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T Cell Hybridomas: A Workshop at the Basel Institute for Immunology PDF

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Preview T Cell Hybridomas: A Workshop at the Basel Institute for Immunology

Current Topics in Microbiology 100 and Immunology Editors M. Cooper, Birmingham/Alabama· W. Henle, Philadelphia P.H. Hofschneider, Martinsried· H. Koprowski, Philadelphia F. Melchers, Basel· R. Rott, GieSen· H.G. Schweiger; LadenburglHeidelberg· P.K. Vogt, Los Angeles R. Zinkernagel, Zurich T Cell Hybridomas A Workshop at the Basel Institute for Immunology Organized and Edited by H. v. Boehmer, W. Haas, G. Kohler, F. Melchers and J. Zeuthen With the Collaboration of S. Buser-Boyd With 52 Figures Springer-Verlag Berlin Heidelberg New York 1982 Editors H. V. Boehmer, W. Haas G. Kohler, F. Melchers J. Zeuthen Basel Institute for Immunology Grenzacherstrasse 487 CH-4005 Basel Switzerland ISBN-13: 978-3-642-68588-0 e-ISBN-13: 978-3-642-68586-6 DOl: 10.1007/978-3-642-68586-6 This work is subject to copyright. All rights are reserved, whether the whole or part of the material is con- cerned, specially those of translation, reprinting, fe-use of illustration broadcasting, reproduction by pbotocopying machine or similar means, and storage in data banks. Under § 54 of the German Copyright Law where copies are made for other than private use a fee is payable to 'Verwertungsgesellschaft Wort'. Munich. © by Springer-Verlag Berlin Heidelberg 1982 Library of Congress Catalog Card Number 15-12910 Softcover reprint of the hardcover 1st edition 1982 The use of registered names, trademarks, etc. in this publication, does nor imply. even in the absence of a specific statement, that such names are exempt from the relevant protective laws and regulations and therefore free for general use. Typesetting: FotoSatz Pfeifer, Germering Printing and binding: Universitiitsdrnckerei H. StUrtz AG. Wiirzburg 2121/3321-543210 Table of Contents Preface . . . . . . . . . . . . . . . . . . . . . . . . .. VII K. Fischer Lindahl, A.A. Nordin, M.H. Schreier: Lectin- Dependent Cytolytic and Cytolymic T Helper Clones and Hybridomas . . . . . . . . . . . . . . . . . . . . . . . 1 Z. Eshhar, T. Waks, T . .oren, G. Berke, Y. Kaufmann: Cytotoxic T Cell Hybridomas: Generation and Characterization .................... 11 E.J. Culbert, S. Kontiainen, L.M. Douglas, M. Feldmann: Definition of Function-Related Isotypic Markers on TCells ........................ 19 M. Taniguchi, T. Tokuhisa, M. Kanno, T. Honjo: An Antigen-Specific Suppressor T Cell Factor Controlled by Two Genes in the Immunglobulin Heavy Chain Linkage Group and in the I-J Subregion ofthe H-2 Complex '" 33 M.J. Taussig, G.L. Asherson, A. Holliman, N. Richardson, G. Skibinski: Structural and Functional Studies on Antigen- Specific Suppressor Factors from T Cells and T Cell Hybrids. . . . . . . . . . . . . . . . . . . . . . . . .. 43 D.R. Webb, B.A. Araneo, C. Healy, J.A. Kapp, K. Krupen, I. Nowowiejski, C.W. Pierce, C.M. Sorensen, S. Stein, K.J. Wieder: Purification and Biochemical Analysis of Antigen- Specific Suppressor Factors Isolated from T-Cell Hybridomas . . . . . . . . . . . . . . . . . . . . . 53 M.E. Dorf, K. .okuda, M. Minami: Dissection of a Suppressor Cell Cascade .............. 61 M. Cramer, R. Mierau, T. Takemori, G. Suzuki, T. Tada: 4-Hydroxy-3-nitro-phenylacetyl (NP)-Specific T Cell Hybridomas . . . . . . . . . . . . . . . . . . . . . . 69 M. Suemura, K. Sugimura, A. Ishizaka, S. Kobatake,_ O. Shiho, H. Deguchi, S. Kishimoto, Y. Yamamura, T. Kishimoto: Regulation of the IgE Response by IgE Class-Specific Suppressor T Hybridomas . . . . . . . 79 P. Ricciardi-Castagnoli, F. Robbiati, E. Barbanti, G. Doria, L. Adorini: Establishment of Functional, Antigen-Specific T Cell Lines by RadLV -Induced Transformation of Murine T Lymphocytes . . . . . 