T cell response to Hu-D peptides in patients with anti-Hu syndrome Laboraatory Investigation DOI:
Cite this article as: Rousseau, A., Benyahia, B., Dalmau, J. et al. J Neurooncol (2005) 71: 231. doi:10.1007/s11060-004-1723-1 Abstract
The anti-Hu syndrome is the most common paraneoplastic neurologic syndrome but the exact mechanism of immune mediated neuronal injury remains unknown. Anti-Hu antibodies do not appear to play a pivotal role in the pathogenesis of the disease. To assess cell-mediated immunity, we selected 51 peptides from the Hu-D sequence and tested their ability to bind to six common HLA class I molecules. Stable complexes with purified HLA molecules were obtained with 19/51 (37%) selected peptides. Subsequently, the ability of the 19 HLA-binding peptides to stimulate T cells from 10 patients and 10 control subjects was evaluated by detecting IFN-γ secretion. An anti-peptide T-cell response was observed in 7/10 Hu-positive patients but also in 3/10 control subjects. Overall, a significant T-cell activation occurred in response to 74% (14 out of 19) of the selected peptides in the Hu-positive patients vs. 16% (3 out of 19) in the control group (
p < 0.001). In addition, T cells of patients tested within 3 months of the onset of anti-Hu syndrome responded to 82% (14 out of 17) of assessed Hu-D peptides vs. 37% (7 out of 19) in patients tested 1 year or more after developing the syndrome ( p < 0.01). Thus, the present study suggests a role of cellular immunity during the course of anti-Hu syndrome. Keywords anti-Hu syndrome cellular immunity ELISPOT assay HLA/peptide complexes Hu-Depitopes IFN-γ paraneoplastic disorders T lymphocytes References Darnell, RB, Posner, JB 2003 Paraneoplastic syndromes involving the nervous system N Engl J Med 349 1543 1554 CrossRef PubMed Google Scholar Altermatt, HJ, Rodriguez, M, Scheithauer, BW, Lennon, VA 1991 Paraneoplastic anti-Purkinje and type 1 anti-neuronal nuclear auto-antibodies bind selectively to central, peripheral, and autonomic nervous system cells Lab Invest 65 412 420 Google Scholar Manley, GT, Sillevis Smitt, P, Dalmau, J, Posner, JB 1995 Hu antigens: reactivity with Hu antibodies, tumor expression, and major immunogenic sites Ann Neurol 38 102 110 Google Scholar Carpentier, AF, Voltz, R, DesChamps, T, Posner, JB, Dalmau, J, Rosenfeld, MR 1998 Absence of HuD gene mutations in paraneoplastic small cell lung cancer tissue Neurology 50 1919 Google Scholar Greenlee, JE, Parks, NT, Jaeckle, KA 1993 Type 2a (‘anti-Hu’) antineuronal antibodies produce destruction of rat cerebellar granule neurons in vitro Neurology 43 2049 2054 Google Scholar Schäfer, KH, Klotz, M, Mergner, M, Mestres, P, Schimrigk, K, Blaes, F 2000 IgG-mediated cytotoxicity to myenteric plexus cultures in patients with paraneoplastic neurological syndromes J Autoimmun 15 479 484 Google Scholar Sillevis Smitt, PAE, Manley, GT, Posner, JB 1995 Immunization with the paraneoplastic encephalomyelitis antigen HuD does not cause neurologic disease in mice Neurology 45 1873 1878 Google Scholar Carpentier, AF, Rosenfeld, MR, Delattre, JY, Whalen, RG, Posner, JB, Dalmau, J 1998 DNA vaccination with HuD inhibits growth of a neuroblastoma in mice Clin Cancer Res 4 2819 2824 Google Scholar Graus, F, Ribalta, T, Campo, E, Monforte, R, Urbano, A, Rozman, C 1990 Immunohistochemical analysis of the immune reaction in the nervous system in paraneoplastic encephalomyelitis Neurology 40 219 222 Google Scholar Voltz, R, Dalmau, J, Posner, JB, Rosenfeld, MR 1998 T-cell receptor analysis in anti-Hu associated paraneoplastic encephalomyelitis Neurology 51 1146 1150 Google Scholar Wanschitz, J, Hainfellner, JA, Kristoferitsch, W, Drlicek, M, Budka, H 1997 Ganglionitis in paraneoplastic subacute sensory neuropathy: a morphologic study Neurology 49 1156 1159 Google Scholar Plonquet, A, Gherardi, RK, Créange, A, Antoine, JC, Benyahia, B, Grisold, W, Drlicek, M, Dreyfus, P, Honnorat, J, Khouatra, C, Rouard, H, Authier, FJ, Farcet, JP, Delattre, JY, Delfau-Larue, MH 2002 Oligoclonal T-cells in blood and target tissues of patients with anti-Hu syndrome J Neuroimmunol 122 100 105 Google Scholar Benyahia, B, Liblau, R, Merle-Beral, H, Tourani, JM, Dalmau, J, Delattre, JY 1999 Cell-mediated autoimmunity in paraneoplastic neurologic syndromes with anti-Hu antibodies Ann Neurol 45 162 167 Google Scholar