Abstract
Immune responsiveness to phosphorylcholine (PC) in BALB/c mice has been characterized by combining (a) usuage of highly sensitive radioimmunoassays for quantitation of antibody, heavy-chain class, and idiotype on a weight basis; (b) isolation of PC-specific B cells in fragment cultures; and (c) stimulation in a carrier-primed environment with the PC hapten coupled to carrier through a tripeptide spacer in order to maximize carrier recognition. The specificity and accuracy of the radioimmunoassays have veen verified by specific inhibition, lack of nonspecific binding, and excellent concordance of values for monoclonal antibody concentration obtained independently for Fab and idiotype content. The latter evidence also serves as strong confirmation of the monoclonality of in vitro monofocal responses as well as the preservation of the idiotype on antibodies of differing immunoglobulin classes. The results indicate that while B cells expressing the TEPC 15 idiotype predominate, other idiotypes may be represented by 2-50% of PC-specific precursors, and monoclonal antibodies even of the TEPC 15 idiotype are produced in both the IgM and IgG1 immunoglobulin classes. These findings are confirmed by the analysis of serum antibodies produced in carrier-primed mice immunized with hapten coupled through a tripeptide spacer, thus re-emphasizint the enhancement of primary responsiveness, particularly IgG1 production, by maximizing carrier recognition. The finding of idiotype diversity in the PC response, as well as diversity of expression in terms of quantity and immunoglobulin class of antibody synthesized by the clonal progeny of B cells within the TEPC 15 clonotype, emphasize the heterogeneity of the B-cell population both in terms of specificity repertoire and the physiological state of cells even within a single clonotype.
Full Text
The Full Text of this article is available as a PDF (1.0 MB).
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Askenase P. W., Leonard E. J. Solid phase radioimmunoassay of human beta 1C globulin. Immunochemistry. 1970 Jan;7(1):29–41. doi: 10.1016/0019-2791(70)90028-5. [DOI] [PubMed] [Google Scholar]
- Askonas B. A., Williamson A. R. Dominance of a cell clone forming antibody to DNP. Nature. 1972 Aug 11;238(5363):339–341. doi: 10.1038/238339a0. [DOI] [PubMed] [Google Scholar]
- Carson D., Weigert M. Immunochemical analysis of the cross-reacting idiotypes of mouse myeloma proteins with anti-dextran activity and normal anti-dextran antibody. Proc Natl Acad Sci U S A. 1973 Jan;70(1):235–239. doi: 10.1073/pnas.70.1.235. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Chesebro B., Metzger H. Affinity labeling of a phosphorylcholine binding mouse myeloma protein. Biochemistry. 1972 Feb 29;11(5):766–771. doi: 10.1021/bi00755a014. [DOI] [PubMed] [Google Scholar]
- Claflin J. L., Lieberman R., Davie J. M. Clonal nature of the immune response to phosphorylcholine. I. Specificity, class, and idiotype of phosphorylcholine-binding receptors on lymphoid cells. J Exp Med. 1974 Jan 1;139(1):58–73. doi: 10.1084/jem.139.1.58. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Claflin J. L., Lieberman R., Davie J. M. Clonal nature of the immune response to phosphorylcholine. II. Idiotypic specificity and binding characteristics of anti-phosphorylcholine antibodies. J Immunol. 1974 May;112(5):1747–1756. [PubMed] [Google Scholar]
- Cosenza H., Köhler H. Specific inhibition of plaque formation to phosphorylcholine by antibody against antibody. Science. 1972 Jun 2;176(4038):1027–1029. doi: 10.1126/science.176.4038.1027. [DOI] [PubMed] [Google Scholar]
- Cosenza H., Köhler H. Specific suppression of the antibody response by antibodies to receptors. Proc Natl Acad Sci U S A. 1972 Sep;69(9):2701–2705. doi: 10.1073/pnas.69.9.2701. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Cuatrecasas P. Protein purification by affinity chromatography. Derivatizations of agarose and polyacrylamide beads. J Biol Chem. 1970 Jun;245(12):3059–3065. [PubMed] [Google Scholar]
