Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1972 Aug 1;136(2):241–260. doi: 10.1084/jem.136.2.241

THE MECHANISM OF ANTIGENIC STIMULATION OF PRIMARY AND SECONDARY CLONAL PRECURSOR CELLS

Norman R Klinman 1
PMCID: PMC2139200  PMID: 4114497

Abstract

Cell transfers to carrier-immunized irradiated mice have permitted an analysis of the in vitro stimulation of clonal precursors of anti-2,4-dinitrophenyl (DNP) antibody-producing cells derived from both immune and nonimmune mice. The results indicate that: (a) carrier-specific enhancement is obligatory for stimulation of primary precursor cells and increases both the size and number of detectable foci derived from secondary precursors. (b) This carrier-specific enhancement is most apparent in the stimulation of precursors of high-affinity antibody producer cells. (c) The antibody produced by primary foci, like that of secondary foci, appears homogeneous. (d) The frequency of clonal precursors in normal spleens is 38% that in spleens from mice 4–8 months after immunization, and the number of such precursors in normal spleens can be reduced fivefold by specific suppression of donor mice with soluble antigen. (e) The average of association constants of primary monofocal antibodies, like that of primary serum antibody produced in carrier-primed mice, is less than 10-fold lower than that of secondary clonal or serum antibody. (f) The affinity of primary monofocal antibodies shows a slight dependence on stimulating antigen concentration; however, a minimum threshold affinity consonant with stimulation is apparent. (g) Free hapten inhibits antigenic stimulation of primary precursor cells at a much lower concentration than is required for the inhibition of secondary precursors. These results are interpreted as indicating that (a) primary stimulation, like secondary stimulation, results from the selective stimulation by antigen of a population of cells differing from one another in their potential antibody product but each having only a single such product; (b) the antigen receptors of primary cells interact with antigen as if they are monovalent while receptors of secondary cells evidence multivalence; (c) antigenic stimulation appears to require both a relatively high affinity of receptors for bound antigen and an interlinking of receptors through such antigen; stimulation is thus seen as resulting from a stabilization of receptors within antigen-receptor aggregates to the cell surface; (d) T-cells appear to serve both in cross-linking antigens and in amplifying the size of stimulated clones.

Full Text

The Full Text of this article is available as a PDF (1.1 MB).

