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The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Jan 15;97(2):477–485. doi: 10.1172/JCI118438

IgA class switch in I alpha exon-deficient mice. Role of germline transcription in class switch recombination.

G R Harriman 1, A Bradley 1, S Das 1, P Rogers-Fani 1, A C Davis 1
PMCID: PMC507040  PMID: 8567970

Abstract

Studies have implicated defective Ig class switch in the pathogenesis of IgA deficiency. To understand better the molecular events that regulate IgA class switch, a 1.4-kb region of the IgA locus containing the I alpha exon was replaced with a human hypoxanthine phosphoribosyltransferase minigene by gene targeting in murine embryonic stem cells. The I alpha exon-deficient mice derived from these embryonic stem cells had normal IgA levels in serum and secretions and normal numbers of IgA B cells in Peyer's patches and spleen. Further, I alpha exon-deficient B cells efficiently underwent IgA class switch in vitro, despite the absence of I alpha exon-containing germline transcripts. Notably, I alpha exon-deficient B cells did not require TGF-beta for IgA class switch since stimulation with LPS alone led to IgA expression. Nonetheless, whereas I alpha exon-deficient B cells constitutively expressed human hypoxanthine phosphoribosyltransferase transcripts, they did not produce IgA in the absence of LPS stimulation. These results demonstrate that the I alpha exon or transcripts containing the I alpha exon are not required for IgA class switch. Further, the effects of TGF-beta on I alpha locus transcription can be supplanted by expression of a heterologous minigene at that locus, but a second signal is required for the induction of IgA class switch.

