Skip to main content
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1986 Aug 1;164(2):562–579. doi: 10.1084/jem.164.2.562

Double isotype production by a neoplastic B cell line. II. Allelically excluded production of mu and gamma 1 heavy chains without CH gene rearrangement

PMCID: PMC2188233  PMID: 3088208

Abstract

In our accompanying paper, we described a switch variant (BCL1.2.58) that expresses membrane and secreted forms of IgM and IgG1. Both IgM and IgG1 share the same idiotype and use the same VDJ rearrangement. Here, a detailed Southern blot analysis of the entire constant region of the Ig heavy chain (Ig CH) locus of parental (BCL1.B1) and variants (BCL1.B2) DNA showed no detectable rearrangement. Similar analysis of the JH-C mu region led to the conclusion that two heavy chain alleles present in the IgM/IgG1-producing variants carried the same VDJ rearrangement but differed in their 3' flanking regions. One chromosome 12 did not carry any Ig CH genes, whereas, the other chromosome 12 carried one copy of CH genes. In BCL1.B1, however, each of the chromosome 12 alleles carried a full copy of CH genes. Karyotypic analysis confirmed the presence of two translocated t(12;16) chromosomes in both BCL1.2.58 and BCL1.B1 cells, with a break 5' to the VH locus at the distal region (12F2) of chromosome 12, and at the proximal region below the centromere (16B3) of chromosome 16. We conclude that double production of IgM and IgG1 in BCL1.B2 is accomplished by transcription of the corresponding CH genes in germline configuration using a single VDJ on the same chromosome 12.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

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

  1. Alt F. W., Rosenberg N., Casanova R. J., Thomas E., Baltimore D. Immunoglobulin heavy-chain expression and class switching in a murine leukaemia cell line. Nature. 1982 Mar 25;296(5855):325–331. doi: 10.1038/296325a0. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Brooks K., Yuan D., Uhr J. W., Krammer P. H., Vitetta E. S. Lymphokine-induced IgM secretion by clones of neoplastic B cells. Nature. 1983 Apr 28;302(5911):825–826. doi: 10.1038/302825a0. [DOI] [PubMed] [Google Scholar]
  4. Burrows P. D., Beck-Engeser G. B., Wabl M. R. Immunoglobulin heavy-chain class switching in a pre-B cell line is accompanied by DNA rearrangement. Nature. 1983 Nov 17;306(5940):243–246. doi: 10.1038/306243a0. [DOI] [PubMed] [Google Scholar]
  5. Calvert J. E., Kim M. F., Gathings W. E., Cooper M. D. Differentiation of B lineage cells from liver of neonatal mice: generation of immunoglobulin isotype diversity in vitro. J Immunol. 1983 Oct;131(4):1693–1697. [PubMed] [Google Scholar]
  6. Chiorazzi N., Fu S. M., Kunkel H. G. Stimulation of human B lymphocytes by antibodies to IgM and IgG: functional evidence for the expression of IgG on B-lymphocyte surface membranes. Clin Immunol Immunopathol. 1980 Mar;15(3):301–313. doi: 10.1016/0090-1229(80)90042-2. [DOI] [PubMed] [Google Scholar]
