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. 1981 Feb 1;153(2):352–364. doi: 10.1084/jem.153.2.352

Anti-phosphocholine hybridoma antibodies. I. Direct evidence for three distinct families of antibodies in the murine response

JL Claflin, S Hudak, A Maddalena
PMCID: PMC2186073  PMID: 6787165

Abstract

Biochemical and serological studies were performed on more than 400 anti- phosphocholine (PC) hybridoma proteins (HP) derived from six strains of mice; 26 of these HP were examined in detail. All HP possessed specificity for PC, and all those tested contained an H-chain idiotypic determinant, V(H)-PC, which is shared by PC-binding myeloma proteins (BMP) and anti-PC antibodies. Among the HP, three well-defined and distinct families that correlated well with previous studies on serum anti-PC antibodies were identified. The largest group shared idotypic determinants, an L-chain isoelectric focusing (IEF) pattern, and a binding site specificity with the PC-BMP, T15. Using the same criteria, a second group was found to be strikingly similar to another PC-BMP, M603. The third group possessed an idiotypic determinant and an L-chain IEF profile similar to M511, but differences in binding site specificities were observed among the HP. The latter two groups contained members whose L-chain IEF profiles were not identical to other members of that group. Thus, among strains there is a remarkable degree of conservation among responding anti-PC antibodies, in both the kinds of anti-PC families that exist and the immunochemical and structural characteristics of various members within a family. Differences in at least one parameter were observed in each family, demonstrating that even a relatively restricted response is heterogeneous. However, this diversity seems to operate within certain constraints.