89 P. Lonai, E. Arman, S. Bitton-Grossfeld, 1. Grooten, G. Hiimmerling: H-2-Restricted Helper Hybridomas: One Locus or Two Control Dual Specificity? ......... 97 Z. Eshhar, T. Waks, H. Zinger, E. Mozes: T Cell Hybri- domas Producing Antigen-Specific Factors Express Heavy- Chain~Variable-Region Determinants ......... 103 S. Carel, C. Bron, G. Corradin: Cytochrome f. Specific T Cell Hybrid . . . . . . . . . . . . . . . . . . . . .. 111 E. Heber-Katz, D. Hansburg, R.H. Schwartz: The Effect of Antigen Presentation on the Fine Specificity of Anti- Cytochrome f T Cell Hybridomas . . . . . . . . . . .. 117 A.M. Schmitt-Verhulst, M.A. Cooley, A. Guimezanes, C. De Preval, F. Albert, M. Buferne: Analysis of the Anti-Self + TNP Immune Response: T Cell Lines, Clones and Hybridomas. . . . . . . . . . . . . . . . 125 M.H. Schreier, R. Tees, L. Nordin, A.A. Nordin: Func- tional Characteristics of T Cell Hybridomas Obtained by Fusion of TCGF-Dependent Helper T Cell Clones with BW5147 . . . . . . . . . . . . . . . . . . . . . 135 K.-O. Gronvik, 1. Andersson, F. Melchers, 1. Zeuthen: A Stable TCGF-Producing T Cell Hybridoma and its Thioguanine-Resistant Variant Suitable as a Tool for the Construction of New Functional T Hybridomas . . . .. 143 F. Melchers, 1. Zeuthen, W. Gerhard: Influenza Virus- Specific Murine T Cell Hybridomas Which Recognize Virus Hemagglutinin in Conjunction with H-2d and Display Helper Functions for B Cells . . . . . . . . . . . . .. 153 1. Andersson, F. Melchers, 1. Zeuthen: A "Panreactive" T Cell Line and T Cell Hybridoma: Their Function in Helping B Cells . . . . . . . . . . . . . . . . . . . . 165 C. Corbel, 1. Andersson, F. Melchers, 1. Zeuthen: A "Panreactive" T Cell Hybridoma Which Produces TCGF Constitutively . . . . . . . . . . . . . . . . . . . . .. 173 A. Altman, R.D. Schreiber, D.H. Katz: Production of Antigen-Nonspecific Immunoregulatory Lymphokines by T Cell Hybridomas . . . . . . . . . . . . . . . . .. 179 E.C. DeFreitas, S. Vella, A. Linnenbach, C.M. Croce, H. Koprowski: Human T Cell Hybridomas with Tetanus- Toxoid-Specific Helper Activity ............ 191 K. Pfizenmaier, M. R611inghoff, H. Wagner: Functional Analysis of Il-2 Produced by T-Cell Hybridomas: Il-2 Promotes T-Cell Growth But Does Not Mediate T-Cell or B-Cell Maturation and Differentiation . . . . . . .. 203 S. Gillis, D. Mochizuki: Molecular Characterization of Interleukin 2 Produced from Tumor Cell Lines and T Cell Hybridomas . . . . . . . . . . . . . . . . . .. 211 J.W. Schrader, I. Clark-Lewis: The Use ofT Cell Hybri- domas in the Biochemical and Biological Characterization of Multiple Regulatory Factors Produced by T Cells. 221 C. Corbel: Different Factors Active in Lymphoid and Hematopoietic Proliferation Produced by Single Clones of Helper T Cell Hybridomas . . . . . . . . . . . . .. 231 N.H. Ruddle, B.S. Conta: Lymphotoxin and Immune (y) Interferon Production by T Cell Lines and Hybrids . . .. 239 J.F.A.P. Miller, P.L. Mottram, W.R. Thomas, J. McKimm- Breschkin, I.L. Walker, J. Gamble: Antigen Specificity of Continuous T Cell Lines 249 List of Contributors . . . . . 257 Indexed in Current Contents Preface For more than ten years cell fusion techniques have been applied in studies on various lymphocyte functions. Ig expression was first studied in hybrids obtained by fusing myeloma cells with fibroblasts (1) or lymphomas (2), both of which do not produce Ig, and with Ig- producing myelomas (3) or human blood lymphocytes (4). Kohler and Milstein (5) fused a myeloma with spleen cells from immunized mice. Up to 10% of the hybrids obtained secreted antibodies specific for the immunizing antigen. This suggested that plasma cells preferenti- ally fused with the myeloma cells, a finding which was of enormous practical value. It was found that both Band T lymphocytes could be fused with the T cell tumor BW5147, which is however not permissive for