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. 1976 Aug 1;144(2):476–493. doi: 10.1084/jem.144.2.476

The Ly-3 antigens on mouse thymocytes: immune precipitation and molecular weight characterization

PMCID: PMC2190382  PMID: 822115

Abstract

Specific anti-Ly sera were employed to precipitate Ly antigens from Nonidet P-40 extracts of mouse thymocytes labeled with 125I using lactoperoxidase and with NaB3H4 using galactose oxidase. Thymocytes from mice of the congenic strains C57BL/6J (Ly-2.2, Ly-3.2 positive), C57BL/6Ly-2a, Ly-3a (Ly-2.1, Ly-3.1 positive) and C57BL/6-Ly-2a (Ly- 2.1, Ly-3.2-positive) were used as sources of labeled antigens and as immune adsorbants to permit evaluation of the specificity of each anti- Ly serum employed. Results of sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing conditions are consistent with the Ly-3.1 antigen containing a glycoprotein subunit with an apparent mol wt of 35,000 daltons. Specific precipitates obtained using anti-Ly-2.1 serum yielded SDS-PAGE profiles identical to that obtained with anti-Ly-3.1 serum, suggesting that the Ly-2 and Ly-3 antigens have the same molecular weight distribution. The relationships of these results to the observed close genetic and topological linkage of Ly-2 and Ly-3 and to the genetic linkage of these loci with the IB-peptide marker, a mouse Bk-region polymorphism, are discussed.