Tanaka, K, Tanaka, M, Inuzuka, T, Nakano, R, Tsuji, S 1999 Cytotoxic T lymphocyte-mediated cell death in paraneoplastic sensory neuronopathy with anti-Hu antibody J Neurol Sci 163 159 162 Google Scholar Tanaka, M, Maruyama, Y, Sugie, M, Motizuki, H, Kamakura, K, Tanaka, K 2002 Cytotoxic T cell activity against peptides of Hu protein in anti-Hu syndrome J Neurol Sci 201 9 12 Google Scholar Plonquet, A, Garcia-Pons, F, Fernandez, E, Philippe, C, Marquet, J, Rouard, H, Delfau-Larue, MH, Kosmatopoulos, K, Lemonnier, F, Farcet, JP, Gherardi, RK, Langlade-Demoyen, P 2003 Peptides derived from the onconeural HuD protein can elicit cytotoxic responses in HHD mouse and human J Neuroimmunol 142 93 100 Google Scholar Terasaki, PI, McClelland, JD 1964 Microdroplet assay of human serum cytotoxins Nature 204 998 1000 Google Scholar Choppin, J, Martinon, F, Gomard, E, Barhaoui, E, Connan, F, Bouillot, M, Lévy, JP 1990 Analysis of physical interaction between peptides and HLA molecules and application to the detection of HIV-1 antigenic peptides J Exp Med 172 889 899 Google Scholar Connan, F, Hlavac, F, Hoebeke, J, Guillet, JG, Choppin, J 1994 A simple assay for detection of peptides promoting the assembly of HLA class I molecules Eur J Immunol 24 777 780 Google Scholar Subklewe, M, Chahroudi, A, Bickham, K, Larsson, M, Kurilla, MG, Bhardwaj, N, Steinman, RM 1999 Presentation of Epstein-Barr virus latency antigens to CD8+, interferon-γ-secreting, T lymphocytes Eur J Immunol 29 3995 4001 Google Scholar Choppin, J, Cohen, W, Bianco, A, Briand, JP, Connan, F, Dalod, M, Guillet, JG 2001 Characteristics of HIV-1 Nef regions containing multiple CD8+ T cell epitopes: wealth of HLA-binding motifs and sensitivity to proteasome degradation J Immunol 166 6164 6169 Google Scholar Lau, LL, Jamieson, BD, Somasundaram, T, Ahmed, R 1994 Cytotoxic T-cell memory without antigen Nature 369 648 652 Google Scholar Ahmed, R, Gray, D 1996 Immunological memory and protective immunity: understanding their relation Science 272 54 60 Google Scholar Dalmau, J, Graus, F, Cheung, NKV, Rosenblum, MK, Ho, A, Canete, A, Delattre, JY, Thompson, SJ, Posner, JB 1995 Major histocompatibility proteins, anti-Hu antibodies, and paraneoplastic encephalomyelitis in neuroblastoma and small cell lung cancer Cancer 75 99 109 Google Scholar Scheibenbogen, S, Lee, KH, Stevanovic, S, Witzens, M, Willhauck, M, Waldmann, V, Naeher, H, Rammensee, HG, Keilholz, U 1997 Analysis of the T cell response to tumor and viral peptide antigens by an IFN-γ-ELISPOT assay Int J Cancer 71 932 936 Google Scholar W, Herr, J, Schneider, AW, Lohse, KH, Meyer zum Buschenfelde, T, Wolfel 1996 Detection and quantification of blood-derived CD8+ T lymphocytes secreting tumor necrosis factor alpha in response to HLA-A2.1-binding melanoma and viral peptide antigens J Immunol Meth 191 131 142 Google Scholar Marincola, FM, Rivoltini, L, Salgaller, ML, Player, M, Rosenberg, SA 1996 Differential anti-MART-1/Melan-A CTL activity in peripheral blood of HLA-A2 melanoma patients in comparison to healthy donors: evidence of in vivo priming by tumor cells J Immunother Emphasis Tumor Immunol 19 266 277 Google Scholar Loftus, D, Castelli, C, Clay, TM, Squarcina, P, Marincola, FM, Nishimura, MI, Parmiani, G, Appella, E, Rivoltini, L 1996 Identification of epitope mimics recognized by CTL reactive to the melanoma/melanocyte-derived peptide MART-1(27–35) J Exp Med 184 647 657 Google Scholar Bach, JF, Koutoulov, S, Endert, PM 1998 Are there unique autoantigens triggering autoimmune diseases Immunol Rev 164 139 155 Google Scholar Rammensee, HG, Friede, T, Stevanovic, S 1995 MHC ligands and peptide motifs: first listing Immunogenetics 50 201 212 Google Scholar Scheibenbogen, C, Lee, KH, Mayer, S, Stevanovic, S, Moebius, U, Herr, W, Rammensee, HG, Keilholz, U 1997 A sensitive ELISPOT assay for detection of CD8+ T lymphocytes specific for HLA class I-binding peptide epitopes derived from influenza proteins in the blood of healthy donors and melanoma patients Clin Cancer Res 3 221 226 Google Scholar
Henson RA, Urich H: Cancer and the Nervous System. The Neurological Manifestations of Systemic Malignant Disease. Blackwell Scientific Publications, New York, 1982.
Lalvani, A, Brookes, R, Hambleton, S, Britton, WJ, Hill, AVS, McMichael, AJ 1997 Rapid effector function in CD8+ memory T cells J Exp Med 186 859 865 Google Scholar