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Selected References

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

  1. Allen R. C., Armitage R. J., Conley M. E., Rosenblatt H., Jenkins N. A., Copeland N. G., Bedell M. A., Edelhoff S., Disteche C. M., Simoneaux D. K. CD40 ligand gene defects responsible for X-linked hyper-IgM syndrome. Science. 1993 Feb 12;259(5097):990–993. doi: 10.1126/science.7679801. [DOI] [PubMed] [Google Scholar]
  2. Alt F. W., Blackwell T. K., DePinho R. A., Reth M. G., Yancopoulos G. D. Regulation of genome rearrangement events during lymphocyte differentiation. Immunol Rev. 1986 Feb;89:5–30. doi: 10.1111/j.1600-065x.1986.tb01470.x. [DOI] [PubMed] [Google Scholar]
  3. Aruffo A., Farrington M., Hollenbaugh D., Li X., Milatovich A., Nonoyama S., Bajorath J., Grosmaire L. S., Stenkamp R., Neubauer M. The CD40 ligand, gp39, is defective in activated T cells from patients with X-linked hyper-IgM syndrome. Cell. 1993 Jan 29;72(2):291–300. doi: 10.1016/0092-8674(93)90668-g. [DOI] [PubMed] [Google Scholar]
  4. Bottaro A., Lansford R., Xu L., Zhang J., Rothman P., Alt F. W. S region transcription per se promotes basal IgE class switch recombination but additional factors regulate the efficiency of the process. EMBO J. 1994 Feb 1;13(3):665–674. doi: 10.1002/j.1460-2075.1994.tb06305.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Butcher E. C., Rouse R. V., Coffman R. L., Nottenburg C. N., Hardy R. R., Weissman I. L. Surface phenotype of Peyer's patch germinal center cells: implications for the role of germinal centers in B cell differentiation. J Immunol. 1982 Dec;129(6):2698–2707. [PubMed] [Google Scholar]
  6. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  7. Coffman R. L., Lebman D. A., Shrader B. Transforming growth factor beta specifically enhances IgA production by lipopolysaccharide-stimulated murine B lymphocytes. J Exp Med. 1989 Sep 1;170(3):1039–1044. doi: 10.1084/jem.170.3.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Defrance T., Vanbervliet B., Brière F., Durand I., Rousset F., Banchereau J. Interleukin 10 and transforming growth factor beta cooperate to induce anti-CD40-activated naive human B cells to secrete immunoglobulin A. J Exp Med. 1992 Mar 1;175(3):671–682. doi: 10.1084/jem.175.3.671. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DiSanto J. P., Bonnefoy J. Y., Gauchat J. F., Fischer A., de Saint Basile G. CD40 ligand mutations in x-linked immunodeficiency with hyper-IgM. Nature. 1993 Feb 11;361(6412):541–543. doi: 10.1038/361541a0. [DOI] [PubMed] [Google Scholar]
  10. Ehrhardt R. O., Strober W., Harriman G. R. Effect of transforming growth factor (TGF)-beta 1 on IgA isotype expression. TGF-beta 1 induces a small increase in sIgA+ B cells regardless of the method of B cell activation. J Immunol. 1992 Jun 15;148(12):3830–3836. [PubMed] [Google Scholar]
  11. Elson C. O., Ealding W., Lefkowitz J. A lavage technique allowing repeated measurement of IgA antibody in mouse intestinal secretions. J Immunol Methods. 1984 Feb 24;67(1):101–108. doi: 10.1016/0022-1759(84)90089-9. [DOI] [PubMed] [Google Scholar]
  12. 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]
  13. Fuleihan R., Ramesh N., Loh R., Jabara H., Rosen R. S., Chatila T., Fu S. M., Stamenkovic I., Geha R. S. Defective expression of the CD40 ligand in X chromosome-linked immunoglobulin deficiency with normal or elevated IgM. Proc Natl Acad Sci U S A. 1993 Mar 15;90(6):2170–2173. doi: 10.1073/pnas.90.6.2170. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gaff C., Gerondakis S. RNA splicing generates alternate forms of germline immunoglobulin alpha heavy chain transcripts. Int Immunol. 1990;2(12):1143–1148. doi: 10.1093/intimm/2.12.1143. [DOI] [PubMed] [Google Scholar]
  15. Han H., Okamoto M., Honjo T., Shimizu A. Regulated expression of immunoglobulin trans-mRNA consisting of the variable region of a transgenic mu chain and constant regions of endogenous isotypes. Int Immunol. 1991 Dec;3(12):1197–1206. doi: 10.1093/intimm/3.12.1197. [DOI] [PubMed] [Google Scholar]
  16. Harriman G. R., Kunimoto D. Y., Elliott J. F., Paetkau V., Strober W. The role of IL-5 in IgA B cell differentiation. J Immunol. 1988 May 1;140(9):3033–3039. [PubMed] [Google Scholar]
