Skip to main content
NCBI home page
Proceedings of the National Academy of Sciences of the United States of America logoLink to Proceedings of the National Academy of Sciences of the United States of America
. 1984 Mar;81(5):1524–1528. doi: 10.1073/pnas.81.5.1524

Messenger RNA for an antigen-specific binding molecule from an antigen-specific T-cell hybrid.

K D Beaman, N H Ruddle, A L Bothwell, R E Cone
PMCID: PMC344869  PMID: 6200878

Abstract

The T-cell hybridoma 51H7D specifically binds the hapten azobenzenearsonate (ABA). An antiserum (anti-PCIF) specific for antigenic determinants shared by antigen-binding molecules of T cells (TABM) was used to precipitate polyribosomes containing mRNA for TABM from this T-cell hybridoma. The resultant mRNA was translated in vitro. The translated product (TrP51H7D) bound specifically ABA and was bound by both anti-PCIF and anti-T-cell suppressor factor. The parent lymphoma BW5147 yielded a similar translated product with the same antiserum used to isolate specific mRNA containing polysomes. This product (TrPBW) was bound by the antiserum used but did not bind ABA. The specific translated protein from both cells had a pI of approximately equal to 5.0, and apparent molecular weight of 71,000 (reduced) or 145,000 (nonreduced). Both protein products, when treated with guanidine to break down all noncovalent bonds, revealed an elemental peptide of Mr 23,500. The cDNA made from the isolated mRNA had 600-900 bases. mRNA of this size is expected for a protein of Mr 25,000. Our data indicate that a TABM specific for ABA is composed of peptides of Mr 23,500.

Full text

PDF
1524

Images in this article

1526Image
on p.1526
1526Image
on p.1526
1527Image
on p.1527

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Allison J. P., McIntyre B. W., Bloch D. Tumor-specific antigen of murine T-lymphoma defined with monoclonal antibody. J Immunol. 1982 Nov;129(5):2293–2300. [PubMed] [Google Scholar]
  2. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a gamma 2a variable region. Cell. 1981 Jun;24(3):625–637. doi: 10.1016/0092-8674(81)90089-1. [DOI] [PubMed] [Google Scholar]
  3. Clark A. F., Capra J. D. Ubiquitous nonimmunoglobulin p-azobenzenearsonate-binding molecules from lymphoid cells. J Exp Med. 1982 Feb 1;155(2):611–616. doi: 10.1084/jem.155.2.611. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Cone R. E. Molecular basis for T lymphocyte recognition of antigens. Prog Allergy. 1981;29:182–221. [PubMed] [Google Scholar]
  5. Cone R. E., Rosenstein R. W., Janeway C. A., Iverson G. M., Murray J. H., Cantor H., Fresno M., Mattingly J. A., Cramer M., Krawinkel U. Affinity-purified antigen-specific products produced by T cells share epitopes recognized by heterologous antisera raised against several different antigen-specific products from T cells. Cell Immunol. 1983 Dec;82(2):232–245. doi: 10.1016/0008-8749(83)90158-2. [DOI] [PubMed] [Google Scholar]
  6. Cone R. E., Rosenstein R. W., Murray J. H., Iverson G. M., Ptak W., Gershon R. K. Characterization of T-cell surface proteins bound by heterologous antisera to antigen-specific T-cell products. Proc Natl Acad Sci U S A. 1981 Oct;78(10):6411–6415. doi: 10.1073/pnas.78.10.6411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cone R. E., Rosenstein R. W., Tite J., Ptak W., Gershon R. K. Heterologous antisera to Lyt-1+, 2- -derived antigen-binding factor detect a subfactor of an antigen-specific suppressor factor and cell surface proteins on Lyt-1+, 2- and Lyt-1-, 2+ T cells. J Immunol. 1983 May;130(5):2083–2087. [PubMed] [Google Scholar]
  8. Goodman J. W., Lewis G. K., Primi D., Hornbeck P., Ruddle N. H. Antigen-specific molecules from murine T lymphocytes and T cell hybridomas. Mol Immunol. 1980 Jul;17(7):933–945. doi: 10.1016/0161-5890(80)90042-5. [DOI] [PubMed] [Google Scholar]
  9. Haskins K., Kubo R., White J., Pigeon M., Kappler J., Marrack P. The major histocompatibility complex-restricted antigen receptor on T cells. I. Isolation with a monoclonal antibody. J Exp Med. 1983 Apr 1;157(4):1149–1169. doi: 10.1084/jem.157.4.1149. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Hunt J. C., Vasta G. R., Marchalonis J. J. Antigenic and structural characteristics of an immunoglobulin-related peptide purified from marmoset T-lymphoma cells. Cell Immunol. 1983 Jan;75(1):22–42. doi: 10.1016/0008-8749(83)90302-7. [DOI] [PubMed] [Google Scholar]
  11. Isakson P. C., Honegger J. L., Kinsky S. C. Preparation of stable erythrocyte target cells suitable for detection of the antibody response to the haptens azobenzenearsonate, azophenyltrimethylammonium and azophenylphosphorylcholine by plaque-forming cell assay. J Immunol Methods. 1979;25(1):89–96. doi: 10.1016/0022-1759(79)90169-8. [DOI] [PubMed] [Google Scholar]
  12. Lheureux M., Lheureux M., Vervoort T., Van Meirvenne N., Steinert M. Immunological purification and partial characterization of variant-specific surface antigen messenger RNA of Trypanosoma brucei brucei. Nucleic Acids Res. 1979 Oct 10;7(3):595–609. doi: 10.1093/nar/7.3.595. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Palmiter R. D., Christensen A. K., Schimke R. T. Organization of polysomes from pre-existing ribosomes in chick oviduct by a secondary administration of either estradiol or progesterone. J Biol Chem. 1970 Feb 25;245(4):833–845. [PubMed] [Google Scholar]
  14. Ptak W., Gershon R. K., Rosenstein R. W., Murray J. H., Cone R. E. Purification and characterization of TNP-specific immunoregulatory molecules produced by T cells sensitized by picrylchloride (PC1F). J Immunol. 1983 Dec;131(6):2859–2863. [PubMed] [Google Scholar]
  15. Ruddle N. H., Beezley B., Lewis G. K., Goodman J. W. Antigen specific T cell hybrids--II. T cell hybrids which bind azobenzenearsonate. Mol Immunol. 1980 Jul;17(7):925–931. doi: 10.1016/0161-5890(80)90041-3. [DOI] [PubMed] [Google Scholar]
  16. Takeda A., Waldron J. A., Jr, Ruddle N. H., Cone R. E. Analysis of normal and neoplastic lymphocyte surface-labeled proteins by two-dimensional polyacrylamide gel electrophoresis. Exp Cell Res. 1983 Oct;148(1):83–93. doi: 10.1016/0014-4827(83)90189-1. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES