Skip to main content
NCBI home page
Nucleic Acids Research logoLink to Nucleic Acids Research
. 1983 Jun 25;11(12):4007–4017. doi: 10.1093/nar/11.12.4007

Immunoglobulin diversity: analysis of the germ-line VH gene repertoire of the murine anti-GAT response.

C Schiff, M Milili, M Fougereau
PMCID: PMC326021  PMID: 6306571

Abstract

A cDNA clone was constructed from a mRNA encoding an anti-GAT (Glu60 Ala30 Tyr10) BALB/c monoclonal antibody heavy chain. Its sequence, covering codons -5 to 162 and therefore encompassing the complete V-D-J region, was determined. Surprisingly, the sequence of the VH gene-encoded region was almost identical with that of the BALB/c VH anti-HNP (4-hydroxy-3-nitrophenyl) acetyl VH region, suggesting that the same VH germ-line might be used to encode two heavy chains contributing to antibodies of discrete specificities. A specific VH probe was derived and annealed to Eco RI and Bg1 II restriction fragments of liver (unrearranged) DNA extracted from the BALB/c, DBA/2 and C57BL/6 mouse strains that differ in their H chain allotypes. Under stringent conditions, only a few bands were identified by Southern blotting. The different patterns observed suggest that the VH anti-GAT repertoire differs between these strains even though their various anti-GAT antibodies express the same public idiotypic specificities.