  17. Harriman W., Völk H., Defranoux N., Wabl M. Immunoglobulin class switch recombination. Annu Rev Immunol. 1993;11:361–384. doi: 10.1146/annurev.iy.11.040193.002045. [DOI] [PubMed] [Google Scholar]
  18. Islam K. B., Baskin B., Nilsson L., Hammarström L., Sideras P., Smith C. I. Molecular analysis of IgA deficiency. Evidence for impaired switching to IgA. J Immunol. 1994 Feb 1;152(3):1442–1452. [PubMed] [Google Scholar]
  19. Jung S., Rajewsky K., Radbruch A. Shutdown of class switch recombination by deletion of a switch region control element. Science. 1993 Feb 12;259(5097):984–987. doi: 10.1126/science.8438159. [DOI] [PubMed] [Google Scholar]
  20. Kim P. H., Kagnoff M. F. Transforming growth factor-beta 1 is a costimulator for IgA production. J Immunol. 1990 May 1;144(9):3411–3416. [PubMed] [Google Scholar]
  21. Korthäuer U., Graf D., Mages H. W., Brière F., Padayachee M., Malcolm S., Ugazio A. G., Notarangelo L. D., Levinsky R. J., Kroczek R. A. Defective expression of T-cell CD40 ligand causes X-linked immunodeficiency with hyper-IgM. Nature. 1993 Feb 11;361(6412):539–541. doi: 10.1038/361539a0. [DOI] [PubMed] [Google Scholar]
  22. Kunimoto D. Y., Allison K. C., Watson C., Fuerst T., Armstrong G. D., Paul W., Strober W. High-level production of murine interleukin-5 (IL-5) utilizing recombinant baculovirus expression. Purification of the rIL-5 and its use in assessing the biologic role of IL-5 glycosylation. Cytokine. 1991 May;3(3):224–230. doi: 10.1016/1043-4666(91)90020-e. [DOI] [PubMed] [Google Scholar]
  23. Lebman D. A., Lee F. D., Coffman R. L. Mechanism for transforming growth factor beta and IL-2 enhancement of IgA expression in lipopolysaccharide-stimulated B cell cultures. J Immunol. 1990 Feb 1;144(3):952–959. [PubMed] [Google Scholar]
  24. Lebman D. A., Nomura D. Y., Coffman R. L., Lee F. D. Molecular characterization of germ-line immunoglobulin A transcripts produced during transforming growth factor type beta-induced isotype switching. Proc Natl Acad Sci U S A. 1990 May;87(10):3962–3966. doi: 10.1073/pnas.87.10.3962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Leung H., Maizels N. Transcriptional regulatory elements stimulate recombination in extrachromosomal substrates carrying immunoglobulin switch-region sequences. Proc Natl Acad Sci U S A. 1992 May 1;89(9):4154–4158. doi: 10.1073/pnas.89.9.4154. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lin Y. C., Stavnezer J. Regulation of transcription of the germ-line Ig alpha constant region gene by an ATF element and by novel transforming growth factor-beta 1-responsive elements. J Immunol. 1992 Nov 1;149(9):2914–2925. [PubMed] [Google Scholar]
  27. Lorenz M., Jung S., Radbruch A. Switch transcripts in immunoglobulin class switching. Science. 1995 Mar 24;267(5205):1825–1828. doi: 10.1126/science.7892607. [DOI] [PubMed] [Google Scholar]
  28. Lutzker S., Rothman P., Pollock R., Coffman R., Alt F. W. Mitogen- and IL-4-regulated expression of germ-line Ig gamma 2b transcripts: evidence for directed heavy chain class switching. Cell. 1988 Apr 22;53(2):177–184. doi: 10.1016/0092-8674(88)90379-0. [DOI] [PubMed] [Google Scholar]
  29. Marcu K. B., Banerji J., Penncavage N. A., Lang R., Arnheim N. 5' flanking region of immunoglobulin heavy chain constant region genes displays length heterogeneity in germlines of inbred mouse strains. Cell. 1980 Nov;22(1 Pt 1):187–196. doi: 10.1016/0092-8674(80)90167-1. [DOI] [PubMed] [Google Scholar]
  30. Matzuk M. M., Finegold M. J., Su J. G., Hsueh A. J., Bradley A. Alpha-inhibin is a tumour-suppressor gene with gonadal specificity in mice. Nature. 1992 Nov 26;360(6402):313–319. doi: 10.1038/360313a0. [DOI] [PubMed] [Google Scholar]
  31. McBurney M. W., Sutherland L. C., Adra C. N., Leclair B., Rudnicki M. A., Jardine K. The mouse Pgk-1 gene promoter contains an upstream activator sequence. Nucleic Acids Res. 1991 Oct 25;19(20):5755–5761. doi: 10.1093/nar/19.20.5755. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. McMahon A. P., Bradley A. The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell. 1990 Sep 21;62(6):1073–1085. doi: 10.1016/0092-8674(90)90385-r. [DOI] [PubMed] [Google Scholar]