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Selected References

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  1. Capra J. D., Kehoe J. M. Structure of antibodies with shared idiotypy: the complete sequence of the heavy chain variable regions of two immunoglobulin M anti-gamma globulins. Proc Natl Acad Sci U S A. 1974 Oct;71(10):4032–4036. doi: 10.1073/pnas.71.10.4032. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Claflin J. L. Clonal nature of the immune response to phosphorylcholine. VIII. Evidence that antibodies bearing T15 idiotypic determinants in Ighj mice comprise a family of antibodies. J Immunol. 1980 Aug;125(2):559–563. [PubMed] [Google Scholar]
  3. Claflin J. L., Cubberley M. Clonal nature of the immune response to phosphocholine. VI. Molecular uniformity of a single idiotype among BALB/c mice. J Immunol. 1978 Oct;121(4):1410–1415. [PubMed] [Google Scholar]
  4. Claflin J. L., Cubberley M. Clonal nature of the immune response to phosphocholine. VII. Evidence throughout inbred mice for molecular similarities among antibodies bearing the T15 idiotype. J Immunol. 1980 Aug;125(2):551–558. [PubMed] [Google Scholar]
  5. Claflin J. L., Davie J. M. Clonal nature of the immune response to phosphorylcholine (PC). V. Cross-idiotypic specificity among heavy chains of murine anti-PC antibodies and PC-binding myeloma proteins. J Exp Med. 1975 May 1;141(5):1073–1083. doi: 10.1084/jem.141.5.1073. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Claflin J. L., Davie J. M. Clonal nature of the immune response to phosphorylcholine. IV. Idiotypic uniformity of binding site-associated antigenic determinants among mouse antiphosphorylcholine antibodies. J Exp Med. 1974 Sep 1;140(3):673–686. doi: 10.1084/jem.140.3.673. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Claflin J. L. Genetic marker in the variable region of kappa chains of mouse anti-phosphorylcholine antibodies. Eur J Immunol. 1976 Sep;6(9):666–668. doi: 10.1002/eji.1830060915. [DOI] [PubMed] [Google Scholar]
  8. Claflin J. L., Rudikoff S. Structural evidence for a polymorphic or allelic form of the heavy chain variable region. J Immunol. 1979 Apr;122(4):1402–1406. [PubMed] [Google Scholar]
  9. Claflin J. L. Uniformity in the clonal repertoire for the immune response to phosphorylcholine in mice. Eur J Immunol. 1976 Oct;6(10):669–674. doi: 10.1002/eji.1830061002. [DOI] [PubMed] [Google Scholar]
  10. Claflin J. L., Wei S. P. Control of V kappa expression in the mouse. I. Unexpected expression of the V kappa allele, Igk-Pcb, in a somatic cell hybrid of AKR (Igk-Pca) origin. J Immunol. 1979 Sep;123(3):1051–1056. [PubMed] [Google Scholar]
  11. Clevinger B., Schilling J., Hood L., Davie J. M. Structural correlates of cross-reactive and individual idiotypic determinants on murine antibodies to alpha-(1 leads to 3) dextran. J Exp Med. 1980 May 1;151(5):1059–1070. doi: 10.1084/jem.151.5.1059. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Feizi T., Kunkel H. G., Roelcke D. Cross idiotypic specificity among cold agglutinins in relation to combining activity for blood group-related antigens. Clin Exp Immunol. 1974 Oct;18(2):283–293. [PMC free article] [PubMed] [Google Scholar]
  13. Gearhart P. J., Sigal N. H., Klinman N. R. The monoclonal anti-phosphorylcholine antibody response in several murine strains: genetic implications of a diverse repertoire. J Exp Med. 1977 Apr 1;145(4):876–891. doi: 10.1084/jem.145.4.876. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Gefter M. L., Margulies D. H., Scharff M. D. A simple method for polyethylene glycol-promoted hybridization of mouse myeloma cells. Somatic Cell Genet. 1977 Mar;3(2):231–236. doi: 10.1007/BF01551818. [DOI] [PubMed] [Google Scholar]
  15. Jolley M. E., Glaudemans C. P., Rudikoff S., Potter M. Structural requirements for the binding of derivatives of D-galactose to two homogeneous murine immunoglobulins. Biochemistry. 1974 Jul 16;13(15):3179–3184. doi: 10.1021/bi00712a028. [DOI] [PubMed] [Google Scholar]
  16. Ju S. T., Pierres M., Germain R. N., Benacerraf B., Dorf M. E. Idiotypic analysis of anti-GAT antibodies. VI. Identification and strain distribution of the GA-1 idiotype. J Immunol. 1979 Dec;123(6):2505–2510. [PubMed] [Google Scholar]
  17. Keren D. F. Enzyme-linked immunosorbent assay for immunoglobulin G and immunoglobulin A antibodies to Shigella flexneri antigens. Infect Immun. 1979 May;24(2):441–448. doi: 10.1128/iai.24.2.441-448.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Kocher H. P., Berek C., Schreier M. H., Cosenza H., Jaton J. C. Phosphorylcholine-binding hybridoma proteins of normal and idiotypically suppressed BALB/c mice II. Variable region N-terminal amino acid sequences. Eur J Immunol. 1980 Apr;10(4):264–267. doi: 10.1002/eji.1830100408. [DOI] [PubMed] [Google Scholar]
  19. 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]
  20. Lamoyi E., Estess P., Capra J. D., Nisonoff A. Heterogeneity of an intrastrain cross-reactive idiotype associated with anti-p-azophenylarsonate antibodies of A/J mice. J Immunol. 1980 Jun;124(6):2834–2840. [PubMed] [Google Scholar]
  21. Lernhardt W., Andersson J., Coutinho A., Melchers F. Cloning of murine transformed cell lines in suspension culture with efficiencies near 100%. Exp Cell Res. 1978 Feb;111(2):309–316. doi: 10.1016/0014-4827(78)90175-1. [DOI] [PubMed] [Google Scholar]
  22. Marshak-Rothstein A., Siekevitz M., Margolies M. N., Mudgett-Hunter M., Gefter M. L. Hybridoma proteins expressing the predominant idiotype of the antiazophenylarsonate response of A/J mice. Proc Natl Acad Sci U S A. 1980 Feb;77(2):1120–1124. doi: 10.1073/pnas.77.2.1120. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Pierres M., Ju S. T., Waltenbaugh C., Dorf M. E., Benacerraf B., Germain R. N. Fine specificity of antibodies to poly(Glu60Ala30Tyr10) produced by hybrid cell lines. Proc Natl Acad Sci U S A. 1979 May;76(5):2425–2429. doi: 10.1073/pnas.76.5.2425. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Rudikoff S., Potter M. Allelic forms of the immunoglobulin heavy chain variable region. J Immunol. 1980 May;124(5):2089–2092. [PubMed] [Google Scholar]
  25. Rudikoff S., Rao D. N., Glaudemans C. P., Potter M. kappa Chain joining segments and structural diversity of antibody combining sites. Proc Natl Acad Sci U S A. 1980 Jul;77(7):4270–4274. doi: 10.1073/pnas.77.7.4270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Ruppert V. J., Williams K., Claflin J. L. Specific clonal regulation in the response to phosphocholine. I. Genetic analysis of the response of a distinct idiotype (M511 Id). J Immunol. 1980 Mar;124(3):1068–1074. [PubMed] [Google Scholar]
  27. Schroer K. R., Kim K. J., Prescott B., Baker P. J. Generation of anti-type III pneumococcal polysaccharide hybridomas from mice with an X-linked B-lymphocyte defect. J Exp Med. 1979 Sep 19;150(3):698–702. doi: 10.1084/jem.150.3.698. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Seidman J. G., Leder A., Edgell M. H., Polsky F., Tilghman S. M., Tiemeier D. C., Leder P. Multiple related immunoglobulin variable-region genes identified by cloning and sequence analysis. Proc Natl Acad Sci U S A. 1978 Aug;75(8):3881–3885. doi: 10.1073/pnas.75.8.3881. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Shulman M., Wilde C. D., Köhler G. A better cell line for making hybridomas secreting specific antibodies. Nature. 1978 Nov 16;276(5685):269–270. doi: 10.1038/276269a0. [DOI] [PubMed] [Google Scholar]
  30. Williams K. R., Claflin J. L. Clonotypes of anti-phosphocholine antibodies induced with Proteus morganii (Potter). I. Structural and idiotypic similarities in a diverse repertoire. J Immunol. 1980 Dec;125(6):2429–2436. [PubMed] [Google Scholar]
  31. Willims R. C., Jr, Kunkel H. G., Capra J. D. Antigenic specificities related to the cold agglutinin activity of gamma M globulins. Science. 1968 Jul 26;161(3839):379–381. doi: 10.1126/science.161.3839.379. [DOI] [PubMed] [Google Scholar]
  32. Wolfe J., Claflin J. L. Clonal nature of the immune response to phosphocholine. IX. Heterogeneity among antibodies bearing M511 idiotypic determinants. J Immunol. 1980 Dec;125(6):2397–2401. [PubMed] [Google Scholar]

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