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. 1994 Feb 15;91(4):1323–1327. doi: 10.1073/pnas.91.4.1323

Switch recombination in normal IgA1+ B lymphocytes.

J Irsch 1, S Irlenbusch 1, J Radl 1, P D Burrows 1, M D Cooper 1, A H Radbruch 1
PMCID: PMC43150  PMID: 8108410

Abstract

Most B lymphocytes in normal individuals express two classes of cell-surface immunoglobulins, IgM and IgD. The specificity of the two antigen receptors is identical since they are produced by transcription and differential splicing of the same variable region gene segment to the heavy-chain constant region gene segments for both mu and delta heavy chains. B lymphocytes expressing other immunoglobulin isotypes, IgG, IgA, or IgE, are rare and not well characterized. Particularly controversial is the molecular mechanism of their isotype switch. Here we use high-gradient magnetic cell sorting and fluorescence-activated cell sorting to purify surface IgA1-bearing B lymphocytes from human blood for cellular and molecular analysis. These cells express no immunoglobulin class other than IgA1 and are a relatively uniform population with regard to expression of other cell-surface molecules. They are resting cells in terms of cell cycle and activation marker analysis. The molecular basis for class switching in the IgA1+ cells is not differential transcription or splicing. Rather, switch recombination involving deletion of DNA has occurred on both immunoglobulin heavy-chain gene loci, including the allelically excluded one, and appears to have been directed to IgA1 under normal physiological conditions.