Full text

PDF
4007

Images in this article

4012Image
on p.4012

Selected References

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

  1. Adams J. M., Kemp D. J., Bernard O., Gough N., Webb E., Tyler B., Gerondakis S., Cory S. Organization and expression of murine immunoglobulin genes. Immunol Rev. 1981;59:5–32. doi: 10.1111/j.1600-065x.1981.tb00454.x. [DOI] [PubMed] [Google Scholar]
  2. Amzel L. M., Poljak R. J., Saul F., Varga J. M., Richards F. F. The three dimensional structure of a combining region-ligand complex of immunoglobulin NEW at 3.5-A resolution. Proc Natl Acad Sci U S A. 1974 Apr;71(4):1427–1430. doi: 10.1073/pnas.71.4.1427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Auffray C., Rougeon F. Purification of mouse immunoglobulin heavy-chain messenger RNAs from total myeloma tumor RNA. Eur J Biochem. 1980 Jun;107(2):303–314. doi: 10.1111/j.1432-1033.1980.tb06030.x. [DOI] [PubMed] [Google Scholar]
  4. Ben-Neriah Y., Cohen J. B., Rechavi G., Zakut R., Givol D. Polymorphism of germ-line immunoglobulin VH genes correlates with allotype and idiotype markers. Eur J Immunol. 1981 Dec;11(12):1017–1020. doi: 10.1002/eji.1830111213. [DOI] [PubMed] [Google Scholar]
  5. Bentley D. L., Rabbitts T. H. Human V kappa immunoglobulin gene number: implications for the origin of antibody diversity. Cell. 1981 Jun;24(3):613–623. doi: 10.1016/0092-8674(81)90088-x. [DOI] [PubMed] [Google Scholar]
  6. Birnboim H. C., Doly J. A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucleic Acids Res. 1979 Nov 24;7(6):1513–1523. doi: 10.1093/nar/7.6.1513. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a gamma 2a variable region. Cell. 1981 Jun;24(3):625–637. doi: 10.1016/0092-8674(81)90089-1. [DOI] [PubMed] [Google Scholar]
  8. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Heavy chain variable region contribution to the NPb family of antibodies: somatic mutation evident in a gamma 2a variable region. Cell. 1981 Jun;24(3):625–637. doi: 10.1016/0092-8674(81)90089-1. [DOI] [PubMed] [Google Scholar]
  9. Brack C., Hirama M., Lenhard-Schuller R., Tonegawa S. A complete immunoglobulin gene is created by somatic recombination. Cell. 1978 Sep;15(1):1–14. doi: 10.1016/0092-8674(78)90078-8. [DOI] [PubMed] [Google Scholar]
  10. Buell G. N., Wickens M. P., Payvar F., Schimke R. T. Synthesis of full length cDNAs from four partially purified oviduct mRNAs. J Biol Chem. 1978 Apr 10;253(7):2471–2482. [PubMed] [Google Scholar]
  11. Crews S., Griffin J., Huang H., Calame K., Hood L. A single VH gene segment encodes the immune response to phosphorylcholine: somatic mutation is correlated with the class of the antibody. Cell. 1981 Jul;25(1):59–66. doi: 10.1016/0092-8674(81)90231-2. [DOI] [PubMed] [Google Scholar]
  12. Drouin J. Cloning of human mitochondrial DNA in Escherichia coli. J Mol Biol. 1980 Jun 15;140(1):15–34. doi: 10.1016/0022-2836(80)90354-x. [DOI] [PubMed] [Google Scholar]
  13. Early P., Huang H., Davis M., Calame K., Hood L. An immunoglobulin heavy chain variable region gene is generated from three segments of DNA: VH, D and JH. Cell. 1980 Apr;19(4):981–992. doi: 10.1016/0092-8674(80)90089-6. [DOI] [PubMed] [Google Scholar]
  14. Efstratiadis A., Kafatos F. C., Maxam A. M., Maniatis T. Enzymatic in vitro synthesis of globin genes. Cell. 1976 Feb;7(2):279–288. doi: 10.1016/0092-8674(76)90027-1. [DOI] [PubMed] [Google Scholar]
  15. Gearhart P. J., Johnson N. D., Douglas R., Hood L. IgG antibodies to phosphorylcholine exhibit more diversity than their IgM counterparts. Nature. 1981 May 7;291(5810):29–34. doi: 10.1038/291029a0. [DOI] [PubMed] [Google Scholar]
  16. Givol D., Zakut R., Effron K., Rechavi G., Ram D., Cohen J. B. Diversity of germ-line immunoglobulin VH genes. Nature. 1981 Jul 30;292(5822):426–430. doi: 10.1038/292426a0. [DOI] [PubMed] [Google Scholar]
  17. Gross-Bellard M., Oudet P., Chambon P. Isolation of high-molecular-weight DNA from mammalian cells. Eur J Biochem. 1973 Jul 2;36(1):32–38. doi: 10.1111/j.1432-1033.1973.tb02881.x. [DOI] [PubMed] [Google Scholar]
  18. Hamlyn P. H., Browniee G. G., Cheng C. C., Gait M. J., Milstein C. Complete sequence of constant and 3' noncoding regions of an immunoglobulin mRNA using the dideoxynucleotide method of RNA sequencing. Cell. 1978 Nov;15(3):1067–1075. doi: 10.1016/0092-8674(78)90290-8. [DOI] [PubMed] [Google Scholar]
  19. Heidmann O., Rougeon F. Multiple sequences related to a constant-region kappa light chain gene in the rabbit genome. Cell. 1982 Mar;28(3):507–513. doi: 10.1016/0092-8674(82)90205-7. [DOI] [PubMed] [Google Scholar]
  20. Ju S. T., Kipps T. J., Theze J., Benacerraf B., Dorf M. E. Idiotypic analysis of anti-GAT antibodies. I. Presence of common idiotypic specificities in both responder and nonresponder mice. J Immunol. 1978 Sep;121(3):1034–1039. [PubMed] [Google Scholar]