- Goetzl E. J., Metzger H. Affinity labeling of a mouse myeloma protein which binds nitrophenyl ligands. Kinetics of labeling and isolation of a labeled peptide. Biochemistry. 1970 Mar 3;9(5):1267–1278. doi: 10.1021/bi00807a031. [DOI] [PubMed] [Google Scholar]
- Hood L., McKean D., Farnsworth V., Potter M. Mouse immunoglobulin chains. A survey of the amino-terminal sequences of kappa chains. Biochemistry. 1973 Feb;12(4):741–749. doi: 10.1021/bi00728a026. [DOI] [PubMed] [Google Scholar]
- Klinman N. R., Aschinazi G. The stimulation of splenic foci in vitro. J Immunol. 1971 May;106(5):1338–1344. [PubMed] [Google Scholar]
- Klinman N. R. Purification and analysis of "monofocal" antibody. J Immunol. 1971 May;106(5):1345–1352. [PubMed] [Google Scholar]
- Klinman N. R. Purification and analysis of "monofocal" antibody. J Immunol. 1971 May;106(5):1345–1352. [PubMed] [Google Scholar]
- Klinman N. R. Regain of homogeneous binding activity after recombination of chains of "mono- focal" antibody. J Immunol. 1971 May;106(5):1330–1337. [PubMed] [Google Scholar]
- Klinman N. R., Taylor R. B. General methods for the study of cells and serum during the immune response: the response to dinitrophenyl in mice. Clin Exp Immunol. 1969 Apr;4(4):473–487. [PMC free article] [PubMed] [Google Scholar]
- Klinman N. R. The mechanism of antigenic stimulation of primary and secondary clonal precursor cells. J Exp Med. 1972 Aug 1;136(2):241–260. doi: 10.1084/jem.136.2.241. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Klinman N. R. The secondary immune response to a hapten in vitro. Antigen concentration and the carrier effect. J Exp Med. 1971 May 1;133(5):963–972. doi: 10.1084/jem.133.5.963. [DOI] [PMC free article] [PubMed] [Google Scholar]
- LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
- Lieberman R., Humphrey W., Jr Association of H-2 types with genetic control of immune responsiveness to IgA allotypes in the mouse. Proc Natl Acad Sci U S A. 1971 Oct;68(10):2510–2513. doi: 10.1073/pnas.68.10.2510. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Lieberman R., Potter M., Mushinski E. B., Humphrey W., Jr, Rudikoff S. Genetics of a new IgVH (T15 idiotype) marker in the mouse regulating natural antibody to phosphorylcholine. J Exp Med. 1974 Apr 1;139(4):983–1001. doi: 10.1084/jem.139.4.983. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Mitchell G. F., Grumet F. C., McDevitt H. O. Genetic control of the immune response. The effect of thymectomy on the primary and secondary antibody response of mice to poly-L(tyr, glu)-poly-D, L-ala--poly-L-lys. J Exp Med. 1972 Jan;135(1):126–135. doi: 10.1084/jem.135.1.126. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Potter M., Lieberman R. Common individual antigenic determinants in five of eight BALB-c IgA myeloma proteins that bind phosphoryl choline. J Exp Med. 1970 Oct 1;132(4):737–751. doi: 10.1084/jem.132.4.737. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Press J. L., Klinman N. R. Frequency of hapten-specific B cells in neonatal and adult murine spleens. Eur J Immunol. 1974 Mar;4(3):155–159. doi: 10.1002/eji.1830040302. [DOI] [PubMed] [Google Scholar]
- Press J. L., Klinman N. R. Isoelectric analysis of neonatal monofocal antibody. Immunochemistry. 1973 Sep;10(9):621–627. doi: 10.1016/0019-2791(73)90164-x. [DOI] [PubMed] [Google Scholar]
- Press J. L., Klinman N. R. Monoclonal production of both IgM and IgG1 antihapten antibody. J Exp Med. 1973 Jul 1;138(1):300–305. doi: 10.1084/jem.138.1.300. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Robbins J. B., Haimovich J., Sela M. Purification of antibodies with immunoadsorbents prepared using bromoacetyl cellulose. Immunochemistry. 1967 Jan;4(1):11–22. doi: 10.1016/0019-2791(67)90192-9. [DOI] [PubMed] [Google Scholar]
- Sher A., Cohn M. Inheritance of an idiotype associated with the immune response of inbred mice to phosphorylcholine. Eur J Immunol. 1972 Aug;2(4):319–326. doi: 10.1002/eji.1830020405. [DOI] [PubMed] [Google Scholar]