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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):1292–1296. doi: 10.1073/pnas.81.5.1292

Expression of an immunoglobulin heavy chain gene transfected into lymphocytes.

R J Deans, K A Denis, A Taylor, R Wall
PMCID: PMC344819  PMID: 6324184

Abstract

We determined that mouse lymphoid cell lines can be transfected at high efficiencies (10-70%) by a polyoma virus shuttle vector. With this vector, we obtained expression of a cloned mouse alpha heavy chain gene transfected into cell lines representative of all stages in B-lymphocyte development, a T-cell lymphoma line, and 3T3 fibroblasts. Heavy chain gene expression in transfected light chain-producing myeloma cells occurred at levels comparable to those in IgA-secreting myeloma cells. Heavy chains produced in transfected myeloma cells were associated with light chains in membrane-bound IgA. While T-lymphoma cells and fibroblasts were transfected at similar efficiencies to B cells, significantly lower levels of alpha heavy chains were produced. This immunoglobulin gene transfection system provides a powerful approach for defining important regulatory regions in immunoglobulin genes and for identifying lymphoid cell factors involved in immunoglobulin gene expression in B-lymphocyte development.

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Selected References

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  1. Banerji J., Olson L., Schaffner W. A lymphocyte-specific cellular enhancer is located downstream of the joining region in immunoglobulin heavy chain genes. Cell. 1983 Jul;33(3):729–740. doi: 10.1016/0092-8674(83)90015-6. [DOI] [PubMed] [Google Scholar]
  2. Bendig M. M., Thomas T., Folk W. R. Regulatory mutants of polyoma virus defective in DNA replication and the synthesis of early proteins. Cell. 1980 Jun;20(2):401–409. doi: 10.1016/0092-8674(80)90626-1. [DOI] [PubMed] [Google Scholar]
  3. Chen-Kiang S., Wolgemuth D. J., Hsu M. T., Darnell J. E., Jr Transcription and accurate polyadenylation in vitro of RNA from the major late adenovirus 2 transcription unit. Cell. 1982 Mar;28(3):575–584. doi: 10.1016/0092-8674(82)90212-4. [DOI] [PubMed] [Google Scholar]
  4. Chu G., Sharp P. A. SV40 DNA transfection of cells in suspension: analysis of efficiency of transcription and translation of T-antigen. Gene. 1981 Mar;13(2):197–202. doi: 10.1016/0378-1119(81)90008-1. [DOI] [PubMed] [Google Scholar]
  5. Davis M. M., Calame K., Early P. W., Livant D. L., Joho R., Weissman I. L., Hood L. An immunoglobulin heavy-chain gene is formed by at least two recombinational events. Nature. 1980 Feb 21;283(5749):733–739. doi: 10.1038/283733a0. [DOI] [PubMed] [Google Scholar]
  6. Friedmann T., Esty A., LaPorte P., Deininger P. The nucleotide sequence and genome organization of the polyoma early region: extensive nucleotide and amino acid homology with SV40. Cell. 1979 Jul;17(3):715–724. doi: 10.1016/0092-8674(79)90278-2. [DOI] [PubMed] [Google Scholar]
  7. Gariglio P., Llopis R., Oudet P., Chambon P. The template of the isolated native simian virus 40 transcriptional complexes is a minichromosome. J Mol Biol. 1979 Jun 15;131(1):75–105. doi: 10.1016/0022-2836(79)90302-4. [DOI] [PubMed] [Google Scholar]
  8. Gillies S. D., Morrison S. L., Oi V. T., Tonegawa S. A tissue-specific transcription enhancer element is located in the major intron of a rearranged immunoglobulin heavy chain gene. Cell. 1983 Jul;33(3):717–728. doi: 10.1016/0092-8674(83)90014-4. [DOI] [PubMed] [Google Scholar]
  9. Goldsby R. A., Osborne B. A., Simpson E., Herzenberg L. A. Hybrid cell lines with T-cell characteristics. Nature. 1977 Jun 23;267(5613):707–708. doi: 10.1038/267707a0. [DOI] [PubMed] [Google Scholar]
  10. Gronowicz E., Coutinho A., Melchers F. A plaque assay for all cells secreting Ig of a given type or class. Eur J Immunol. 1976 Aug;6(8):588–590. doi: 10.1002/eji.1830060812. [DOI] [PubMed] [Google Scholar]
  11. Hofman F. M., Billing R. J., Parker J. W., Taylor C. R. Cytoplasmic as opposed to surface Ia antigens expressed on human peripheral blood lymphocytes and monocytes. Clin Exp Immunol. 1982 Aug;49(2):355–363. [PMC free article] [PubMed] [Google Scholar]
  12. Honjo T. Immunoglobulin genes. Annu Rev Immunol. 1983;1:499–528. doi: 10.1146/annurev.iy.01.040183.002435. [DOI] [PubMed] [Google Scholar]