  33. Nishida Y., Kataoka T., Ishida N., Nakai S., Kishimoto T., Böttcher I., Honjo T. Cloning of mouse immunoglobulin epsilon gene and its location within the heavy chain gene cluster. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1581–1585. doi: 10.1073/pnas.78.3.1581. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Radcliffe G., Lin Y. C., Julius M., Marcu K. B., Stavnezer J. Structure of germ line immunoglobulin alpha heavy-chain RNA and its location on polysomes. Mol Cell Biol. 1990 Jan;10(1):382–386. doi: 10.1128/mcb.10.1.382. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Ramírez-Solis R., Rivera-Pérez J., Wallace J. D., Wims M., Zheng H., Bradley A. Genomic DNA microextraction: a method to screen numerous samples. Anal Biochem. 1992 Mar;201(2):331–335. doi: 10.1016/0003-2697(92)90347-a. [DOI] [PubMed] [Google Scholar]
  36. Reaban M. E., Griffin J. A. Induction of RNA-stabilized DNA conformers by transcription of an immunoglobulin switch region. Nature. 1990 Nov 22;348(6299):342–344. doi: 10.1038/348342a0. [DOI] [PubMed] [Google Scholar]
  37. Severinson E., Fernandez C., Stavnezer J. Induction of germ-line immunoglobulin heavy chain transcripts by mitogens and interleukins prior to switch recombination. Eur J Immunol. 1990 May;20(5):1079–1084. doi: 10.1002/eji.1830200520. [DOI] [PubMed] [Google Scholar]
  38. Shimizu A., Honjo T. Synthesis and regulation of trans-mRNA encoding the immunoglobulin epsilon heavy chain. FASEB J. 1993 Jan;7(1):149–154. doi: 10.1096/fasebj.7.1.7916698. [DOI] [PubMed] [Google Scholar]
  39. Shimizu A., Kinashi T., Nussenzweig M. C., Mizuta T. R., Leder P., Honjo T. Molecular mechanism for immunoglobulin double-isotype expression. Cold Spring Harb Symp Quant Biol. 1989;54(Pt 1):175–181. doi: 10.1101/sqb.1989.054.01.022. [DOI] [PubMed] [Google Scholar]
  40. 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]
  41. Shimizu A., Nussenzweig M. C., Mizuta T. R., Leder P., Honjo T. Immunoglobulin double-isotype expression by trans-mRNA in a human immunoglobulin transgenic mouse. Proc Natl Acad Sci U S A. 1989 Oct;86(20):8020–8023. doi: 10.1073/pnas.86.20.8020. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Sonoda E., Matsumoto R., Hitoshi Y., Ishii T., Sugimoto M., Araki S., Tominaga A., Yamaguchi N., Takatsu K. Transforming growth factor beta induces IgA production and acts additively with interleukin 5 for IgA production. J Exp Med. 1989 Oct 1;170(4):1415–1420. doi: 10.1084/jem.170.4.1415. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Soriano P., Montgomery C., Geske R., Bradley A. Targeted disruption of the c-src proto-oncogene leads to osteopetrosis in mice. Cell. 1991 Feb 22;64(4):693–702. doi: 10.1016/0092-8674(91)90499-o. [DOI] [PubMed] [Google Scholar]
  44. Southern E. M. Detection of specific sequences among DNA fragments separated by gel electrophoresis. J Mol Biol. 1975 Nov 5;98(3):503–517. doi: 10.1016/s0022-2836(75)80083-0. [DOI] [PubMed] [Google Scholar]
  45. Stavnezer J., Radcliffe G., Lin Y. C., Nietupski J., Berggren L., Sitia R., Severinson E. Immunoglobulin heavy-chain switching may be directed by prior induction of transcripts from constant-region genes. Proc Natl Acad Sci U S A. 1988 Oct;85(20):7704–7708. doi: 10.1073/pnas.85.20.7704. [DOI] [PMC free article] [PubMed] [Google Scholar]
  46. Strober W., Sneller M. C. IgA deficiency. Ann Allergy. 1991 May;66(5):363–375. [PubMed] [Google Scholar]
  47. Tucker P. W., Slightom J. L., Blattner F. R. Mouse IgA heavy chain gene sequence: implications for evolution of immunoglobulin hinge axons. Proc Natl Acad Sci U S A. 1981 Dec;78(12):7684–7688. doi: 10.1073/pnas.78.12.7684. [DOI] [PMC free article] [PubMed] [Google Scholar]
  48. Yang T. P., Singer-Sam J., Flores J. C., Riggs A. D. DNA binding factors for the CpG-rich island containing the promoter of the human X-linked PGK gene. Somat Cell Mol Genet. 1988 Sep;14(5):461–472. doi: 10.1007/BF01534712. [DOI] [PubMed] [Google Scholar]
  49. Zhang J., Bottaro A., Li S., Stewart V., Alt F. W. A selective defect in IgG2b switching as a result of targeted mutation of the I gamma 2b promoter and exon. EMBO J. 1993 Sep;12(9):3529–3537. doi: 10.1002/j.1460-2075.1993.tb06027.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. van Vlasselaer P., Punnonen J., de Vries J. E. Transforming growth factor-beta directs IgA switching in human B cells. J Immunol. 1992 Apr 1;148(7):2062–2067. [PubMed] [Google Scholar]

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