Abstract
In this study we describe the generation and characterization of interspecies somatic cell hybrids between human activated mature T cells and mouse BW5147 thymoma cells. A preferential segregation of human chromosomes was observed in the hybrids. Phenotypic analysis of two hybrids and their clones demonstrated coexpression of CD4 and CD69 antigens in the same cells. Segregation analysis of an informative family of hybrids followed by molecular and karyotype studies clearly demonstrated that the locus encoding CD69 antigen mapped to human chromosome 12. Although the expression of CD69 antigen is an early event after T-lymphocyte activation and rapidly declines in absence of exogenous stimuli, in the hybrids described in this study the expression was constitutive, similarly to what was previously found in early thymocyte precursors and mature thymocytes. In this respect it was important to note that the behavior of the hybrids in culture strongly suggested a dominant influence of the thymus-derived mouse tumor cell genome in controlling the constitutive expression of human CD69. These hybrids may thus provide a system to study the genetic and molecular mechanisms controlling the expression and function of this activation antigen.
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Accolla, R. S., Scarpellino, L., Carra, G., and Guardiola, J.: Transacting elements operating across species barriers positively regulate expression of major histocompatibility complex class II genes. J Exp Med 162: 1117–1133, 1985
Bjorndahl, J. M., Nakamura, S., Hara, T., Jung, L. K. L., and Fu, S. M.: The 28-kDa 32-kDa activation antigen EA 1. Further characterization and signal requirements for its expression. J Immunol 141: 4094–4100, 1988
Cebrian, M., Yague, E., Rincon, M., Lopez-Botet, M., De Landazuri, M. O., and Sanchez-Madrid, F.: Triggering of T cell proliferation through AIM, an activation inducer molecule expressed on activated human lymphocytes. J Exp Med 168: 1621–1637, 1988
Cebrian, M., Redondo, J. M., Lopez-Rivas, A., Rodriguez-Tarduchy, G., De Landazuri, M. O., and Sanchez-Madrid, F.: Expression and function of AIM, an activation inducer molecule of human lymphocytes, is dependent on the activation of protein kinase C. Eur J Immunol 19: 809–815, 1989
Cosulich, M. E., Rubartelli, A., Risso, A., Cozzolino, F., and Bargellesi, A.: Functional characterization of an antigen involved in an early step of T-cell activation. Proc Natl Acad Sci USA 84: 4205–4209, 1987
Gerosa, F., Tommasi, M., Scardoni, M., Accolla, R. S., Pozzan, T., Libonati, M., Tridente, G., and Carra, G.: Biochemical and functional analysis of the CD69 early activation antigen by two monoclonal antibodies directed to different epitopes. Mol Immunol 28: 159–168, 1991
Hara, T., Jung, L. K. L., Bjorndahl, J. M., and Fu, S. M.: Human T cell activation. III. Rapid induction of a phosphorylated 28 kD/32 kD disulphide-linked early activation antigen (EA 1) by 12-o-tetradecanoyl phorbol-13-acetate, mitogens, and antigens. J Exp Med 164: 1988–2005, 1986
Jung, L. K. L., Haynes, B. F., Nakamura, S., Pahwa, S., and Fu, S. M.: Expression of early activation antigen (CD69) during human thymic development. Clin Exp Immunol 81: 466–474, 1990
Lanier, L. L., Buck, D. W., Rhodes, L., Ding, A., Evans, E., Barney, C., and Phillips, J. H.: Interleukin 2 activation of natural killer cells rapidly induces the expression and phosphorylation of the Leu-23 activation antigen. J Exp Med 167: 1572–1585, 1988
Nakamura, S., Sung, S. J., Bjorndahl, J. M., and Fu, S. M.: Human T cell activation. IV. T cell activation and proliferation via the early activation antigen EA 1. J Exp Med 169: 677–689, 1989
Phillips, J. H., Takeshita, T., Sugumura, K., and Lanier, L. L.: Activation of natural killer cells via the p75 interleukin 2 receptor. J Exp Med 170: 291–296, 1989
Risso, A., Cosulich, M. E., Rubartelli, A., Mazza, M. R., and Bargellesi, A.: MLR3 molecule is an activation antigen shared by human B, T lymphocytes and T cell precursors. Eur J Immunol 19: 323–328, 1989
Risso, A., Smilivich, D., Capra, M. C., Baldissarro, I., Yan, G., Bargellesi, A., and Cosulich, M. E.: CD69 in resting and activated lymphocytes. Its association with a GTP binding protein and biochemical requirements for its expression. J Immunol 146: 4105–4114, 1991
Sambrook, J., Fritsch, E. F., and Maniatis, T.: Molecular Cloning: A Laboratory Manual 2nd edition, Cold Spring Harbor Laboratory, Cold Spring Harbor, 1989
Sanchez-Mateos, P. and Sanchez-Madrid, F.: Structure-function relationship and immunochemical mapping of external and intracellular antigenic sites on the lymphocyte activation inducer molecule, AIM/CD69. Eur J Immunol 21: 2317–2325, 1991
Sartoris, S., Scupoli, M. T., Scarpellino, L., Paiola, F., Jotterand-Bellomo, M., Tridente, G., and Accolla, R. S.: Inducible and constitutive MHC class II gene expression: distinct tissue-specific genetic controls. J Immunol 145: 1960–1967, 1990
Testi, R., Phillips, J. H., and Lanier, L. L.: Constitutive expression of a phosphorylated activation antigen (Leu-23) by CD-3 bright human thymocytes. J Immunol 141: 2557–2563, 1988
Testi, R., Phillips, J. H., and Lanier, L. L.: T cell activation via Leu-23 (CD-69). J Immunol 143: 1123–1128, 1989a
Testi, R., Phillips, J. H., and Lanier, L. L.: Leu23 induction as an early marker of functional CD3/T cell antigen receptor triggering. Requirement for receptor cross-linking, prolonged elevation of intracellular [Ca+ +] and stimulation of protein kinase C. J Immunol 142: 1854–1860, 1989b
Testi, R., Pulcinelli, F., Frati, L., Cazzaniga, P. P., and Santoni, A.: CD69 is expressed on platelets and mediates platelet activation and aggregation. J Exp Med 172: 701–707, 1990
Ulmann, K. S., Northrop, J. P., Verweij, C. L., and Cratbree, G. R.: Transmission of signal from T lymphocyte antigen receptor to the genes responsible for cell proliferation and immune function. Annu Rev Immunol 8: 421–452, 1991
Yokoyama, W. M., Koning, F., Kehn, P. J., Pereira, G. M. B., Stingl, G., Coligan, J. E., and Shevach, E. M.: Characterization of a cell surface-expressed disulfide-linked dimer involved in murine T cell activation. J Immunol 141: 369–376, 1988
Yokoyama, W. M., Maxfield, S. R., and Shevach, E. M.: Very early (VEA) and very late (VLA) activation antigens have distinct functions in T lymphocyte activation. Immunol Rev 109: 153–176, 1989
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Address correspondence and offprint requests to: R. S. Accolla, Istituto di Scienze Immunologiche, Facolta'di Medicina e Chirurgia, Universita'di Verona, Policlinico Borgo Roma, 37100 Verona, Italy.
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Cambiaggi, C., Scupoli, M.T., Cestari, T. et al. Constitutive expression of CD69 in interspecies T-cell hybrids and locus assignment to human chromosome 12. Immunogenetics 36, 117–120 (1992). https://doi.org/10.1007/BF00215288
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DOI: https://doi.org/10.1007/BF00215288