Selected References

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

  1. Ada G. L. Antigen binding cells in tolerance and immunity. Transplant Rev. 1970;5:105–129. doi: 10.1111/j.1600-065x.1970.tb00358.x. [DOI] [PubMed] [Google Scholar]
  2. Askonas B. A., Williamson A. R., Wright B. E. Selection of a single antibody-forming cell clone and its propagation in syngeneic mice. Proc Natl Acad Sci U S A. 1970 Nov;67(3):1398–1403. doi: 10.1073/pnas.67.3.1398. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bosma M., Weiler E. The clonal nature of antibody formation. I. Clones of antibody-forming cells of poly-D-alanine specificity. J Immunol. 1970 Jan;104(1):203–214. [PubMed] [Google Scholar]
  4. Braun D. G., Krause R. M. The individual antigenic specificity of antibodies to streptococcal carbohydrates. J Exp Med. 1968 Nov 1;128(5):969–989. doi: 10.1084/jem.128.5.969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Byers V. S., Sercarz E. E. The X-Y-Z scheme of immunocyte maturation. IV. The exhaustion of memory cells. J Exp Med. 1968 Feb 1;127(2):307–325. doi: 10.1084/jem.127.2.307. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Campbell P. A. Heterogeneity of antibodies produced by single hemolytic foci. Cell Immunol. 1971 Jun;2(3):250–258. doi: 10.1016/0008-8749(71)90044-x. [DOI] [PubMed] [Google Scholar]
  7. Cone R. E., Marchalonis J. J., Rolley R. T. Lymphocyte membrane dynamics. Metabolic release of cell surface proteins. J Exp Med. 1971 Dec 1;134(6):1373–1384. doi: 10.1084/jem.134.6.1373. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davie J. M., Paul W. E. Receptors on immunocompetent cells. IV. Direct measurement of avidity of cell receptors and cooperative binding of multivalent ligands. J Exp Med. 1972 Mar 1;135(3):643–659. doi: 10.1084/jem.135.3.643. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. EISEN H. N., SISKIND G. W. VARIATIONS IN AFFINITIES OF ANTIBODIES DURING THE IMMUNE RESPONSE. Biochemistry. 1964 Jul;3:996–1008. doi: 10.1021/bi00895a027. [DOI] [PubMed] [Google Scholar]
  10. GREENBURY C. L., MOORE D. H., NUNN L. A. THE REACTION WITH RED CELLS OF 7S RABBIT ANTIBODY, ITS SUB-UNITS AND THEIR RECOMBINANTS. Immunology. 1965 Apr;8:420–431. [PMC free article] [PubMed] [Google Scholar]
  11. Good A. H., Ceverha B. B. Immunologic assays for identifying single components in protein mixtures after isoelectric focusing in urea-containing acrylamide gels. J Immunol. 1971 Jun;106(6):1677–1680. [PubMed] [Google Scholar]
  12. Henry C., Kimura J., Wofsy L. Cell separation on affinity columns: the isolation of immunospecific precursor cells from unimmunized mice (lactoside hapten-lymphocyte receptors-immunology). Proc Natl Acad Sci U S A. 1972 Jan;69(1):34–36. doi: 10.1073/pnas.69.1.34. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Katz D. H., Paul W. E., Goidl E. A., Benacerraf B. Carrier function in anti-hapten antibody responses. 3. Stimulation of antibody synthesis and facilitation of hapten-specific secondary antibody responses by graft-versus-host reactions. J Exp Med. 1971 Feb 1;133(2):169–186. doi: 10.1084/jem.133.2.169. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Katz D. H., Paul W. E., Goidl E. A., Benacerraf B. Radioresistance of cooperative function of carrier-specific lymphocytes in antihapten antibody responses. Science. 1970 Oct 23;170(3956):462–464. doi: 10.1126/science.170.3956.462. [DOI] [PubMed] [Google Scholar]
  15. Kettman J., Dutton R. W. Radioresistance of the enhancing effect of cells from carrier-immunized mice in an in vitro primary immune response. Proc Natl Acad Sci U S A. 1971 Apr;68(4):699–703. doi: 10.1073/pnas.68.4.699. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Klinman N. R. Antibody with homogeneous antigen binding produced by splenic foci in organ culture. Immunochemistry. 1969 Sep;6(5):757–759. doi: 10.1016/0019-2791(67)90140-1. [DOI] [PubMed] [Google Scholar]
  17. Klinman N. R. Purification and analysis of "monofocal" antibody. J Immunol. 1971 May;106(5):1345–1352. [PubMed] [Google Scholar]
  18. 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]
