Skip to main content
NCBI home page
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
. 1992 Aug 15;89(16):7615–7619. doi: 10.1073/pnas.89.16.7615

Stochastic rearrangement of immunoglobulin variable-region genes in chicken B-cell development.

T Benatar 1, L Tkalec 1, M J Ratcliffe 1
PMCID: PMC49761  PMID: 1502173

Abstract

The molecular mechanism by which immunoglobulin (Ig) gene rearrangement occurs is highly conserved between mammalian and avian species. However, in avian species, an equivalent to the mammalian pre-B cell, which has undergone Ig heavy-chain gene rearrangement and expresses mu heavy chains in the absence of Ig light-chain rearrangement, has not been convincingly demonstrated. It is consequently unclear whether an ordered progression of gene rearrangement events leading to functional Ig expression occurs in avian species. To examine the sequence of Ig gene rearrangement events in chicken B-cell development, we transformed day 12 embryo bursal cells with the REV-T(CSV) retrovirus. More than 100 clones were analyzed by Southern blotting and polymerase chain reaction for the presence of Ig gene rearrangements. The majority of these clones contained only germline Ig sequences. Several clones contained complete heavy- and light-chain rearrangements and 13 clones contained only heavy-chain rearrangements analogous to stages of mammalian B-cell development. However, 5 clones contained rearrangements of light-chain genes in the absence of complete heavy-chain rearrangement. Consequently, we conclude that rearrangement of chicken Ig light-chain genes does not require heavy-chain variable-region rearrangement. This observation suggests that chicken Ig gene rearrangement events required for Ig expression occur stochastically rather than sequentially.

Full text

PDF
7615

Images in this article

7616Image
on p.7616
7617Image
on p.7617

Selected References

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

  1. Barth C. F., Humphries E. H. A nonimmunosuppressive helper virus allows high efficiency induction of B cell lymphomas by reticuloendotheliosis virus strain T. J Exp Med. 1988 Jan 1;167(1):89–108. doi: 10.1084/jem.167.1.89. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Barth C. F., Humphries E. H. Expression of v-rel induces mature B-cell lines that reflect the diversity of avian immunoglobulin heavy- and light-chain rearrangements. Mol Cell Biol. 1988 Dec;8(12):5358–5368. doi: 10.1128/mcb.8.12.5358. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Benatar T., Iacampo S., Tkalec L., Ratcliffe M. J. Expression of immunoglobulin genes in the avian embryo bone marrow revealed by retroviral transformation. Eur J Immunol. 1991 Oct;21(10):2529–2536. doi: 10.1002/eji.1830211033. [DOI] [PubMed] [Google Scholar]
  4. Bucchini D., Reynaud C. A., Ripoche M. A., Grimal H., Jami J., Weill J. C. Rearrangement of a chicken immunoglobulin gene occurs in the lymphoid lineage of transgenic mice. 1987 Mar 26-Apr 1Nature. 326(6111):409–411. doi: 10.1038/326409a0. [DOI] [PubMed] [Google Scholar]
  5. Chen L., Lim M. Y., Bose H., Jr, Bishop J. M. Rearrangements of chicken immunoglobulin genes in lymphoid cells transformed by the avian retroviral oncogene v-rel. Proc Natl Acad Sci U S A. 1988 Jan;85(2):549–553. doi: 10.1073/pnas.85.2.549. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper M. D., Cain W. A., Van Alten P. J., Good R. A. Development and function of the immunoglobulin producing system. I. Effect of bursectomy at different stages of development on germinal centers, plasma cells, immunoglobulins and antibody production. Int Arch Allergy Appl Immunol. 1969;35(3):242–252. [PubMed] [Google Scholar]
  7. Grossi C. E., Lydyard P. M., Cooper M. D. Ontogeny of B cells in the chicken. II. Changing patterns of cytoplasmic IgM expression and of modulation requirements for surface IgM by anti-mu antibodies. J Immunol. 1977 Aug;119(2):749–756. [PubMed] [Google Scholar]
  8. Houssaint E., Belo M., Le Douarin N. M. Investigations on cell lineage and tissue interactions in the developing bursa of Fabricius through interspecific chimeras. Dev Biol. 1976 Oct 15;53(2):250–264. doi: 10.1016/0012-1606(76)90227-x. [DOI] [PubMed] [Google Scholar]
  9. Hozumi N., Tonegawa S. Evidence for somatic rearrangement of immunoglobulin genes coding for variable and constant regions. Proc Natl Acad Sci U S A. 1976 Oct;73(10):3628–3632. doi: 10.1073/pnas.73.10.3628. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Iglesias A., Kopf M., Williams G. S., Bühler B., Köhler G. Molecular requirements for the mu-induced light chain gene rearrangement in pre-B cells. EMBO J. 1991 Aug;10(8):2147–2155. doi: 10.1002/j.1460-2075.1991.tb07749.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Jalkanen S., Granfors K., Jalkanen M., Toivanen P. Immune capacity of the chicken bursectomized at 60 hours of incubation: failure to produce immune, natural, and autoantibodies in spite of immunoglobulin production. Cell Immunol. 1983 Sep;80(2):363–373. doi: 10.1016/0008-8749(83)90124-7. [DOI] [PubMed] [Google Scholar]
  12. Kubagawa H., Cooper M. D., Carroll A. J., Burrows P. D. Light-chain gene expression before heavy-chain gene rearrangement in pre-B cells transformed by Epstein-Barr virus. Proc Natl Acad Sci U S A. 1989 Apr;86(7):2356–2360. doi: 10.1073/pnas.86.7.2356. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Kurosawa Y., von Boehmer H., Haas W., Sakano H., Trauneker A., Tonegawa S. Identification of D segments of immunoglobulin heavy-chain genes and their rearrangement in T lymphocytes. Nature. 1981 Apr 16;290(5807):565–570. doi: 10.1038/290565a0. [DOI] [PubMed] [Google Scholar]
  14. Mansikka A., Jalkanen S., Sandberg M., Granfors K., Lassila O., Toivanen P. Bursectomy of chicken embryos at 60 hours of incubation leads to an oligoclonal B cell compartment and restricted Ig diversity. J Immunol. 1990 Dec 1;145(11):3601–3609. [PubMed] [Google Scholar]
  15. McCormack W. T., Tjoelker L. W., Barth C. F., Carlson L. M., Petryniak B., Humphries E. H., Thompson C. B. Selection for B cells with productive IgL gene rearrangements occurs in the bursa of Fabricius during chicken embryonic development. Genes Dev. 1989 Jun;3(6):838–847. doi: 10.1101/gad.3.6.838. [DOI] [PubMed] [Google Scholar]
  16. McCormack W. T., Tjoelker L. W., Carlson L. M., Petryniak B., Barth C. F., Humphries E. H., Thompson C. B. Chicken IgL gene rearrangement involves deletion of a circular episome and addition of single nonrandom nucleotides to both coding segments. Cell. 1989 Mar 10;56(5):785–791. doi: 10.1016/0092-8674(89)90683-1. [DOI] [PubMed] [Google Scholar]