Ig synthesis (6). A very large number of T cell hybridomas were generated by fusing BW5147 with cell populations containing in vivo or in vitro activated cells (7). The hybrids showed no specific T cell functions and binding assays for T cell receptors were not available. In particular, no hybrids were obtained which expreS1ed specific cytolytic activity that could be tested in short-term Cr- release assays (8). However, the frustrations expressed about these failures, published in January, 1978 (9), were relieved by Taniguchi and Miller's publication a few months later of T cell hybridomas producing antigen-specific suppressor factors (10). Unfortunately, their hybrids rapidly lost factor production. Subsequently, many laboratories generated murine and human T cell hybridomas which produced antigen-specific molecules that suppressed or induced various lymphocyte functions (this volume; 8,11). Many T cell hybridomas could be induced to produce various lymphokines (this volume; 12). Some T cell hybrids expressed antigen receptors which could be identi- fied by binding antigen or antiidiotypic antibodies or antigen- induced lymphokine production (this volume; 13). Indeed, T cell hybridomas expressing specific lytic activity could also be generated (this volume; 14). It is remarkable that BW5147 is permissive for expression of all these T cell functions. Many more fusion experi- ments will be required - including inter species fusions - to determine the optimal conditions for fusion and expression of particular T cell functions in hybrid cells. Although the problem of stable expression of particular T cell functions in hybridomas has not yet been com- pletely solved, several laboratories have generated sufficient numbers of hybrid cells to allow purification and biochemical analysis of antigen-specific T cell factors. Most advanced is the analysis of suppressor factors (this volume). T cell hybridomas are not the only source of monoclonal T cell pro- ducts. First, several murine and human T cell tumors can be induced to express normal T cell functions such as lymphokine production (15). Second, transformed T cell lines expressing specific functions can be obtained by infection of mice or cells in vitro with radiation leukemia virus (16). Third, clones of all major T cell classes can IX now be grown continuously in tissue culture (17). To help evaluate the potential of T cell hybridomas for understanding the functioning of the immune system as well as for practical purposes, the workshop on "T Cell Hybridomas: Sources of Specific Mediators in the Immune System" was held January 27-29, 1982, at the Basel Insti- tute for Immunology. The techniques used, the difficulties encoun- tered and the present state of art were discussed, and most of this is presented in the following papers. References 1. Periman (1970) Nature 228: 1086; Coffino et al. (1971) Nature New Biol 231: 87; Bevan et al. (1972) Prog Biophys Mol Biol 25: 131 2. Mohit (1971) Proc Natl Acad Sci USA 68: 3045 3. Cotton and Milstein (1973) Nature 244: 42 4. Schwaber and Cohen (1974) Proc Natl Acad Sci USA 71: 2203 5. Kohler and Milstein (1975) Nature 256: 495 6. Iverson et al. (1978) Curr Top Microbiol Immunol 81: 192 7. Hammerling (1977) Eur J Immunol 7: 743; Goldsby et al. (1977) Nature 267: 707; Kohler et al. (1977) Eur J Immunol 7: 758 8. Curr Top Microbiol Immunol 81: 195-231 (1978) 9. Melchers (1978) Nature 271: 9 10. Taniguchi and Miller (1978) J Exp Med 148: 373 11. Research Monographs in Immunology, Vol. 3: Monoclonal Antibodies and T Cell Hybridomas: Perspectives and Technical Advances, G.J. Hammerling, U. Hammerling, J.F. Kearney, eds., Elsevier/ North Holland, 1982; Watanabe et al. (1978) J Immunol 121: - 2113; Hewitt and Liew (1979) Eur J Immunol 9: 572; Okada et al. (1981) Proc Natl Acad Sci USA 78: 7717; Hiramatsu et al. (1981) J Immunol 127: 1118; Irigoyen