Abstract
The mouse B-cell clone, CH12.LX (Iak, Ly-1+, μ+, δ+), can be induced to differentiate and secrete antibody in an antigen-specific, H-2-restricted manner. Induction requires two signals. One must be provided by the binding of specific antigen to the membrane IgM; the other is delivered by the binding of Ek-specific T-cell hybridomas to the Ek molecules of CH12.LX (Bishop and Haughton 1986). Previous studies demonstrated that Ek-specific monoclonal antibodies (mAbs) could substitute for T cells in delivering the second differentiative signal (Bishop and Haughton 1986). Although CH12.LX cells present Ak to Ak-restricted or alloreactive T-helper cells, neither T cells nor mAbs specific for Ak induce differentiation (Bishop and Haughton 1986). However, since the Akspecifc mAbs tested previously were β-chain-specific and the Ia epitope specificity of the T cells used was unknown, it is possible that the differentiative signal delivered to the CH12.LX class 11 molecule is chain-specific. Here we report the effects of ten additional Iak-specific mAbs upon the differentiation of CH12.LX. In addition, a cl)NA library was prepared from CHI 2.LX cells, clones corresponding to the α and β chains of the Ak molecule were isolated, and their nucleotide sequences were determined. Finally, the Ak and Ek molecules of CH12.LX and H-2k spleen cells were compared by two-dimensional gel electrophoresis to examine possible post-translational differences in the Iak molecules of CH12.LX.
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Arnold, L. W., LoCascio, N. J., Lutz, P. M., Pennell, C. A., Klapper, D., and Haughton, G.: Antigen-induced lymphomagenesis: identification of a murine B cell lymphoma with known antigen specificity. J. Immunol. 131: 2064–2068, 1983
Badley, J. E., Bishop, G. A., St. John, T., and Frelinger, J. A.: A simple, rapid method for the purification of Poly A+ RNA. Biotechniques 6: 114–116, 1988
Bankier, A. T. and Barrell, B. G.: Techniques in Nucleic Acid Chemistry, pp. 1–34, Elsevier, New York, 1983
Benoist, C. O., Mathis, D. J., Kanter, M. R., Williams III, V. E., and McDavitt, H. O.: The murine Ia a chains, Eα and Aα, show a surprising degree of homology. Proc. Natl. Acad. Sci. U.S.A. 80: 534–538, 1983
Benton, W. D. and Davis, R. W.: Screening lambda gt recombinant clones by hybridization to single plaques in situ. Science 196: 180–182, 1977
Bishop, G. A. and Haughton, G.: Induced differentiation of a transformed clone of Ly-1 + B cells by clonal T cells and antigen. Proc. Natl. Acad. Sci. U.S.A. 83: 7410–7414, 1986
Bishop, G. A. and Haughton, G.: Role of the interleukin 2 receptor in differentiation of a clone of Ly-1 + B cells. J. Immunol. 138: 3308–3313, 1987
Cambier, J. C., Newell, M. K., Justement, L. B., McGuire, J. C., Leach, K. L., and Chen, Z. Z.: Ia binding ligands and cAMP stimulate nuclear translocation of PKC in B lymphocytes. Nature 327: 629–632, 1987
Choi, E., McIntyre, K., Germain, R. N., and Seidman, J. G.: Murine I-A β chain polymorphism: nucleotide sequences of three allelic I-A, genes. Science 221: 283–286, 1983
Cunningham, A. J. and Szenberg, A.: Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology 14: 599–601, 1968
Dretzen, G., Bellard, M., Sassone-Corsi, P., and Chambon, P.: A reliable method for the recovery of DNA fragments from agarose and acrylamide gels. Anal. Biochem. 112: 295–298, 1981
Estess, P., Begovich, A. B., Koo, M., Jones, P. P., and McDevitt, H. O.: Sequence analysis and structure-function correlations of murine k, u, s, and f haplotype I-A β cDNA clones. Proc. Natl. Acad. Sci. U.S.A. 83: 3594–3598, 1986
Frelinger, J. A., Niederhuber, J. E., David, C. S., and Shreffler, D. C.: Evidence for the expression of Ia antigens on thymus-derived lymphocytes. J. Exp. Med. 140: 1273–1284, 1974
Frelinger, J. A., Niederhuber, J. E., and Shreffler, D. C.: Inhibition of immune responses in vitro by specific antiserums to Ia antigens. Science 188: 268–270, 1975
Frelinger, J. G., Shigeta, M., and Fathman, C. G.: An analysis of monoclonal T cell and antibody recognition sites on Ia molecules. J. Invest. Dermatol. 85: 30S-40S, 1985
Germain, R. N. and Malissen, B.: Analysis of the expression and function of class II MHC-encoded molecules by DNA-mediated gene transfer. Ann. Rev. Immunol. 4: 281–315, 1986
