Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1985 Jun 1;161(6):1272–1292. doi: 10.1084/jem.161.6.1272

Heavy chain variable region. Multiple gene segments encode anti-4- (hydroxy-3-nitro-phenyl)acetyl idiotypic antibodies

PMCID: PMC2187640  PMID: 3925064

Abstract

The hapten (4-hydroxy-3-nitrophenyl)acetyl (NP), when conjugated to carrier proteins, elicits a characteristic idiotypic response (NPb) in C57BL/6 mice. The response can be divided serologically into two distinct NPb-positive groups of antibodies. The first group consists of four crossreacting subgroups (I-IV), the second of two subgroups (V, VI). Some antibodies of subgroups I and II have been shown to express the unmutated heavy chain variable region (VH) germline gene 186.2. Antibodies of subgroups V and VI crossreact extensively with the NPa- positive antibodies of BALB/c mice. We sequenced heavy chain complementary DNA from eight hybridomas producing anti-NP antibodies. Six of these belong to subgroups V and VI, and two were NPa-positive hybridomas of BALB/c origin. All sequences were homologous to each other, and differed by approximately 80 basepairs from the 186.2 C57BL/6 germline VH gene. From our sequence and Southern blot analyses we suggest: (a) the NPb idiotypic response is the product of several VH germline genes, (b) some of these genes are very homologous to the gene coding for the BALB/c NPa idiotype, and might represent the C57BL/6 allelic forms of this gene, (c) the diversity regions of NPb and NPa- positive antibodies are diverse in length and amino acid composition, except for the first residue, which is always tyrosine, (d) all four heavy chain joining region gene segments are expressed without mutation. We discuss our data in terms of diversity in the germline VH gene repertoire, as well as diversity created by gene segment-joining events and somatic mutation.

Full Text

The Full Text of this article is available as a PDF (1.5 MB).

