Abstract
A panel of twleve mutant A kβ expressing cell lines has been produced from a mutagenized B cell hybridoma using a protocol of negative and positive immunoselection. Each of the serologically-defined epitopes on the A kβ polypeptide has been altered in one or more of the mutant cell lines. Immunoselection techniques have been used to select also for secondary mutations in some cell lines, resulting either in reversion to the wild-type or near wild-type phenotype or in the alteration of additional A kβ serologic epitopes. Characterization of the ability of the mutant cell lines to stimulate antigen and alloantigen-reactive T cell hybridomas has shown that they display distinct functional phenotypes ranging from nearly wild-type to almost completely defective. DNA sequence analysis of the A kβ gene cloned from each mutant cell line has revealed the structural basis for the mutant serologic and functional phenotypes. To date, each β chain mutant has been found to have a single amino acid substitution (resulting from a single nucleotide substitution) in the β1 domain. The substitutions are clustered in or near the third allelicly-defined variable region of the Aβ polypeptide (residues 61–70). These results indicate that this region of the Aβ polypeptide comprises multiple serologically defined epitopes. Furthermore, specific residues within this region appear to be important for effective T cell activation.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Beck, B. N., Glimcher, L. H., Nilson, A. E., Pierres, M., and McKean, D. J., 1984, The structure-function relationship of I-A molecules: Correlation of serologic and functional phenotypes of four I-Ak mutant cell lines. J. Immuno1., 133: 3176.
Beck, B. N., Pease, L. R., Bell, M. P., Buerstedde, J.-M., Nilson, A. E., Schlauder, G. G., and McKean, D. J., 1987, DNA sequence analysis of I-Aß mutants reveals serologically immunodominant region. J. Exp. Med., in press.
Benoist, C. O., Mathis, D. J., Kanter, M. R., Williams, V. E., II, McDevitt, H. O., 1983, Regions of allelic hypervariability in the murina a immune response gene. Cell, 34: 169.
Brown, M. A., Glimcher, L. H., Nielsen, E. A., Paul, W. E., and Germain, R. N., 1986, T-cell recognition of Ia molecules selectively altered by a single amino acid substitution. Science, 231: 255.
Choi, E., McIntyre, K., Germain, R. N., and Seidman, J. G., 1983, Murine I-Aß chain polymorphism: Nucleotide sequences of three allelic I-Aß genes. Science, 221: 283.
Cohn, L. E., Glimcher, L. H., Waldmann, R. A., Smith, J. A., Ben-Nun, A., Seidman, J. G., and Choi E., 1986, Identification of functional regions on the I-Ab molecule by site-directed mutagenesis. Proc. Natl. Acad. Sci. USA, 83: 747.
Estess, P., Begovich, A. B., Koo, M., Jones, P. P., and McDevitt, H. O., 1986, Sequence analysis and structure-function correlations of murine q, k, u, s, and f haplotype I-A8 cDNA clones. Proc. Natl. Acad. Sci. USA, 83: 3594.
Germain, R. N., Ashwell, J. D., Lechler, R. I., Margulies, D. H., Nickerson, K. M., Suzuki, G., and Tou, J. Y. L., 1985, “Exon-shuffling” maps control of antibody and T-cell-recognition sites to the NH2-terminal domain of the class II major histocompatibility polypeptide A8. Proc. Natl. Acad. Sci. USA, 82: 2940.
Glimcher, L. H., Hamano, T., Asofsky, R., Sachs, D. H., Pierres, M., Samelson, L. E., Sharrow, S. O., and Paul, W. E., 1983a, I-A mutant functional antigen-presenting cell lines. J. Immunol., 130: 2287.
Glimcher, L. H., Sharrow, S. O., and Paul, W. E., 1983b, Serologic and functional characterization of a panel of antigen-presenting cell lines expressing mutant I-A class II molecules. J. Exp. Med., 158: 1523.
Harmon, R. C., Stein, N., and Frelinger, J. A., 1983, Monoclonal antibodies reactive with H-2 determinants. Immunogenetics, 18: 541.
Landais, D., Matthes, H., Benoist, C., and Mathis, D., 1985, A molecular basis for the Ia.2 and Ia.19 antigenic determinants. Proc. Natl. Acad. Sci. USA, 83, 747.
McIntyre, K. R., and Seidman, J. G., 1984, Nucleotide sequence of mutant I-Abm12 gene is evidence for genetic exchange between mouse immune response genes. Nature, 308, 551.
Oi, V. T., Jones, P. P., Coding, J. W., Herzenberg, L. A., and Herzenberg, L. A., 1978, Properties of monoclonal antibodies to mouse Ig allotypes, H-2, and Ia antigens. Curr. Top. Microbiol. Immunol., 81: 115.
Pierres, M., Devaux, C., Dosseto, N., and Marchetto, S., 1981, Clonal analysis of B- and T-cell responses to la antigens. I. Topology of epitope regions on I-Ak and I-Ek molecules analyzed with 35 monoclonal antibodies. Immunogenetics, 14: 481.
Robinson, R. R., Germain, R. N., McKean, D. J., Mescher, M., and Seidman, J. G., 1983, Extensive polymorphism surrounding the murine Ia Aß chain gene. J. Immunol., 131: 2025.
Sanger, F., Nicklen, S., Coulson, A. R., 1977, DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. USA, 74: 5463.
Schlauder, G. G., Bell, M. P., Beck, B. N., Nilson, A., and McKean, D. J., 1985, The structure-function relationship of I-A molecules: A biochemical analysis of I-A polypeptides from mutant antigen-presenting cells and evidence of preferential association of allelic forms. J. Immunol., 135: 1945.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1987 Springer Science+Business Media New York
About this chapter
Cite this chapter
Beck, B.N. et al. (1987). Structure-Function Analysis of Serologically Selected Cell Lines Expressing Mutant A kβ Polypeptides. In: David, C.S. (eds) H-2 Antigens. NATO ASI Series, vol 144. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-0764-9_25
Download citation
DOI: https://doi.org/10.1007/978-1-4757-0764-9_25
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4757-0766-3
Online ISBN: 978-1-4757-0764-9
eBook Packages: Springer Book Archive