et al. (1981) J Exp Med 154: 1827 12. Harwell et al. (1981) J Exp Med 152: 893; Kappler et al. (1981) J Exp Med 153: 1198 13. Kappler et al. (1981) J Exp Med 153: 1198 14. Nabholz et al. (1981) Nature 287: 437 15. Behring Institute Research Communications No. 57, Interleukin 2, F.R. Seiler and H.G. Schwick, eds.; Lymphokines, Vol. 5, Mono- clonal T Cells and Their Products, M. Feldmann and M. Schreier, eds., Academic Press, New York, 1982 16. Ricciardi Castagnoli et al., this volume; Finn et al. (1979) Proc Natl Acad Sci USA 76: 4033 17. Immunol Rev 54 (1981); Nabel et al. (1981) Cell 23: 19 Acknowledgments The Basel Institute for Immunology, founded and supported by F. Hoffmann-La Roche and Co., Ltd., and Springer-Verlag jointly sponsored this workshop. Our sincere thanks are due for their support. The work of B. Badovinac, J. Bron, M. Dekany, R. Rodrigues, J.-P. Rudloff, G. Scialdone, and A. Wyss of the Basel Institute for Immunol- ogy in managing the technical details of this workshop is much appreciated. x We also thank Marga Botsch and Rick Mills of Springer-Verlag for their assistance in the publication of the workshop proceedings. We are grateful to all contributors to this book for meeting the deadline for submission of their manuscripts, which greatly facili- tated rapid publication. The articles appear essentially as sub- mitted by the authors; no editing for style or content has been done. The Organizers XI Lectin-Dependent Cytolytic and Cytolymic T Helper Clones and Hybridomas K. Fischer Lindahl, A.A. Nordin, M.H. Schreier A. Introduction Any cytotoxic T lymphocyte (CTL) will kill P81S or EL4 target cells on addition of a lectin such as phytohemagglutinin (PHA) or concanava- lin A (Con A) (1,2), and lectin-dependent killing has been considered a prerogative of CTL (3). However, lectin-dependent killers could be found under conditions where no specific CTL were induced (4-6), and nearly every T cell c~~ne stimulated by Con A and T cell growth factor (TCGF) caused Cr-release in the presence of PHA (7). It therefore seemed possible that any T cell can become a lectin-depen- dent killer. To find out how cornmon this activity might be, we tested a set of antigen-specific, I-A-restricted T helper cell lines, clones and hybridomas (8-10). B. Materials and Methods Helper Cell Lines and Clones. All T cells were from CS7BL/6 mice. The methods used to establish and clone these cells in vitro have been publishEd together with a functional characterization (8-11). All were I-A -restricted. They had been cultured serum-free for more than a year by continued restimulation with antigen and irradiated CS7BL/6 nu/nu spleen cells or in medium supplemented with TCGF. Before testing, the lines were expanded for at least two weeks in medium with TCGF in the absence of adherent cells and antigen. Hybridomas. All hybridomas originated from two fusions of a chicken egg albumin- (EA) specific clone, EA-14, with BWS147 and are described elsewhere in this volume (12). They were cultured in RPMI 1640 supplemented with S% heat-inactivated fetal calf serum, glutamine, pyruvate, 2-mercaptoethanol, and penicillin-streptomycin. SlCr-Release Assay. A standard method (13) was used with 10 4 SlCr_ labeled target cells and effector-to-target ratios of 10:1, 3:1 and 1:1. Triplicates were set up in round-bottom microtiter plates in 200 ~l of RPMI 1640 with S% heat-inactivated fetal calf serum. PHA-P (Difco) was added to a final concentration of 10 ~g/ml, Con A (Sigma) to S ~g/ml (3 51 and the cultures were incubated for 3.S hours. The experimental Cr-release is expressed as a percentage of the total (measured in the presence of a detergent), and the spontaneous release has been subtracted in the tables. Spleen cells stimulated with Con A for three days or in allogeneic mixed lymphocyte cultures

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