Selected References

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

  1. Alt F. W., Baltimore D. Joining of immunoglobulin heavy chain gene segments: implications from a chromosome with evidence of three D-JH fusions. Proc Natl Acad Sci U S A. 1982 Jul;79(13):4118–4122. doi: 10.1073/pnas.79.13.4118. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Azuma T., Igras V., Reilly E. B., Eisen H. N. Diversity at the variable-joining region boundary of lambda light chains has a pronounced effect on immunoglobulin ligand-binding activity. Proc Natl Acad Sci U S A. 1984 Oct;81(19):6139–6143. doi: 10.1073/pnas.81.19.6139. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. 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]
  4. Bothwell A. L., Paskind M., Reth M., Imanishi-Kari T., Rajewsky K., Baltimore D. Somatic variants of murine immunoglobulin lambda light chains. Nature. 1982 Jul 22;298(5872):380–382. doi: 10.1038/298380a0. [DOI] [PubMed] [Google Scholar]
  5. Cohen J. B., Givol D. Allelic immunoglobulin VH genes in two mouse strains: possible germline gene recombination. EMBO J. 1983;2(11):2013–2018. doi: 10.1002/j.1460-2075.1983.tb01693.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. 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]
  7. 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]
  8. Imanishi-Kari T., Rajnavölgyi E., Takemori T., Jack R. S., Rajewsky K. The effect of light chain gene expression on the inheritance of an idiotype associated with primary anti-(4-hydroxy-3-nitrophenyl)acetyl(NP) antibodies. Eur J Immunol. 1979 Apr;9(4):324–331. doi: 10.1002/eji.1830090414. [DOI] [PubMed] [Google Scholar]
  9. Imanishi T., Mäkelä O. Inheritance of antibody specificity. I. Anti-(4-hydroxy-3-nitrophenyl)acetyl of the mouse primary response. J Exp Med. 1974 Dec 1;140(6):1498–1510. doi: 10.1084/jem.140.6.1498. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Jack R. S., Imanishi-Kari T., Rajewsky K. Idiotypic analysis of the response of C57BL/6 mice to the (4-hydroxy-3-nitrophenyl)acetyl group. Eur J Immunol. 1977 Aug;7(8):559–565. doi: 10.1002/eji.1830070813. [DOI] [PubMed] [Google Scholar]
  11. Kaartinen M., Griffiths G. M., Hamlyn P. H., Markham A. F., Karjalainen K., Pelkonen J. L., Mäkelä O., Milstein C. Anti-oxazolone hybridomas and the structure of the oxazolone idiotype. J Immunol. 1983 Feb;130(2):937–945. [PubMed] [Google Scholar]
  12. Kaartinen M., Griffiths G. M., Markham A. F., Milstein C. mRNA sequences define an unusually restricted IgG response to 2-phenyloxazolone and its early diversification. 1983 Jul 28-Aug 3Nature. 304(5924):320–324. doi: 10.1038/304320a0. [DOI] [PubMed] [Google Scholar]
  13. Kabat E. A., Wu T. T., Bilofsky H. Evidence supporting somatic assembly of the DNA segments (minigenes), coding for the framework, and complementarity-determining segments of immunoglobulin variable regions. J Exp Med. 1979 Jun 1;149(6):1299–1313. doi: 10.1084/jem.149.6.1299. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Karjalainen K. Two major idiotypes in mouse anti-(4-hydroxy-3-nitro-phenyl)acetyl (NP) antibodies are controlled by "allelic" genes. Eur J Immunol. 1980 Feb;10(2):132–139. doi: 10.1002/eji.1830100212. [DOI] [PubMed] [Google Scholar]
  15. Kim S., Davis M., Sinn E., Patten P., Hood L. Antibody diversity: somatic hypermutation of rearranged VH genes. Cell. 1981 Dec;27(3 Pt 2):573–581. doi: 10.1016/0092-8674(81)90399-8. [DOI] [PubMed] [Google Scholar]
  16. Kunkel H. G., Agnello V., Joslin F. G., Winchester R. J., Capra J. D. Cross-idiotypic specificity among monoclonal IgM proteins with anti- -globulin activity. J Exp Med. 1973 Feb 1;137(2):331–342. doi: 10.1084/jem.137.2.331. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. 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]
  18. 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]
  19. 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]