  19. Klinman N. R., Rockey J. H., Frauenberger G., Karush F. Equine anti-hapten antibody. 3. The comparative properties of gamma G- and gammaA-antibodies. J Immunol. 1966 Apr;96(4):587–595. [PubMed] [Google Scholar]
  20. 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]
  21. 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]
  22. Kreth H. W., Williamson A. R. Cell surveillance model for lymphocyte cooperation. Nature. 1971 Dec 24;234(5330):454–456. doi: 10.1038/234454a0. [DOI] [PubMed] [Google Scholar]
  23. Luzzati A. L., Tosi R. M., Carbonara A. O. Electrophoretically homogeneous antibody synthesized by spleen foci of irradiated repopulated mice. J Exp Med. 1970 Aug 1;132(2):199–210. doi: 10.1084/jem.132.2.199. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Marchalonis J. J., Nossal G. J. Electrophoretic analysis of antibody produced by single cells. Proc Natl Acad Sci U S A. 1968 Nov;61(3):860–867. doi: 10.1073/pnas.61.3.860. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Miller J. F., Basten A., Sprent J., Cheers C. Interaction between lymphocytes in immune responses. Cell Immunol. 1971 Oct;2(5):469–495. doi: 10.1016/0008-8749(71)90057-8. [DOI] [PubMed] [Google Scholar]
  26. Mitchison N. A. The carrier effect in the secondary response to hapten-protein conjugates. II. Cellular cooperation. Eur J Immunol. 1971 Jan;1(1):18–27. doi: 10.1002/eji.1830010104. [DOI] [PubMed] [Google Scholar]
  27. Montgomery P. C., Rockey J. H., Williamson A. R. Homogeneous antibody elicited with dinitrophenyl-gramicidin-S (rabbit-isoelectric focusing). Proc Natl Acad Sci U S A. 1972 Jan;69(1):228–232. doi: 10.1073/pnas.69.1.228. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. OVARY Z., BENACERRAF B. IMMUNOLOGICAL SPECIFICITY OF THE SECONDARY RESPONSE WITH DINITROPHENYLATED PROTEINS. Proc Soc Exp Biol Med. 1963 Oct;114:72–76. doi: 10.3181/00379727-114-28589. [DOI] [PubMed] [Google Scholar]
  29. Paul W. E., Siskind G. W., Benacerraf B. Studies on the effect of the carrier molecule on antihapten antibody synthesis. II. Carrier specificity of anti-2,4-dinitrophenyl-poly-l-lysine antibodies. J Exp Med. 1966 Apr 1;123(4):689–705. doi: 10.1084/jem.123.4.689. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Pincus J. H., Jaton J. C., Bloch K. J., Haber E. Properties of structurally restricted antibody to type VIII pneumococcal polysaccharide. J Immunol. 1970 May;104(5):1149–1154. [PubMed] [Google Scholar]
  31. Playfair J. H. Response of mouse T and B lymphocytes to sheep erythrocytes. Nat New Biol. 1972 Jan 26;235(56):115–118. doi: 10.1038/newbio235115a0. [DOI] [PubMed] [Google Scholar]
  32. Raff M. C., Sternberg M., Taylor R. B. Immunoglobulin determinants on the surface of mouse lymphoid cells. Nature. 1970 Feb 7;225(5232):553–554. doi: 10.1038/225553a0. [DOI] [PubMed] [Google Scholar]
  33. Segal S., Globerson A., Feldman M., Haimovich J., Sela M. In vitro induction of a primary response to the dinitrophenyl determinant. J Exp Med. 1970 Jan 1;131(1):93–99. doi: 10.1084/jem.131.1.93. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Trump G. N., Singer S. J. Electrophoretically homogeneous anti-DNP antibodies with restricted isoelectric points elicited in mice by immunization with the antigen papain-S-DNPL. Proc Natl Acad Sci U S A. 1970 Jun;66(2):411–418. doi: 10.1073/pnas.66.2.411. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Vitetta E. S., Baur S., Uhr J. W. Cell surface immunoglobulin. II. Isolation and characterization of immunoglobulin from mouse splenic lymphocytes. J Exp Med. 1971 Jul 1;134(1):242–264. doi: 10.1084/jem.134.1.242. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Vitetta E. S., Uhr J. W. Release of cell surface immunoglobulin by mouse splenic lymphocytes. J Immunol. 1972 Feb;108(2):577–579. [PubMed] [Google Scholar]
  37. Zaretskaya Y. M., Panteleev E. I., Petrov R. V. Accumulation of antibody-forming cells in spleens of pre-immunized irradiated mice after transplantation of syngeneic bone marrow. Nature. 1969 Feb 8;221(5180):567–568. doi: 10.1038/221567a0. [DOI] [PubMed] [Google Scholar]

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

RESOURCES