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Selected References

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

  1. Blattner F. R., Tucker P. W. The molecular biology of immunoglobulin D. Nature. 1984 Feb 2;307(5950):417–422. doi: 10.1038/307417a0. [DOI] [PubMed] [Google Scholar]
  2. Borzillo G. V., Cooper M. D., Kubagawa H., Landay A., Burrows P. D. Isotype switching in human B lymphocyte malignancies occurs by DNA deletion: evidence for nonspecific switch recombination. J Immunol. 1987 Aug 15;139(4):1326–1335. [PubMed] [Google Scholar]
  3. Chan M. A., Benedict S. H., Dosch H. M., Hui M. F., Stein L. D. Expression of IgE from a nonrearranged epsilon locus in cloned B-lymphoblastoid cells that also express IgM. J Immunol. 1990 May 1;144(9):3563–3568. [PubMed] [Google Scholar]
  4. Chen Y. W., Word C. J., Jones S., Uhr J. W., Tucker P. W., Vitetta E. S. Double isotype production by a neoplastic B cell line. I. Cellular and biochemical characterization of a variant of BCL1 that expresses and secretes both IgM and IgG1. J Exp Med. 1986 Aug 1;164(2):548–561. doi: 10.1084/jem.164.2.548. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Chen Y. W., Word C., Dev V., Uhr J. W., Vitetta E. S., Tucker P. W. Double isotype production by a neoplastic B cell line. II. Allelically excluded production of mu and gamma 1 heavy chains without CH gene rearrangement. J Exp Med. 1986 Aug 1;164(2):562–579. doi: 10.1084/jem.164.2.562. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Crissman H. A., Hofland M. H., Stevenson A. P., Wilder M. E., Tobey R. A. Supravital cell staining with Hoechst 33342 and DiOC5(3). Methods Cell Biol. 1990;33:89–95. doi: 10.1016/s0091-679x(08)60514-2. [DOI] [PubMed] [Google Scholar]
  7. Esser C., Radbruch A. Immunoglobulin class switching: molecular and cellular analysis. Annu Rev Immunol. 1990;8:717–735. doi: 10.1146/annurev.iy.08.040190.003441. [DOI] [PubMed] [Google Scholar]
  8. Hulett H. R., Bonner W. A., Sweet R. G., Herzenberg L. A. Development and application of a rapid cell sorter. Clin Chem. 1973 Aug;19(8):813–816. [PubMed] [Google Scholar]
  9. Irsch J., Hendriks R., Tesch H., Schuurman R., Radbruch A. Evidence for a human IgG1 class switch program. Eur J Immunol. 1993 Feb;23(2):481–486. doi: 10.1002/eji.1830230227. [DOI] [PubMed] [Google Scholar]
  10. Islam K. B., Nilsson L., Sideras P., Hammarström L., Smith C. I. TGF-beta 1 induces germ-line transcripts of both IgA subclasses in human B lymphocytes. Int Immunol. 1991 Nov;3(11):1099–1106. doi: 10.1093/intimm/3.11.1099. [DOI] [PubMed] [Google Scholar]
  11. Kato K., Radbruch A. Isolation and characterization of CD34+ hematopoietic stem cells from human peripheral blood by high-gradient magnetic cell sorting. Cytometry. 1993;14(4):384–392. doi: 10.1002/cyto.990140407. [DOI] [PubMed] [Google Scholar]
  12. Katona I. M., Urban J. F., Jr, Finkelman F. D. B cells that simultaneously express surface IgM and IgE in Nippostrongylus brasiliensis-infected SJA/9 mice do not provide evidence for isotype switching without gene deletion. Proc Natl Acad Sci U S A. 1985 Jan;82(2):511–515. doi: 10.1073/pnas.82.2.511. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kinashi T., Godal T., Noma Y., Ling N. R., Yaoita Y., Honjo T. Human neoplastic B cells express more than two isotypes of immunoglobulins without deletion of heavy-chain constant-region genes. Genes Dev. 1987 Jul;1(5):465–470. doi: 10.1101/gad.1.5.465. [DOI] [PubMed] [Google Scholar]
  14. Kunimoto D. Y., Sneller M. C., Claflin L., Mushinski J. F., Strober W. Molecular analysis of double isotype expression in IgA switching. J Immunol. 1993 Feb 15;150(4):1338–1347. [PubMed] [Google Scholar]
  15. Kuritani T., Cooper M. D. Human b-cell differentiation. I. Analysis of immunoglobulin heavy chain switching using monoclonal anti-immunoglobulin M, G, and A antibodies and pokeweed mitogen-induced plasma cell differentiation. J Exp Med. 1982 Mar 1;155(3):839–851. doi: 10.1084/jem.155.3.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Linsley P. S., Bech-Hansen N. T., Siminovitch L., Cox D. W. Analysis of a break in chromosome 14 mapping to the region of the immunoglobulin heavy chain locus. Proc Natl Acad Sci U S A. 1983 Apr;80(7):1997–2001. doi: 10.1073/pnas.80.7.1997. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. MacKenzie T., Dosch H. M. Clonal and molecular characteristics of the human IgE-committed B cell subset. J Exp Med. 1989 Feb 1;169(2):407–430. doi: 10.1084/jem.169.2.407. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Manser T. The intraclonal diversity and control of antibody isotype switch recombination. Int Immunol. 1990;2(9):893–902. doi: 10.1093/intimm/2.9.893. [DOI] [PubMed] [Google Scholar]
  19. Mills F. C., Thyphronitis G., Finkelman F. D., Max E. E. Ig mu-epsilon isotype switch in IL-4-treated human B lymphoblastoid cells. Evidence for a sequential switch. J Immunol. 1992 Aug 1;149(3):1075–1085. [PubMed] [Google Scholar]