  17. Moore M. A., Owen J. J. Chromosome marker studies on the development of the haemopoietic system in the chick embryo. Nature. 1965 Dec 4;208(5014):956–passim. doi: 10.1038/208956a0. [DOI] [PubMed] [Google Scholar]
  18. Mussman H. C., Twiehaus M. J. Pathogenesis of reticuloendothelial virus disease in chicks--an acute runting syndrome. Avian Dis. 1971 Jul-Sep;15(3):483–502. [PubMed] [Google Scholar]
  19. Osmond D. G. Population dynamics of bone marrow B lymphocytes. Immunol Rev. 1986 Oct;93:103–124. doi: 10.1111/j.1600-065x.1986.tb01504.x. [DOI] [PubMed] [Google Scholar]
  20. Pink J. R. A comment on allelic exclusion, 'D-disaster' and germ-line gene specificities. Immunol Rev. 1990 Jun;115:239–242. doi: 10.1111/j.1600-065x.1990.tb00794.x. [DOI] [PubMed] [Google Scholar]
  21. Ramsden D. A., Wu G. E. Mouse kappa light-chain recombination signal sequences mediate recombination more frequently than do those of lambda light chain. Proc Natl Acad Sci U S A. 1991 Dec 1;88(23):10721–10725. doi: 10.1073/pnas.88.23.10721. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Ratcliffe M. J., Lassila O., Pink J. R., Vainio O. Avian B cell precursors: surface immunoglobulin expression is an early, possibly bursa-independent event. Eur J Immunol. 1986 Feb;16(2):129–133. doi: 10.1002/eji.1830160204. [DOI] [PubMed] [Google Scholar]
  23. Ratcliffe M. J., Paramithiotis E. The end can justify the means. Semin Immunol. 1990 May;2(3):217–226. [PubMed] [Google Scholar]
  24. Reth M. G., Ammirati P., Jackson S., Alt F. W. Regulated progression of a cultured pre-B-cell line to the B-cell stage. 1985 Sep 26-Oct 2Nature. 317(6035):353–355. doi: 10.1038/317353a0. [DOI] [PubMed] [Google Scholar]
  25. Reth M., Petrac E., Wiese P., Lobel L., Alt F. W. Activation of V kappa gene rearrangement in pre-B cells follows the expression of membrane-bound immunoglobulin heavy chains. EMBO J. 1987 Nov;6(11):3299–3305. doi: 10.1002/j.1460-2075.1987.tb02649.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Reynaud C. A., Anquez V., Dahan A., Weill J. C. A single rearrangement event generates most of the chicken immunoglobulin light chain diversity. Cell. 1985 Feb;40(2):283–291. doi: 10.1016/0092-8674(85)90142-4. [DOI] [PubMed] [Google Scholar]
  27. Reynaud C. A., Anquez V., Grimal H., Weill J. C. A hyperconversion mechanism generates the chicken light chain preimmune repertoire. Cell. 1987 Feb 13;48(3):379–388. doi: 10.1016/0092-8674(87)90189-9. [DOI] [PubMed] [Google Scholar]
  28. Reynaud C. A., Anquez V., Weill J. C. The chicken D locus and its contribution to the immunoglobulin heavy chain repertoire. Eur J Immunol. 1991 Nov;21(11):2661–2670. doi: 10.1002/eji.1830211104. [DOI] [PubMed] [Google Scholar]
  29. Reynaud C. A., Dahan A., Anquez V., Weill J. C. Somatic hyperconversion diversifies the single Vh gene of the chicken with a high incidence in the D region. Cell. 1989 Oct 6;59(1):171–183. doi: 10.1016/0092-8674(89)90879-9. [DOI] [PubMed] [Google Scholar]
  30. Thompson C. B., Neiman P. E. Somatic diversification of the chicken immunoglobulin light chain gene is limited to the rearranged variable gene segment. Cell. 1987 Feb 13;48(3):369–378. doi: 10.1016/0092-8674(87)90188-7. [DOI] [PubMed] [Google Scholar]
  31. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  32. Weber W. T., Foglia L. M. Evidence for the presence of precursor B cells in normal and in hormonally bursectomized chick embryos. Cell Immunol. 1980 Jun;52(1):84–94. doi: 10.1016/0008-8749(80)90402-5. [DOI] [PubMed] [Google Scholar]
  33. Weill J. C., Reynaud C. A., Lassila O., Pink J. R. Rearrangement of chicken immunoglobulin genes is not an ongoing process in the embryonic bursa of Fabricius. Proc Natl Acad Sci U S A. 1986 May;83(10):3336–3340. doi: 10.1073/pnas.83.10.3336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Zhang J. Y., Bargmann W., Bose H. R., Jr Rearrangement and diversification of immunoglobulin light-chain genes in lymphoid cells transformed by reticuloendotheliosis virus. Mol Cell Biol. 1989 Nov;9(11):4970–4976. doi: 10.1128/mcb.9.11.4970. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Zhang J. Y., Olson W., Ewert D., Bargmann W., Bose H. R., Jr The v-rel oncogene of avian reticuloendotheliosis virus transforms immature and mature lymphoid cells of the B cell lineage in vitro. Virology. 1991 Aug;183(2):457–466. doi: 10.1016/0042-6822(91)90975-h. [DOI] [PubMed] [Google Scholar]

Articles from Proceedings of the National Academy of Sciences of the United States of America are provided here courtesy of National Academy of Sciences

RESOURCES