  20. McKean D., Huppi K., Bell M., Staudt L., Gerhard W., Weigert M. Generation of antibody diversity in the immune response of BALB/c mice to influenza virus hemagglutinin. Proc Natl Acad Sci U S A. 1984 May;81(10):3180–3184. doi: 10.1073/pnas.81.10.3180. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Meek K., Jeske D., Slaoui M., Leo O., Urbain J., Capra J. D. Complete amino acid sequence of heavy chain variable regions derived from two monoclonal anti-p-azophenylarsonate antibodies of BALB/c mice expressing the major cross-reactive idiotype of the A/J strain. J Exp Med. 1984 Oct 1;160(4):1070–1086. doi: 10.1084/jem.160.4.1070. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Near R. I., Juszczak E. C., Huang S. Y., Sicari S. A., Margolies M. N., Gefter M. L. Expression and rearrangement of homologous immunoglobulin VH genes in two mouse strains. Proc Natl Acad Sci U S A. 1984 Apr;81(7):2167–2171. doi: 10.1073/pnas.81.7.2167. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Neuberger M. S., Calabi F. Reciprocal chromosome translocation between c-myc and immunoglobulin gamma 2b genes. Nature. 1983 Sep 15;305(5931):240–243. doi: 10.1038/305240a0. [DOI] [PubMed] [Google Scholar]
  24. Pech M., Höchtl J., Schnell H., Zachau H. G. Differences between germ-line and rearranged immunoglobulin V kappa coding sequences suggest a localized mutation mechanism. Nature. 1981 Jun 25;291(5817):668–670. doi: 10.1038/291668a0. [DOI] [PubMed] [Google Scholar]
  25. Perlmutter R. M., Klotz J. L., Bond M. W., Nahm M., Davie J. M., Hood L. Multiple VH gene segments encode murine antistreptococcal antibodies. J Exp Med. 1984 Jan 1;159(1):179–192. doi: 10.1084/jem.159.1.179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Pollok B. A., Kearney J. F., Vakil M., Perry R. P. A biological consequence of variation in the site of D-JH gene rearrangement. 1984 Sep 27-Oct 3Nature. 311(5984):376–379. doi: 10.1038/311376a0. [DOI] [PubMed] [Google Scholar]
  27. Reilly E. B., Reilly R. M., Breyer R. M., Sauer R. T., Eisen H. N. Amino acid and nucleotide sequences of variable regions of mouse immunoglobulin light chains of the lambda 3-subtype. J Immunol. 1984 Jul;133(1):471–475. [PubMed] [Google Scholar]
  28. Reth M., Hämmerling G. J., Rajewsky K. Analysis of the repertoire of anti-NP antibodies in C57BL/6 mice by cell fusion. I. Characterization of antibody families in the primary and hyperimmune response. Eur J Immunol. 1978 Jun;8(6):393–400. doi: 10.1002/eji.1830080605. [DOI] [PubMed] [Google Scholar]
  29. Reth M., Imanishi-Kari T., Rajewsky K. Analysis of the repertoire of anti-(4-hydroxy-3-nitrophenyl)acetyl (NP) antibodies in C 57 BL/6 mice by cell fusion. II. Characterization of idiotopes by monoclonal anti-idiotope antibodies. Eur J Immunol. 1979 Dec;9(12):1004–1013. doi: 10.1002/eji.1830091216. [DOI] [PubMed] [Google Scholar]
  30. Sakano H., Maki R., Kurosawa Y., Roeder W., Tonegawa S. Two types of somatic recombination are necessary for the generation of complete immunoglobulin heavy-chain genes. Nature. 1980 Aug 14;286(5774):676–683. doi: 10.1038/286676a0. [DOI] [PubMed] [Google Scholar]
  31. Siekevitz M., Huang S. Y., Gefter M. L. The genetic basis of antibody production: a single heavy chain variable region gene encodes all molecules bearing the dominant anti-arsonate idiotype in the strain A mouse. Eur J Immunol. 1983 Feb;13(2):123–132. doi: 10.1002/eji.1830130207. [DOI] [PubMed] [Google Scholar]
  32. Sims J., Rabbitts T. H., Estess P., Slaughter C., Tucker P. W., Capra J. D. Somatic mutation in genes for the variable portion of the immunoglobulin heavy chain. Science. 1982 Apr 16;216(4543):309–311. doi: 10.1126/science.6801765. [DOI] [PubMed] [Google Scholar]
  33. Slaughter C. A., Capra J. D. Amino acid sequence diversity within the family of antibodies bearing the major antiarsonate cross-reactive idiotype of the A strain mouse. J Exp Med. 1983 Nov 1;158(5):1615–1634. doi: 10.1084/jem.158.5.1615. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Tonegawa S. Somatic generation of antibody diversity. Nature. 1983 Apr 14;302(5909):575–581. doi: 10.1038/302575a0. [DOI] [PubMed] [Google Scholar]
  35. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Weigert M., Perry R., Kelley D., Hunkapiller T., Schilling J., Hood L. The joining of V and J gene segments creates antibody diversity. Nature. 1980 Jan 31;283(5746):497–499. doi: 10.1038/283497a0. [DOI] [PubMed] [Google Scholar]
  37. Weigert M., Raschke W. C., Carson D., Cohn M. Immunochemical analysis of the idiotypes of mouse myeloma proteins with specificity for levan or dextran. J Exp Med. 1974 Jan 1;139(1):137–147. doi: 10.1084/jem.139.1.137. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. 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]
  39. White-Scharf M. E., Imanishi-Kari T. Cross-reactivity of the NPa and NPb idiotypic responses of BALB/c and C57BL/6 mice to (4-hydroxy-3-nitrophenyl)acetyl (NP). Eur J Immunol. 1982 Nov;12(11):935–942. doi: 10.1002/eji.1830121108. [DOI] [PubMed] [Google Scholar]

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

RESOURCES