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. 1983;2(9):1591–1595. doi: 10.1002/j.1460-2075.1983.tb01629.x

Somatic variants of the level of expression of a cell surface antigen.

O R Burrone 1, F Calabi 1, R F Kefford 1, C Milstein 1
PMCID: PMC555328  PMID: 11892817

Abstract

Somatic variants with constitutive changes in the expression of the cortical thymocyte differentiation antigen HTA 1 were derived from the T-cell leukemia line Molt 4 using monoclonal antibody NA1/34 and the fluorescent activated cell sorter. Cells with the highest and lowest fluorescence were sorted and expanded. After several cycles, high expressor variants, with 10- to 12-fold increased surface HTA 1 and low expressors, with a level of one third relative to the wild-type, were obtained. The stability of the high expressors was improved by cloning. In spite of the large differences in the level of HTA 1 between the various mutants and the wild-type, the expression of HLA-A,B,C or its increase following interferon-alpha stimulation, remained unchanged. Although in HTA 1 there was only a trace of beta2-microglobulin (beta2m) detected by lactoperoxidase labelling of the high expressor variants, significant levels of both beta2m and HTA 1 light chain beta(t) were observed in gels stained with Coomassie blue. The physiological association of beta(t) with HTA 1 was confirmed by the substantial increase of beta(t) which parallels the increase of HTA 1.

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Selected References

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

  1. Barnstable C. J., Bodmer W. F., Brown G., Galfre G., Milstein C., Williams A. F., Ziegler A. Production of monoclonal antibodies to group A erythrocytes, HLA and other human cell surface antigens-new tools for genetic analysis. Cell. 1978 May;14(1):9–20. doi: 10.1016/0092-8674(78)90296-9. [DOI] [PubMed] [Google Scholar]
  2. Bradstock K. F., Janossy G., Pizzolo G., Hoffbrand A. V., McMichael A., Pilch J. R., Milstein C., Beverley P., Bollum F. J. Subpopulations of normal and leukemic human thymocytes: an analysis with the use of monoclonal antibodies. J Natl Cancer Inst. 1980 Jul;65(1):33–42. [PubMed] [Google Scholar]
  3. Brodsky F. M., Bodmer W. F., Parham P. Characterization of a monoclonal anti-beta 2-microglobulin antibody and its use in the genetic and biochemical analysis of major histocompatibility antigens. Eur J Immunol. 1979 Jul;9(7):536–545. doi: 10.1002/eji.1830090709. [DOI] [PubMed] [Google Scholar]
  4. Burrone O. R., Milstein C. Control of HLA-A,B,C synthesis and expression in interferon-treated cells. EMBO J. 1982;1(3):345–349. doi: 10.1002/j.1460-2075.1982.tb01172.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Burrone O. R., Milstein C. The effect of interferon on the expression of human cell-surface antigens. Philos Trans R Soc Lond B Biol Sci. 1982 Sep 24;299(1094):133–135. doi: 10.1098/rstb.1982.0116. [DOI] [PubMed] [Google Scholar]
  6. Galfrè G., Milstein C. Preparation of monoclonal antibodies: strategies and procedures. Methods Enzymol. 1981;73(Pt B):3–46. doi: 10.1016/0076-6879(81)73054-4. [DOI] [PubMed] [Google Scholar]
  7. HUNTER M. J., COMMERFORD S. L. Pressure homogenization of mammalian tissues. Biochim Biophys Acta. 1961 Mar 4;47:580–586. doi: 10.1016/0006-3002(61)90553-4. [DOI] [PubMed] [Google Scholar]
  8. Holtkamp B., Cramer M., Lemke H., Rajewsky K. Isolation of a cloned cell line expressing variant H-2Kk using fluorescence-activated cell sorting. Nature. 1981 Jan 1;289(5793):66–68. doi: 10.1038/289066a0. [DOI] [PubMed] [Google Scholar]
  9. Kamarck M. E., Barbosa J. A., Ruddle F. H. Somatic cell genetic analysis of HLA-A, B, C and human beta 2-microglobulin expression. Somatic Cell Genet. 1982 May;8(3):385–402. doi: 10.1007/BF01538895. [DOI] [PubMed] [Google Scholar]
  10. Knowles R. W., Bodmer W. F. A monoclonal antibody recognizing a human thymus leukemia-like antigen associated with beta 2-microglobulin. Eur J Immunol. 1982 Aug;12(8):676–681. doi: 10.1002/eji.1830120810. [DOI] [PubMed] [Google Scholar]
  11. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  12. McMichael A. J., Pilch J. R., Galfré G., Mason D. Y., Fabre J. W., Milstein C. A human thymocyte antigen defined by a hybrid myeloma monoclonal antibody. Eur J Immunol. 1979 Mar;9(3):205–210. doi: 10.1002/eji.1830090307. [DOI] [PubMed] [Google Scholar]
  13. Minowada J., Onuma T., Moore G. E. Rosette-forming human lymphoid cell lines. I. Establishment and evidence for origin of thymus-derived lymphocytes. J Natl Cancer Inst. 1972 Sep;49(3):891–895. [PubMed] [Google Scholar]
  14. Neuberger M. S., Rajewsky K. Switch from hapten-specific immunoglobulin M to immunoglobulin D secretion in a hybrid mouse cell line. Proc Natl Acad Sci U S A. 1981 Feb;78(2):1138–1142. doi: 10.1073/pnas.78.2.1138. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. O'Farrell P. H. High resolution two-dimensional electrophoresis of proteins. J Biol Chem. 1975 May 25;250(10):4007–4021. [PMC free article] [PubMed] [Google Scholar]
  16. Schimke R. T., Alt F. W., Kellems R. E., Kaufman R. J., Bertino J. R. Amplification of dihydrofolate reductase genes in methotrexate-resistant cultured mouse cells. Cold Spring Harb Symp Quant Biol. 1978;42(Pt 2):649–657. doi: 10.1101/sqb.1978.042.01.067. [DOI] [PubMed] [Google Scholar]
  17. Secher D. S., Burke D. C. A monoclonal antibody for large-scale purification of human leukocyte interferon. Nature. 1980 Jun 12;285(5765):446–450. doi: 10.1038/285446a0. [DOI] [PubMed] [Google Scholar]
  18. Ziegler A., Harrison S. C., Leberman R. The minor proteins in tomato bushy stunt and turnip crinkle viruses. Virology. 1974 Jun;59(2):509–515. doi: 10.1016/0042-6822(74)90460-7. [DOI] [PubMed] [Google Scholar]
  19. Ziegler A., Milstein C. A small polypeptide different from beta2-microglobin associated with a human cell surface antigen. Nature. 1979 May 17;279(5710):243–244. doi: 10.1038/279243a0. [DOI] [PubMed] [Google Scholar]

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