  13. Joho R., Nottenburg C., Coffman R. L., Weissman I. L. Immunoglobulin gene rearrangement and expression during lymphocyte development. Curr Top Dev Biol. 1983;18:15–58. doi: 10.1016/s0070-2153(08)60578-5. [DOI] [PubMed] [Google Scholar]
  14. Klinman N. R., Taylor R. B. General methods for the study of cells and serum during the immune response: the response to dinitrophenyl in mice. Clin Exp Immunol. 1969 Apr;4(4):473–487. [PMC free article] [PubMed] [Google Scholar]
  15. Köhler G., Milstein C. Derivation of specific antibody-producing tissue culture and tumor lines by cell fusion. Eur J Immunol. 1976 Jul;6(7):511–519. doi: 10.1002/eji.1830060713. [DOI] [PubMed] [Google Scholar]
  16. Mellon P., Parker V., Gluzman Y., Maniatis T. Identification of DNA sequences required for transcription of the human alpha 1-globin gene in a new SV40 host-vector system. Cell. 1981 Dec;27(2 Pt 1):279–288. doi: 10.1016/0092-8674(81)90411-6. [DOI] [PubMed] [Google Scholar]
  17. Murray N. E., Brammar W. J., Murray K. Lambdoid phages that simplify the recovery of in vitro recombinants. Mol Gen Genet. 1977 Jan 7;150(1):53–61. doi: 10.1007/BF02425325. [DOI] [PubMed] [Google Scholar]
  18. Ochi A., Hawley R. G., Shulman M. J., Hozumi N. Transfer of a cloned immunoglobulin light-chain gene to mutant hybridoma cells restores specific antibody production. Nature. 1983 Mar 24;302(5906):340–342. doi: 10.1038/302340a0. [DOI] [PubMed] [Google Scholar]
  19. Oi V. T., Morrison S. L., Herzenberg L. A., Berg P. Immunoglobulin gene expression in transformed lymphoid cells. Proc Natl Acad Sci U S A. 1983 Feb;80(3):825–829. doi: 10.1073/pnas.80.3.825. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Paige C. J., Kincade P. W., Ralph P. Murine B cell leukemia line with inducible surface immunoglobulin expression. J Immunol. 1978 Aug;121(2):641–647. [PubMed] [Google Scholar]
  21. Picard D., Schaffner W. Correct transcription of a cloned mouse immunoglobulin gene in vivo. Proc Natl Acad Sci U S A. 1983 Jan;80(2):417–421. doi: 10.1073/pnas.80.2.417. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Queen C., Baltimore D. Immunoglobulin gene transcription is activated by downstream sequence elements. Cell. 1983 Jul;33(3):741–748. doi: 10.1016/0092-8674(83)90016-8. [DOI] [PubMed] [Google Scholar]
  23. Rice D., Baltimore D. Regulated expression of an immunoglobulin kappa gene introduced into a mouse lymphoid cell line. Proc Natl Acad Sci U S A. 1982 Dec;79(24):7862–7865. doi: 10.1073/pnas.79.24.7862. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roberts T. M., Kacich R., Ptashne M. A general method for maximizing the expression of a cloned gene. Proc Natl Acad Sci U S A. 1979 Feb;76(2):760–764. doi: 10.1073/pnas.76.2.760. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Rogers J., Early P., Carter C., Calame K., Bond M., Hood L., Wall R. Two mRNAs with different 3' ends encode membrane-bound and secreted forms of immunoglobulin mu chain. Cell. 1980 Jun;20(2):303–312. doi: 10.1016/0092-8674(80)90616-9. [DOI] [PubMed] [Google Scholar]
  26. Rogers J., Wall R. Immunoglobulin heavy chain genes: demethylation accompanies class switching. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7497–7501. doi: 10.1073/pnas.78.12.7497. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Sompayrac L. M., Danna K. J. Efficient infection of monkey cells with DNA of simian virus 40. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7575–7578. doi: 10.1073/pnas.78.12.7575. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Wall R., Kuehl M. Biosynthesis and regulation of immunoglobulins. Annu Rev Immunol. 1983;1:393–422. doi: 10.1146/annurev.iy.01.040183.002141. [DOI] [PubMed] [Google Scholar]
  29. Word C. J., Mushinski J. F., Tucker P. W. The murine immunoglobulin alpha gene expresses multiple transcripts from a unique membrane exon. EMBO J. 1983;2(6):887–898. doi: 10.1002/j.1460-2075.1983.tb01518.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. de Saint Vincent B. R., Delbrück S., Eckhart W., Meinkoth J., Vitto L., Wahl G. The cloning and reintroduction into animal cells of a functional CAD gene, a dominant amplifiable genetic marker. Cell. 1981 Dec;27(2 Pt 1):267–277. doi: 10.1016/0092-8674(81)90410-4. [DOI] [PubMed] [Google Scholar]

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