  20. Miltenyi S., Müller W., Weichel W., Radbruch A. High gradient magnetic cell separation with MACS. Cytometry. 1990;11(2):231–238. doi: 10.1002/cyto.990110203. [DOI] [PubMed] [Google Scholar]
  21. Miyawaki T., Butler J. L., Radbruch A., Gartland G. L., Cooper M. D. Isotype commitment of human B cells that are transformed by Epstein-Barr virus. Eur J Immunol. 1991 Jan;21(1):215–220. doi: 10.1002/eji.1830210132. [DOI] [PubMed] [Google Scholar]
  22. Miyawaki T., Kubagawa H., Butler J. L., Cooper M. D. Ig isotypes produced by EBV-transformed B cells as a function of age and tissue distribution. J Immunol. 1988 Jun 1;140(11):3887–3892. [PubMed] [Google Scholar]
  23. Monteiro R. C., Kubagawa H., Cooper M. D. Cellular distribution, regulation, and biochemical nature of an Fc alpha receptor in humans. J Exp Med. 1990 Mar 1;171(3):597–613. doi: 10.1084/jem.171.3.597. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Okumura K., Julius M. H., Tsu T., Herzenberg L. A., Herzenberg L. A. Demonstration that IgG memory is carried by IgG-bearing cells. Eur J Immunol. 1976 Jul;6(7):467–472. doi: 10.1002/eji.1830060704. [DOI] [PubMed] [Google Scholar]
  25. Rabbitts T. H., Forster A., Milstein C. P. Human immunoglobulin heavy chain genes: evolutionary comparisons of C mu, C delta and C gamma genes and associated switch sequences. Nucleic Acids Res. 1981 Sep 25;9(18):4509–4524. doi: 10.1093/nar/9.18.4509. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Radbruch A., Müller W., Rajewsky K. Class switch recombination is IgG1 specific on active and inactive IgH loci of IgG1-secreting B-cell blasts. Proc Natl Acad Sci U S A. 1986 Jun;83(11):3954–3957. doi: 10.1073/pnas.83.11.3954. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Radbruch A. The molecular basis of immunoglobulin class switching: switch transcription versus switch recombination. Immunol Res. 1991;10(3-4):381–385. doi: 10.1007/BF02919725. [DOI] [PubMed] [Google Scholar]
  28. Schultz C., Petrini J., Collins J., Claflin J. L., Denis K. A., Gearhart P., Gritzmacher C., Manser T., Shulman M., Dunnick W. Patterns and extent of isotype-specificity in the murine H chain switch DNA rearrangement. J Immunol. 1990 Jan 1;144(1):363–370. [PubMed] [Google Scholar]
  29. Shapira S. K., Vercelli D., Jabara H. H., Fu S. M., Geha R. S. Molecular analysis of the induction of immunoglobulin E synthesis in human B cells by interleukin 4 and engagement of CD40 antigen. J Exp Med. 1992 Jan 1;175(1):289–292. doi: 10.1084/jem.175.1.289. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Shimizu A., Honjo T. Immunoglobulin class switching. Cell. 1984 Apr;36(4):801–803. doi: 10.1016/0092-8674(84)90029-1. [DOI] [PubMed] [Google Scholar]
  31. Shimizu A., Nussenzweig M. C., Han H., Sanchez M., Honjo T. Trans-splicing as a possible molecular mechanism for the multiple isotype expression of the immunoglobulin gene. J Exp Med. 1991 Jun 1;173(6):1385–1393. doi: 10.1084/jem.173.6.1385. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Siebenkotten G., Esser C., Wabl M., Radbruch A. The murine IgG1/IgE class switch program. Eur J Immunol. 1992 Jul;22(7):1827–1834. doi: 10.1002/eji.1830220723. [DOI] [PubMed] [Google Scholar]
  33. Tavernier J., Derynck R., Fiers W. Evidence for a unique human fibroblast interferon (IFN-beta 1) chromosomal gene, devoid of intervening sequences. Nucleic Acids Res. 1981 Feb 11;9(3):461–471. doi: 10.1093/nar/9.3.461. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Thiele D. L., Kurosaka M., Lipsky P. E. Phenotype of the accessory cell necessary for mitogen-stimulated T and B cell responses in human peripheral blood: delineation by its sensitivity to the lysosomotropic agent, L-leucine methyl ester. J Immunol. 1983 Nov;131(5):2282–2290. [PubMed] [Google Scholar]
  35. Vanhove B., Bazin H. IgG2b or IgE molecules can be co-expressed with those of the IgM and IgD isotypes of the membrane of normal rat B cells. Mol Immunol. 1992 Jan;29(1):1–8. doi: 10.1016/0161-5890(92)90150-v. [DOI] [PubMed] [Google Scholar]
  36. Webb C. F., Cooper M. D., Burrows P. D., Griffin J. A. Immunoglobulin gene rearrangements and deletions in human Epstein-Barr virus-transformed cell lines producing different IgG and IgA subclasses. Proc Natl Acad Sci U S A. 1985 Aug;82(16):5495–5499. doi: 10.1073/pnas.82.16.5495. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Weiss E. A., Tucker P. W., Yuan D. The C mu gene is transcribed in IgG bearing B lymphocytes. J Mol Cell Immunol. 1987;3(2):69–81. [PubMed] [Google Scholar]
  38. Winter E., Krawinkel U., Radbruch A. Directed Ig class switch recombination in activated murine B cells. EMBO J. 1987 Jun;6(6):1663–1671. doi: 10.1002/j.1460-2075.1987.tb02415.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Yaoita Y., Kumagai Y., Okumura K., Honjo T. Expression of lymphocyte surface IgE does not require switch recombination. Nature. 1982 Jun 24;297(5868):697–699. doi: 10.1038/297697a0. [DOI] [PubMed] [Google Scholar]

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