  21. Kearney J. F., Radbruch A., Liesegang B., Rajewsky K. A new mouse myeloma cell line that has lost immunoglobulin expression but permits the construction of antibody-secreting hybrid cell lines. J Immunol. 1979 Oct;123(4):1548–1550. [PubMed] [Google Scholar]
  22. Kurosawa Y., Tonegawa S. Organization, structure, and assembly of immunoglobulin heavy chain diversity DNA segments. J Exp Med. 1982 Jan 1;155(1):201–218. doi: 10.1084/jem.155.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Leclercq L., Mazié J. C., Sommé G., Thèze J. Monoclonal anti-GAT antibodies with different fine specificities express the same public idiotype. Mol Immunol. 1982 Aug;19(8):1001–1009. doi: 10.1016/0161-5890(82)90308-x. [DOI] [PubMed] [Google Scholar]
  24. Loh D. Y., Bothwell A. L., White-Scharf M. E., Imanishi-Kari T., Baltimore D. Molecular basis of a mouse strain-specific anti-hapten response. Cell. 1983 May;33(1):85–93. doi: 10.1016/0092-8674(83)90337-9. [DOI] [PubMed] [Google Scholar]
  25. 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]
  26. Obata M., Yamawaki-Kataoka Y., Takahashi N., Kataoka T., Shimizu A., Mano Y., Seidman J. G., Peterlin B. M., Leder P., Honjo T. Immunoglobulin gamma 1 heavy chain gene: structural gene sequences cloned in a bacterial plasmid. Gene. 1980 Apr;9(1-2):87–97. doi: 10.1016/0378-1119(80)90168-7. [DOI] [PubMed] [Google Scholar]
  27. Rabbitts T. H., Matthyssens G., Hamlyn P. H. Contribution of immunoglobulin heavy-chain variable-region genes to antibody diversity. Nature. 1980 Mar 20;284(5753):238–243. doi: 10.1038/284238a0. [DOI] [PubMed] [Google Scholar]
  28. Retzel E. F., Collett M. S., Faras A. J. Enzymatic synthesis of deoxyribonucleic acid by the avian retrovirus reverse transcriptase in vitro: optimum conditions required for transcription of large ribonucleic acid templates. Biochemistry. 1980 Feb 5;19(3):513–518. doi: 10.1021/bi00544a019. [DOI] [PubMed] [Google Scholar]
  29. 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]
  30. Roskam W. G., Rougeon F. Molecular cloning and nucleotide sequence of the human growth hormone structural gene. Nucleic Acids Res. 1979 Sep 25;7(2):305–320. doi: 10.1093/nar/7.2.305. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Rougeon F., Mach B. Cloning and amplification of rabbit alpha- and beta-globin gene sequences into Escherichia coli plasmids. J Biol Chem. 1977 Apr 10;252(7):2209–2217. [PubMed] [Google Scholar]
  32. Roychoudhury R., Jay E., Wu R. Terminal labeling and addition of homopolymer tracts to duplex DNA fragments by terminal deoxynucleotidyl transferase. Nucleic Acids Res. 1976 Jan;3(1):101–116. doi: 10.1093/nar/3.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Sakano H., Hüppi K., Heinrich G., Tonegawa S. Sequences at the somatic recombination sites of immunoglobulin light-chain genes. Nature. 1979 Jul 26;280(5720):288–294. doi: 10.1038/280288a0. [DOI] [PubMed] [Google Scholar]
  34. Sanger F., Nicklen S., Coulson A. R. DNA sequencing with chain-terminating inhibitors. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5463–5467. doi: 10.1073/pnas.74.12.5463. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schilling J., Clevinger B., Davie J. M., Hood L. Amino acid sequence of homogeneous antibodies to dextran and DNA rearrangements in heavy chain V-region gene segments. Nature. 1980 Jan 3;283(5742):35–40. doi: 10.1038/283035a0. [DOI] [PubMed] [Google Scholar]
  36. Seidman J. G., Max E. E., Leder P. A kappa-immunoglobulin gene is formed by site-specific recombination without further somatic mutation. Nature. 1979 Aug 2;280(5721):370–375. doi: 10.1038/280370a0. [DOI] [PubMed] [Google Scholar]
  37. Siekevitz M., Gefter M. L., Brodeur P., Riblet R., Marshak-Rothstein A. The genetic basis of antibody production: the dominant anti-arsonate idiotype response of the strain A mouse. Eur J Immunol. 1982 Dec;12(12):1023–1032. doi: 10.1002/eji.1830121208. [DOI] [PubMed] [Google Scholar]
  38. Sommé G., Serra J. R., Leclercq L., Moreau J. L., Mazie J. C., Moinier D., Fougerèau M., Thèze J. Contribution of the H- and L-chains and of the binding site to the idiotypic specificities of mouse anti-GAT antibodies. Mol Immunol. 1982 Aug;19(8):1011–1019. doi: 10.1016/0161-5890(82)90309-1. [DOI] [PubMed] [Google Scholar]
  39. 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]
  40. Thèze J., Moreau J. L. Genetic control of the immune response to the GAT terpolymer. I. Interstrain and interspecies cross-reactive idiotype. Ann Immunol (Paris) 1978 Jul-Sep;129 100(5):721–726. [PubMed] [Google Scholar]
  41. White-Scharf M. E., Imanishi-Kari T. Characterization of the NPa idiotype through the analysis of monoclonal BALB/c anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibodies. Eur J Immunol. 1981 Nov;11(11):897–904. doi: 10.1002/eji.1830111109. [DOI] [PubMed] [Google Scholar]

Articles from Nucleic Acids Research are provided here courtesy of Oxford University Press

RESOURCES