  7. D'Eustachio P., Bothwell A. L., Takaro T. K., Baltimore D., Ruddle F. H. Chromosomal location of structural genes encoding murine immunoglobulin lambda light chains. Genetics of murine lambda light chains. J Exp Med. 1981 Apr 1;153(4):793–800. doi: 10.1084/jem.153.4.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. D'Eustachio P., Pravtcheva D., Marcu K., Ruddle F. H. Chromosomal location of the structural gene cluster encoding murine immunoglobulin heavy chains. J Exp Med. 1980 Jun 1;151(6):1545–1550. doi: 10.1084/jem.151.6.1545. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. DePinho R., Kruger K., Andrews N., Lutzker S., Baltimore D., Alt F. W. Molecular basis of heavy-chain class switching and switch region deletion in an Abelson virus-transformed cell line. Mol Cell Biol. 1984 Dec;4(12):2905–2910. doi: 10.1128/mcb.4.12.2905. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Donelson J. E., Rice-Ficht A. C. Molecular biology of trypanosome antigenic variation. Microbiol Rev. 1985 Jun;49(2):107–125. doi: 10.1128/mr.49.2.107-125.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Erikson J., Miller D. A., Miller O. J., Abcarian P. W., Skurla R. M., Mushinski J. F., Croce C. M. The c-myc oncogene is translocated to the involved chromosome 12 in mouse plasmacytoma. Proc Natl Acad Sci U S A. 1985 Jun;82(12):4212–4216. doi: 10.1073/pnas.82.12.4212. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Gilliam A. C., Shen A., Richards J. E., Blattner F. R., Mushinski J. F., Tucker P. W. Illegitimate recombination generates a class switch from C mu to C delta in an IgD-secreting plasmacytoma. Proc Natl Acad Sci U S A. 1984 Jul;81(13):4164–4168. doi: 10.1073/pnas.81.13.4164. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gronowicz E. S., Doss C. A., Howard F. D., Morrison D. C., Strober S. An in vitro line of the B cell tumor BCL1 can be activated by LPS to secrete IgM1. J Immunol. 1980 Sep;125(3):976–980. [PubMed] [Google Scholar]
  14. Honjo T., Kataoka T. Organization of immunoglobulin heavy chain genes and allelic deletion model. Proc Natl Acad Sci U S A. 1978 May;75(5):2140–2144. doi: 10.1073/pnas.75.5.2140. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Honjo T., Obata M., Yamawaki-Katoaka Y., Kataoka T., Kawakami T., Takahashi N., Mano Y. Cloning and complete nucleotide sequence of mouse immunoglobulin gamma 1 chain gene. Cell. 1979 Oct;18(2):559–568. doi: 10.1016/0092-8674(79)90072-2. [DOI] [PubMed] [Google Scholar]
  16. Isakson P. C., Uhr J. W., Krolick K. A., Finkelman F., Vitetta E. S. Acquisition of cell surface IgD after in vitro culture of neoplastic B cells from the murine tumor BCL1. J Exp Med. 1980 Mar 1;151(3):749–754. doi: 10.1084/jem.151.3.749. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. Kearney J. F., Cooper M. D., Lawton A. R. B cell differentiation induced by lipopolysaccharide. IV. Development of immunoglobulin class restriction in precursors of IgG-synthesizing cells. J Immunol. 1976 Nov;117(5 Pt 1):1567–1572. [PubMed] [Google Scholar]
  19. Keese P., Symons R. H. Domains in viroids: evidence of intermolecular RNA rearrangements and their contribution to viroid evolution. Proc Natl Acad Sci U S A. 1985 Jul;82(14):4582–4586. doi: 10.1073/pnas.82.14.4582. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Knapp M. R., Liu C. P., Newell N., Ward R. B., Tucker P. W., Strober S., Blattner F. Simultaneous expression of immunoglobulin mu and delta heavy chains by a cloned B-cell lymphoma: a single copy of the VH gene is shared by two adjacent CH genes. Proc Natl Acad Sci U S A. 1982 May;79(9):2996–3000. doi: 10.1073/pnas.79.9.2996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Liu C. P., Tucker P. W., Mushinski J. F., Blattner F. R. Mapping of heavy chain genes for mouse immunoglobulins M and D. Science. 1980 Sep 19;209(4463):1348–1353. doi: 10.1126/science.6774414. [DOI] [PubMed] [Google Scholar]
  22. Maki R., Roeder W., Traunecker A., Sidman C., Wabl M., Raschke W., Tonegawa S. The role of DNA rearrangement and alternative RNA processing in the expression of immunoglobulin delta genes. Cell. 1981 May;24(2):353–365. doi: 10.1016/0092-8674(81)90325-1. [DOI] [PubMed] [Google Scholar]
  23. Marcu K. B., Cooper M. D. New views of the immunoglobulin heavy-chain switch. Nature. 1982 Jul 22;298(5872):327–328. doi: 10.1038/298327a0. [DOI] [PubMed] [Google Scholar]
  24. Marcu K. B. Immunoglobulin heavy-chain constant-region genes. Cell. 1982 Jul;29(3):719–721. doi: 10.1016/0092-8674(82)90431-7. [DOI] [PubMed] [Google Scholar]
  25. Marcu K. B., Schibler U., Perry R. P. Nuclear transcripts of mouse heavy chain immunoglobulin genes contain only the expressed class of C-region sequences. Science. 1979 Jun 8;204(4397):1087–1088. doi: 10.1126/science.109919. [DOI] [PubMed] [Google Scholar]
  26. Mather E. L., Nelson K. J., Haimovich J., Perry R. P. Mode of regulation of immunoglobulin mu- and delta-chain expression varies during B-lymphocyte maturation. Cell. 1984 Feb;36(2):329–338. doi: 10.1016/0092-8674(84)90226-5. [DOI] [PubMed] [Google Scholar]
  27. Maxam A. M., Gilbert W. Sequencing end-labeled DNA with base-specific chemical cleavages. Methods Enzymol. 1980;65(1):499–560. doi: 10.1016/s0076-6879(80)65059-9. [DOI] [PubMed] [Google Scholar]
  28. Moore K. W., Rogers J., Hunkapiller T., Early P., Nottenburg C., Weissman I., Bazin H., Wall R., Hood L. E. Expression of IgD may use both DNA rearrangement and RNA splicing mechanisms. Proc Natl Acad Sci U S A. 1981 Mar;78(3):1800–1804. doi: 10.1073/pnas.78.3.1800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Mushinski J. F., Blattner F. R., Owens J. D., Finkelman F. D., Kessler S. W., Fitzmaurice L., Potter M., Tucker P. W. Mouse immunoglobulin D: construction and characterization of a cloned delta chain cDNA. Proc Natl Acad Sci U S A. 1980 Dec;77(12):7405–7409. doi: 10.1073/pnas.77.12.7405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Nesbitt M. N., Francke U. A system of nomenclature for band patterns of mouse chromosomes. Chromosoma. 1973;41(2):145–158. doi: 10.1007/BF00319691. [DOI] [PubMed] [Google Scholar]
  31. Newell N., Richards J. E., Tucker P. W., Blattner F. R. J genes for heavy chain immunoglobulins of mouse. Science. 1980 Sep 5;209(4461):1128–1132. doi: 10.1126/science.6250219. [DOI] [PubMed] [Google Scholar]
  32. 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]
  33. Perlmutter A. P., Gilbert W. Antibodies of the secondary response can be expressed without switch recombination in normal mouse B cells. Proc Natl Acad Sci U S A. 1984 Nov;81(22):7189–7193. doi: 10.1073/pnas.81.22.7189. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Pernis B., Forni L., Luzzati A. L. Synthesis of multiple immunoglobulin classes by single lymphocytes. Cold Spring Harb Symp Quant Biol. 1977;41(Pt 1):175–183. doi: 10.1101/sqb.1977.041.01.023. [DOI] [PubMed] [Google Scholar]
  35. Richards J. E., Gilliam A. C., Shen A., Tucker P. W., Blattner F. R. Unusual sequences in the murine immunoglobulin mu-delta heavy-chain region. Nature. 1983 Dec 1;306(5942):483–487. doi: 10.1038/306483a0. [DOI] [PubMed] [Google Scholar]