Gubler, U. and Hoffman, B.: A simple and very efficient method for generating cDNA libraries. Gene 25: 263–269, 1983
Hämmerling, G., Hämmerling, U., and Lemke, H.: Isolation of twelve monoclonal antibodies against Ia and H-2 antigens. Serological characterization and reactivity with B and T lymphocytes. Immunogenetics 8: 433–445, 1979
Henry, C., Chan, E. L., and. Kodlin, D.: Expression and function of I region products on immunocompetent cells. II. I region products in T-B interaction. J. Immunol. 119: 744–748, 1977
Janeway, C. A., Conrad, P. J., Lerner, E. A., Babich, J., Wettstein, P., and Murphy, D. B.: Monoclonal antibodies specific for Ia glycoproteins raised by immunization with activated T cells: possible role of T cellbound Ia antigens as targets of immunoregulatory T cells. J. Immunol. 132: 662–667, 1984
Knapp, M. R., Jones, P. P., Black, S. J., Vitetta, E. S., and Strober, S.: Characterization of a spontaneous murine B cell leukemia (BCL1). I. Cell surface expression of IgM, IgD, Ia, and FcR. J. Immunol. 123: 992–999, 1979
LoCascio, N. J., Arnold, L. W., Corley, R. B., and Haughton, G.: Induced differentiation of a B cell lymphoma with known antigen specificity. J. Mol. Cell. Immunol. 1: 177–187, 1984a
LoCascio, N. J., Haughton, G., and Corley, R. B.: Role of cell surface Ig in B-lymphocyte activation. Proc. Natl. Acad. Sci. U.S.A. 81: 2466–2469, 1984b
McMillan, M., Cecka, J. M., Murphy, D. B., McDevitt, H. O., and Hood, L.: Structure of murine Ia antigens: partial NH2 terminal amino acid sequences of products of the I-E or I-C subregion. Proc. Natl. Acad. Sci. U.S.A. 74: 5135–5139, 1977
McMillan, M., Frelinger, J. A., Jones, P. P., Murphy, D. B., McDevitt, H. O., and Hood, L.: Structure of murine Ia antigens. Two-dimensional electrophoretic analysis and high-pressure liquid chromatography tryptic peptide maps of products of the I-A and I-E subregions and of an associated invariant polypeptide. J. Exp. Med. 153: 936–950, 1981
Mercolino, T. J., Arnold, L. W., and Haughton, G.: Phosphatidyl choline is recognized by a series of Ly1+ murine B cell lymphomas specific for erythrocyte membranes. J. Exp. Med. 163: 155–165, 1986
Monroe, J. G. and Cambier, J. C.: B cell activation. III. B cell plasma membrane depolarization and hyper-Ia antigen expression induced by receptor immunoglobulin cross-linking are coupled. J. Exp. Med. 58: 1589–1599, 1983
Oi, V. T., Jones, P. P., Goding, J. W., Herzenberg, L. A., and Herzenberg, L. A.: Properties of monoclonal antibodies to mouse Ig allotypes, H-2, and Ia antigens. Curr. Top. Microbiol. Immunol. 81: 115–129, 1978
Ozato, K., Mayer, N., and Sachs, D. H.: Hybridoma cell lines secreting monoclonal antibodies to mouse H-2 and Ia antigens. J. Immunol. 124: 533–540, 1980
Palmer, M. J. and Frelinger, J. A.: Widespread transcription of a Qa region gene in adult mice. J. Exp. Med. 166: 95–108, 1987
Pierres, M., Devaux, C., Dosseto, M., and Marchetto, S.: Clonal analysis of B- and T-cell responses to Ia antigens. I. Topology of epitope regions of I-Ak and I-Ek molecules analyzed with 35 monoclonal alloantibodies. Immunogenetics 14: 481–495, 1981
Rigby, P. W. J., Dieckman, M., Rhodes, C., and Berg, P.: Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J. Mol. Biol. 113: 237–251, 1977
Sanger, F., Coulson, A. R., Barrell, B. G., Smith, A. J. H., and Roe, B. A.: Cloning in a single-stranded bacteriophage as an aid to rapid DNA sequencing. J. Mol. Biol. 143: 161–178, 1980
Schwartz, R. H.: T-lymphocyte recognition of antigen in association with gene products of the major histocompatibility complex. Annu. Rev. Immunol. 3: 237–261, 1985
Takiguchi, M. and Frelinger, J. A.: Induction of antigen presentation ability in purified cultures of astroglia by interferon gamma. J. Mol. Cell. Immunol. 2: 269–280, 1986
Zagursky, R. J., Baumeister, K., Lomax, N., and Berman, M. L.: Rapid and easy sequencing of large linear double-stranded DNA and supercoiled plasmid DNA. Gene Anal. Techn. 2: 89–94, 1985
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Bishop, G.A., McMillan, M.S., Haughton, G. et al. Signaling to a B-cell clone by Ek, but not Ak, does not reflect alteration of A k genes. Immunogenetics 28, 184–192 (1988). https://doi.org/10.1007/BF00375858
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DOI: https://doi.org/10.1007/BF00375858