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Selected References

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

  1. Armerding D., Katz D. H. Activation of T and B lymphocytes in vitro. II. Biological and biochemical properties of an allogeneic effect factor (AEF) active in triggering specific B lymphocytes. J Exp Med. 1974 Jul 1;140(1):19–37. doi: 10.1084/jem.140.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Armerding D., Sachs D. H., Katz D. H. Activation of T and B lymphocytes in vitro. III. Presence of Ia determinants on allogeneic effect factor. J Exp Med. 1974 Dec 1;140(6):1717–1722. doi: 10.1084/jem.140.6.1717. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Boyse E. A., Itakura K., Stockert E., Iritani C. A., Miura M. Ly-C: a third locus specifying alloantigens expressed only on thymocytes and lymphocytes. Transplantation. 1971 Mar;11(3):351–353. [PubMed] [Google Scholar]
  4. Boyse E. A., Miyazawa M., Aoki T., Old L. J. Ly-A and Ly-B: two systems of lymphocyte isoantigens in the mouse. Proc R Soc Lond B Biol Sci. 1968 Jun 11;170(1019):175–193. doi: 10.1098/rspb.1968.0032. [DOI] [PubMed] [Google Scholar]
  5. Boyse E. A., Old L. J., Stockert E. An approach to the mapping of antigens on the cell surface. Proc Natl Acad Sci U S A. 1968 Jul;60(3):886–893. doi: 10.1073/pnas.60.3.886. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cantor H., Boyse E. A. Functional subclasses of T lymphocytes bearing different Ly antigens. II. Cooperation between subclasses of Ly+ cells in the generation of killer activity. J Exp Med. 1975 Jun 1;141(6):1390–1399. doi: 10.1084/jem.141.6.1390. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Cantor H., Boyse E. A. Functional subclasses of T-lymphocytes bearing different Ly antigens. I. The generation of functionally distinct T-cell subclasses is a differentiative process independent of antigen. J Exp Med. 1975 Jun 1;141(6):1376–1389. doi: 10.1084/jem.141.6.1376. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Cullen S. E., David C. S., Shreffler D. C., Nathenson S. G. Membrane molecules determined by the H-2 associated immune response region: isolation and some properties. Proc Natl Acad Sci U S A. 1974 Mar;71(3):648–652. doi: 10.1073/pnas.71.3.648. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Edelman G. M., Gottlieb P. D. A genetic marker in the variable region of light chains of mouse immunoglobulins. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1192–1199. doi: 10.1073/pnas.67.3.1192. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Gahmberg C. G., Hakomori S. I. Altered growth behavior of malignant cells associated with changes in externally labeled glycoprotein and glycolipid. Proc Natl Acad Sci U S A. 1973 Dec;70(12):3329–3333. doi: 10.1073/pnas.70.12.3329. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Gall W. E., Edelman G. M. The covalent structure of a human gamma G-immunoglobulin. X. Intrachain disulfide bonds. Biochemistry. 1970 Aug 4;9(16):3188–3196. doi: 10.1021/bi00818a011. [DOI] [PubMed] [Google Scholar]
  12. Gottlieb P. D. Genetic and structural studies of a V-region marker in mouse immunoglobulin light chains.?1780. Contemp Top Mol Immunol. 1976;5:185–216. doi: 10.1007/978-1-4684-8142-6_7. [DOI] [PubMed] [Google Scholar]
  13. Gottlieb P. D. Genetic correlation of a mouse light chain variable region marker with a thymocyte surface antigen. J Exp Med. 1974 Nov 1;140(5):1432–1437. doi: 10.1084/jem.140.5.1432. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Haustein D., Marchalonis J. J., Harris A. W. Immunoglobulin of T lymphoma cells. Biosynthesis, surface representation, and partial characterization. Biochemistry. 1975 May 6;14(9):1826–1834. doi: 10.1021/bi00680a004. [DOI] [PubMed] [Google Scholar]
  15. Henning R., Milner R. J., Reske K., Cunningham B. A., Edelman G. M. Subunit structure, cell surface orientation, and partial amino-acid sequences of murine histocompatibility antigens. Proc Natl Acad Sci U S A. 1976 Jan;73(1):118–122. doi: 10.1073/pnas.73.1.118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Hill R. L., Delaney R., Fellows R. E., Lebovitz H. E. The evolutionary origins of the immunoglobulins. Proc Natl Acad Sci U S A. 1966 Dec;56(6):1762–1769. doi: 10.1073/pnas.56.6.1762. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Hynes R. O. Alteration of cell-surface proteins by viral transformation and by proteolysis. Proc Natl Acad Sci U S A. 1973 Nov;70(11):3170–3174. doi: 10.1073/pnas.70.11.3170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Itakura K., Hutton J. J., Boyse E. A., Old L. J. Genetic linkage relationships of loci specifying differentiation alloantigens in the mouse. Transplantation. 1972 Mar;13(3):239–243. doi: 10.1097/00007890-197203000-00007. [DOI] [PubMed] [Google Scholar]
  19. Itakura K., Hutton J. J., Boyse E. A., Old L. J. Linkage groups of the theta and Ly-A loci. Nat New Biol. 1971 Mar 24;230(12):126–126. doi: 10.1038/newbio230126a0. [DOI] [PubMed] [Google Scholar]
  20. Nathenson S. G., Cullen S. E. Biochemical properties and immunochemical-genetic relationships of mouse H-2 alloantigens. Biochim Biophys Acta. 1974 Apr 8;344(1):1–25. doi: 10.1016/0304-4157(74)90006-9. [DOI] [PubMed] [Google Scholar]
  21. Rombauts W. A., Schroeder W. A., Morrison M. Bovine lactoperoxidase. Partial characterization of the further purified protein. Biochemistry. 1967 Oct;6(10):2965–2977. doi: 10.1021/bi00862a002. [DOI] [PubMed] [Google Scholar]
  22. Segrest J. P., Jackson R. L., Andrews E. P., Marchesi V. T. Human erythrocyte membrane glycoprotein: a re-evaluation of the molecular weight as determined by SDS polyacrylamide gel electrophoresis. Biochem Biophys Res Commun. 1971 Jul 16;44(2):390–395. doi: 10.1016/0006-291x(71)90612-7. [DOI] [PubMed] [Google Scholar]
  23. Shreffler D. C., David C. S. The H-2 major histocompatibility complex and the I immune response region: genetic variation, function, and organization. Adv Immunol. 1975;20:125–195. doi: 10.1016/s0065-2776(08)60208-4. [DOI] [PubMed] [Google Scholar]
  24. Sonoda S., Schlamowitz M. Studies of 125I trace labeling of immunoglobulin G by chloramine-T. Immunochemistry. 1970 Nov;7(11):885–898. doi: 10.1016/0019-2791(70)90051-0. [DOI] [PubMed] [Google Scholar]
  25. Takemori T., Tada T. Properties of antigen-specific suppressive T-cell factor in the regulation of antibody response of the mouse. I. In vivo activity and immunochemical characterization. J Exp Med. 1975 Nov 1;142(5):1241–1253. doi: 10.1084/jem.142.5.1241. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Vitetta E. S., Artzt K., Bennett D., Boyse E. A., Jacob F. Structural similarities between a product of the T/t-locus isolated from sperm and teratoma cells, and H-2 antigens isolated from splenocytes. Proc Natl Acad Sci U S A. 1975 Aug;72(8):3215–3219. doi: 10.1073/pnas.72.8.3215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Vitetta E. S., Bianco C., Nussenzweig V., Uhr J. W. Cell surface immunoglobulin. IV. Distribution among thymocytes, bone mrrow cells, and their derived populations. J Exp Med. 1972 Jul 1;136(1):81–93. doi: 10.1084/jem.136.1.81. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Vitetta E. S., Uhr J. W., Boyse E. A. Association of a beta2-microglobulin-like subunit with H-2 and TL alloantigens on murine thymocytes. J Immunol. 1975 Jan;114(1 Pt 1):252–254. [PubMed] [Google Scholar]
  29. Watkins W. M. Blood-group substances. Science. 1966 Apr 8;152(3719):172–181. doi: 10.1126/science.152.3719.172. [DOI] [PubMed] [Google Scholar]
  30. Weber K., Osborn M. The reliability of molecular weight determinations by dodecyl sulfate-polyacrylamide gel electrophoresis. J Biol Chem. 1969 Aug 25;244(16):4406–4412. [PubMed] [Google Scholar]

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