; [Epub ahead of print] (2007)

Le Gal, F.A., Widmer, V.M., Dutoit, V., Rubio-Godoy, V., Schrenzel, J., Walker, P.R., Romero, P.J., Valmori, D., Speiser, D.E., Dietrich, P.Y. Tissue homing and persistence of defined antigen-specific CD8+ tumor-reactive T-cell clones in long-term melanoma survivors. J Invest Dermatol. 127(3):622-9 (2007)

Alves, P., Lévy, N., Bouzourene, H., Viatte, S., Bricard, G., Ayyoub, M., Vuilleumier, H., Givel, J.-C., Halkic, N., Speiser, D., Romero, P., Lévy, F. Molecular and immunological evaluation of the expression of cancer/testis gene products in human colorectal cancer. Cancer Immunol Immunother, 56(6):839-47 (2007)

Tabiasco, J., Devevre, E., Rufer, N., Salaun, B., Cerottini, J.C., Speiser, D., Romero, P. Human effector CD8+ T lymphocytes express TLR3 as a functional coreceptor. J Immunol. 177(12):8708-13 (2006)

Batard, P., Peterson, D.A., Devevre, E., Guillaume, P., Cerottini, J.-C., Rimoldi, D., Speiser, D.E., Winther, L. and Romero, P. Dextramers: new generation of fluorescent MHC class I / peptide multimers for visualization of antigen-specific CD8 T cells. J Immunol Methods 310: 136-48 (2006)

Guillaume, P., Baumgaertner, P., Angelov, G.S., Speiser, D., and Luescher, I.F. Fluorescence-activated cell sorting and cloning of bona fide CD8+ CTL with reversible MHC-peptide and antibody Fab' conjugates. J Immunol. 177:3903-3912 (2006)

Bioley G., Jandus, C., Tuyaerts, S., Rimoldi, D., Kwok, W.W., Speiser, D.E., Tiercy, J.-M., Thielemans, K., Cerottini, J.-C. and Romero, P. Melan-A / MART-1 specific CD4 T cells in melanoma patients: identification of new epitopes and ex vivo visualization of specific T cells by MHC class II tetramers. J Immunol. 177: 6769-6779 (2006)

Colombetti, S., Fagerberg, T., Baumgaertner, P., Chapatte, L., Speiser, D.E., Rufer, N., Michielin, O., Lévy, F. Impact of orthologous melan-A peptide immunizations on the anti-self melan-A/HLA-A2 T cell cross-reactivity, J Immunol., 176(11):6560-7 (2006)

Pittet, M.J., Gati, A., Le Gal, F.A., Bioley, G., Guillaume, P., de Smedt, M., Plum, J., Speiser, D.E., Cerottini, J.-C., Dietrich, P.Y., Romero, P., Zippelius, A. Ex vivo characterization of allo-MHC-restricted T cells specific for a single MHC-peptide complex, J Immunol., 176(4):2330-6 (2006)

Appay, V., Reynard, S., Voelter, V., Romero, P., Speiser, D.E., Leyvraz, S. Immuno-monitoring of CD8+ T cells in whole blood versus PBMC samples. J Immunol Methods, 309(1-2):192-9 (2006)

Michielin, O., Blanchet, J.S., Fagerberg, T., Valmori, D., Rubio-Godoy, V., Speiser, D., Ayyoub, M., Alves, P., Luescher, I., Gairin, J.E., Cerottini ,J.C., Romero, P. Tinkering with nature: The tale of optimizing peptide based cancer vaccines. Review Article. Cancer Treat Res. 123:267-291 (2005)

Speiser, D.E. Tumor specific T cells induced in melanoma patients either naturally or by vaccination with peptides, IFA ± CpG oligodeoxynucleotides. Cancer Immunity; 5 Suppl 1:23 (2005) http://www.cancerimmunity.org/v5suppl1p23/041244_abs.htm

Ayyoub, M., Merlo, A., Hesdorffer, C.S., Speiser, D., Rimoldi, D., Cerottini, J.C., Ritter, G., Chen, Y.T., Old, L.J., Stevanovic, S., Valmori, D., Distinct but overlapping T helper epitopes in the 37-58 region of SSX-2. Clinical Immunology. 114:70-78 (2005)

Bricard, G., Bouzourene, H., Martinet, O., Rimoldi, D., Halkic, N., Gillet, M., Chaubert, P., Macdonald, H.R., Romero, P., Cerottini, J.C., Speiser, D.E., Naturally acquired MAGE-A10 and SSX-2 specific CD8⁺ T cell responses in patients with hepatocellular carcinoma. Journal of Immunology. 174:1709-1716 (2005)

Speiser, D.E., Cerottini, J.C., Romero, P., Tumor cell recognition efficiency by T cells. PLoS Medicine 29:2 (3) e77 (2005)

Speiser D.E. and Romero, P., Toward improved immunocompetence of adoptively transferred CD8+ T cells,. Journal of Clinical Investigation, 115:1467-1469 (2005)

Speiser, D.E., Immunological Techniques: Ex vivo characterization of T cell mediated immune responses in cancer. Editorial Overview. Current Opinion in Immunology. 17:419-422 (2005)

Montes, M., Rufer N., Appay V., Reynard S., Pittet M.J., Speiser D.E., Guillaume P., Cerottini J.C., Romero P., Leyvraz S. Optimal in vitro expansion of human antigen specific CD8+ T cells for adoptive transfer therapy. Clinical and Experimental Immunology, 142.292-302 (2005)

Ayyoub, M., Hesdorffer, C.S., Metthez, G., Stevanovic, S., Ritter, G., Chen, Y.T., Old, L.J., Speiser, D., Cerottini, J.C., Valmori, D., Identification of an SSX-2 epitope presented by dendritic cells to circulating autologous CD4+ T cells. Journal of Immunology, 172 7206-7211 (2004)

Ayyoub, M., Hesdorffer, C.S., Montes, M., Merlo, A., Speiser, D., Rimoldi, D., Cerottini, J.C., Ritter, G., Scanlan, M., Old, L.J., Valmori, D. An immunodominant SSX-2-derived epitope recognized by CD4+ T cells in association with HLA-DR. Journal of Clinical Investigation 113: 1225-33 (2004)

Rothenfusser, S., Hornung, V., Ayyoub, M., Britsch, S., Towarowski, A., Krug, A., Sarris, A., Lubenow, N., Speiser, D., Endres, S., Hartmann, G. CpG-A and CpG-B oligonucleotides differentially enhance human peptide-specific primary and memory CD8+ T-cell responses in vitro. Blood 15, 2162-9 (2004)

Speiser, D.E., Pittet, M.J., Guillaume, P., Lubenow, N., Hoffman, E., Cerottini, J.C., Romero, P. Ex vivo analysis of human antigen-specific CD8+ T-cell responses, quality assessment of fluorescent HLA-A2 multimer and interferon-gamma ELISPOT assays for patient immune monitoring. Journal of Immunotherapy 27, 298-308 (2004)

Lienard, D., Rimoldi, D., Marchand, M., Dietrich, P.Y., van Baren, N., Geldhof, C., Batard, P., Guillaume, P., Ayyoub, M., Pittet, M.J., Zippelius, A., Fleischhauer, K., Lejeune, F., Cerottini, J.C., Romero, P., Speiser, D.E. Ex vivo detectable activation of Melan-A-specific T cells correlating with inflammatory skin reactions in melanoma patients vaccinated with peptides in IFA. Cancer Immunity 4, 4 (2004)

Zippelius A., Bioley G., Le Gal F-A., Rufer N., Brandes M., Batard P., De Smedt M., Plum J., Speiser D.E., Cerottini J-C., Dietrich P-Y., Romero P., Pittet M.J. Targeting of secretory IgA to Peyer's patch dendritic and T cells after transport by intestinal M cells. Journal of Immunology 172, 2773-7 (2004)

Zippelius, A., Batard, P., Rubio-Godoy, V., Bioley, G., Lienard, D., Lejeune, F., Rimoldi, D., Guillaume, P., Meidenbauer, N., Mackensen, A., Rufer, N., Lubenow, N., Speiser, D., Cerottini, J.C., Romero, P., Pittet, M.J. Effector function of human tumor-specific CD8 T cells in melanoma lesions, a state of local functional tolerance. Cancer Research 64, 2865-73 (2004)

Valmori, D., Dutoit, V., Ayyoub, M., Rimoldi, D., Guillaume, P., Liénard, D., Lejeune, F., Cerottini, J.C., Romero, P., Speiser, D.E. Simultaneous CD8+ T cell responses to multiple tumor antigen epitopes in a multipeptide melanoma vaccine. Cancer Immunity 3, 15 (2003)

Ayyoub, M., Rimoldi, D., Guillaume, P., Romero, P., Cerottini, J.-C., Valmori, D. Speiser, D. E. Tumor reactive SSX-2 specific CTL are selectively expanded during spontaneous responses to antigen expressing tumors in melanoma patients. Cancer Research 63, 5601-6 (2003)

Pittet, M.J., Rubio-Godoy, V., Bioley, G., Guillaume, P., Batard, P., Speiser, D.E., Luescher, I., Cerottini, J.C., Romero, P., Zippelius, A. ά-3 domain mutants of peptide/MHC multimers allow the selective isolation of high avidity tumor-reactive CD8 T cells for adoptive T cell therapy. Journal Immunology 171, 1844-1849 (2003)

Reviews

Jandus, C., Speiser, D. and Romero, P., Recent advances and hurdles in melanoma immunotherapy. Pigment Cell Melanoma Res (2009)

Iancu, E. M., Speiser, D. E. and Rufer, N., Assessing ageing of individual T lymphocytes: mission impossible? Mech Ageing Dev. 129: 67-78 (2008)

Michielin, O., Rufer, N., Romero, P., Laurent, J., Cerottini, J. P., Gugisberg, D., Leyvraz, S. and Speiser, D., New developments in cancer immunotherapy. Rev Med Suisse. 4: 1248-1251 (2008)

Lejeune, F. J., Rimoldi, D. and Speiser, D., New approaches in metastatic melanoma: biological and molecular targeted therapies. Expert Rev Anticancer Ther. 7: 701-713 (2007)

Speiser, D.E., Cerottini, J.-C. and Romero, P. The human T cell response to melanoma antigens. Adv Immunol. 92: 187-224 (2006)

Mahnke, Y., Speiser, D., Luescher, I.F., Cerottini, J.C., Romero, P., Recent advances in tumour antigen-specific therapy: in vivo veritas. International Journal of Cancer, 113: 173-178 (2005)

Speiser, D.E., Immunological techniques: ex vivo characterization of T cell-mediated immune responses in cancer. Current Opinion in Immunology; 17:419-22 (2005)

Slingluff, C.L. Jr., Speiser, D.E., Progress and controversies in developing cancer vaccines. Journal of Translational Research. 3:18 (2005)

Romero, P., Cerottini, J.C., Speiser, D.E., Monitoring tumor antigen specific T-cell responses in cancer patients and phase I clinical trials of peptide-based vaccination. Cancer Immunol Immunother. 53, 249-55 (2004)

Romero, P., Valmori, D., Pittet, M.J., Zippelius, A., Rimoldi, D., Lévy, F., Dutoit, V., Rubio-Godoy, V., Michielin, O., Guillaume, P., Batard, P., Luescher, I.F., Lejeune, F., Liénard, D., Rufer, N., Dietrich, P.Y., Speiser, D.E., Cerottini, J.C. Antigenicity and immunogenicity of Melan-A/MART-1 derived peptides as targets for tumor reactive CTL in human melanoma. Immunological Reviews 188, 81-96 (2002)

National Center of Competence in Research - NCCR Molecular Oncology