  36. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  37. Scott M. P., Weiner A. J., Hazelrigg T. I., Polisky B. A., Pirrotta V., Scalenghe F., Kaufman T. C. The molecular organization of the Antennapedia locus of Drosophila. Cell. 1983 Dec;35(3 Pt 2):763–776. doi: 10.1016/0092-8674(83)90109-5. [DOI] [PubMed] [Google Scholar]
  38. Seabright M. A rapid banding technique for human chromosomes. Lancet. 1971 Oct 30;2(7731):971–972. doi: 10.1016/s0140-6736(71)90287-x. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Shimizu A., Takahashi N., Yaoita Y., Honjo T. Organization of the constant-region gene family of the mouse immunoglobulin heavy chain. Cell. 1982 Mar;28(3):499–506. doi: 10.1016/0092-8674(82)90204-5. [DOI] [PubMed] [Google Scholar]
  41. Stavnezer J., Abbott J., Sirlin S. Immunoglobulin heavy chain switching in cultured I.29 murine B lymphoma cells: commitment to an IgA or IgE switch. Curr Top Microbiol Immunol. 1984;113:109–116. doi: 10.1007/978-3-642-69860-6_21. [DOI] [PubMed] [Google Scholar]
  42. Stavnezer J., Sirlin S., Abbott J. Induction of immunoglobulin isotype switching in cultured I.29 B lymphoma cells. Characterization of the accompanying rearrangements of heavy chain genes. J Exp Med. 1985 Mar 1;161(3):577–601. doi: 10.1084/jem.161.3.577. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Tucker P. W., Liu C. P., Mushinski J. F., Blattner F. R. Mouse immunoglobulin D: messenger RNA and genomic DNA sequences. Science. 1980 Sep 19;209(4463):1353–1360. doi: 10.1126/science.6968091. [DOI] [PubMed] [Google Scholar]
  44. Tucker P. W., Marcu K. B., Slightom J. L., Blattner F. R. Structure of the constant and 3' untranslated regions of the murine gamma 2b heavy chain messenger RNA. Science. 1979 Dec 14;206(4424):1299–1303. doi: 10.1126/science.117548. [DOI] [PubMed] [Google Scholar]
  45. 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]
  46. Vitetta E., Puré E., Isakson P., Buck L., Uhr J. The activation of murine B cells: the role of surface immunoglobulins. Immunol Rev. 1980;52:211–231. doi: 10.1111/j.1600-065x.1980.tb00336.x. [DOI] [PubMed] [Google Scholar]
  47. Voss R., Maftzir G., Slavin S. Cytogenetic analysis of the spontaneous murine B-cell leukemia (BCL1). Leuk Res. 1980;4(3):325–332. doi: 10.1016/0145-2126(80)90040-5. [DOI] [PubMed] [Google Scholar]
  48. Wels J. A., Word C. J., Rimm D., Der-Balan G. P., Martinez H. M., Tucker P. W., Blattner F. R. Structural analysis of the murine IgG3 constant region gene. EMBO J. 1984 Sep;3(9):2041–2046. doi: 10.1002/j.1460-2075.1984.tb02089.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. 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]
  50. Yuan D., Gilliam A. C., Tucker P. W. Regulation of expression of immunoglobulins M and D in murine B cells. Fed Proc. 1985 Jul;44(10):2652–2659. [PubMed] [Google Scholar]
  51. Yuan D., Tucker P. W. Transcriptional regulation of the mu-delta heavy chain locus in normal murine B lymphocytes. J Exp Med. 1984 Aug 1;160(2):564–583. doi: 10.1084/jem.160.2.564. [DOI] [PMC free article] [PubMed] [Google Scholar]
  52. Zan-Bar I., Vitetta E. S., Assisi F., Strober S. The relationship between surface immunoglobulin isotype and immune function of murine B lymphocytes. III. Expression of a single predominant isotype on primed and unprimed B cells. J Exp Med. 1978 May 1;147(5):1374–1394. doi: 10.1084/jem.147.5.1374. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES