Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1974 Jun 1;139(6):1582–1598. doi: 10.1084/jem.139.6.1582

EFFECTOR CELL BLOCKADE

A NEW MECHANISM OF IMMUNE HYPOREACTIVITY INDUCED BY MULTIVALENT ANTIGENS

John W Schrader 1, G J V Nossal 1
PMCID: PMC2139685  PMID: 4133616

Abstract

This study describes the effects of incubating antibody-forming cells (AFC), either as mass cell suspensions, or as single AFC in microdroplets, with antigens against which the cells display specificity. Most of the work was done with hapten-specific anti-DNP-AFC, but AFC with specificity against flagellar antigens or fowl gamma globulin (FGG) were also included. It was noted that 30-min incubation of AFC with highly multivalent forms of antigen caused a substantial partial suppression of the antibody-forming performance of the AFC as measured by a hemolytic plaque test. Thus, when cell suspensions containing anti-DNP plaque-forming cells (PFC), were incubated for 30 min at 37°C with 100 µg of DNP-polymerized flagellin (DNP-POL), the number of plaques appearing after washing of the cells and placing them in plaque-revealing erythrocyte monolayers was reduced to 50% or less compared with the number of plaques observed with control portions preincubated with medium alone. Preincubation with DNP-lysine, with oligovalent DNP-protein conjugates, or with irrelevant antigens produced no such inhibition. Studies where preinhibited PFC suspensions were mixed with control suspensions before assay showed that a nonspecific carryover of antigen into the assay system was not involved. The inhibitory effect could also be initiated by holding cells at 0°C with DNP-POL, but in that case, inhibition only became manifest after cells were incubated for 30 min at 37°C before being placed in plaque-revealing monolayers. This suggested that inhibition was initiated by adsorption of multivalent antigen onto PFC-surface Ig, but required some active process before secretion actually slowed down. The effect was dose- and time-dependent, antigen-specific, and generalized for all antigens studied. As well as yielding reduced plaque numbers, the preinhibited cells also gave smaller, more turbid plaques, suggesting a reduction in antibody-forming rate by each PFC rather than the elimination of PFC. Consistent with this suggestion was the observation that the degree of inhibition of plaque formation could be increased by decreasing the sensitivity of the assay so that only AFC secreting at high rates were detected. A micromanipulation study, where single PFC were subjected to inhibition, and were then tested for the rate at which they could cause hemolysis, showed a 68% inhibition of mean secretory rate. Micromanipulation studies were performed to test the amount of cell surface-associated Ig on control and preinhibited PFC. For this, single PFC were held with [125I]antiglobulin and quantitative radioautography was performed. No significant difference emerged, suggesting that retention of secreted Ig on cell-attached antigen was not the cause of inhibition. The results are discussed in the framework of tolerance models and blocking effects at the T-cell level by antigen-antibody complexes. The name effector cell blockade is suggested in the belief that the phenomenon may be a general one applying to both T and B cells.

Full Text

The Full Text of this article is available as a PDF (921.0 KB).

Selected References

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

  1. ADA G. L., NOSSAL G. J., PYE J., ABBOT A. ANTIGENS IN IMMUNITY. I. PREPARATION AND PROPERTIES OF FLAGELLAR ANTIGENS FROM SALMONELLA ADELAIDE. Aust J Exp Biol Med Sci. 1964 Jun;42:267–282. [PubMed] [Google Scholar]
  2. Andersson B. Studies on the regulation of avidity at the level of the single antibody-forming cell. The effect of antigen dose and time after immunization. J Exp Med. 1970 Jul 1;132(1):77–88. doi: 10.1084/jem.132.1.77. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Baker P. J., Stashak P. W., Amsbaugh D. F., Prescott B. Characterization of the antibody response to type 3 pneumococcal polysaccharide at the cellular level. II. Studies on the relative rate of antibody synthesis and release by antibody-producing cells. Immunology. 1971 Apr;20(4):481–492. [PMC free article] [PubMed] [Google Scholar]
  4. Britton S. Regulation of antibody synthesis against Escherichia coli endotoxin. IV. Induction of paralysis in vitro by treating normal lymphoid cells with antigen. J Exp Med. 1969 Mar 1;129(3):469–482. doi: 10.1084/jem.129.3.469. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CLAMAN H. N. TOLERANCE TO A PROTEIN ANTIGEN IN ADULT MICE AND THE EFFECT OF NONSPECIFIC FACTORS. J Immunol. 1963 Dec;91:833–839. [PubMed] [Google Scholar]
  6. Cunningham A. J., Szenberg A. Further improvements in the plaque technique for detecting single antibody-forming cells. Immunology. 1968 Apr;14(4):599–600. [PMC free article] [PubMed] [Google Scholar]
  7. Diener E., Feldmann M. Relationship between antigen and antibody-induced suppression of immunity. Transplant Rev. 1972;8:76–103. doi: 10.1111/j.1600-065x.1972.tb01565.x. [DOI] [PubMed] [Google Scholar]
  8. Feldman M. Induction fo B cell tolerance by antigen specific T cell factor. Nat New Biol. 1973 Mar 21;242(116):82–84. [PubMed] [Google Scholar]
  9. Feldmann M. Induction of immunity and tolerance in vitro by hapten protein conjugates. I. The relationship between the degree of hapten conjugation and the immunogenicity of dinitrophenylated polymerized flagellin. J Exp Med. 1972 Apr 1;135(4):735–753. doi: 10.1084/jem.135.4.735. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Feldmann M., Nossal G. J. Tolerance, enhancement and the regulation of interactions between T cells, B cells and macrophages. Transplant Rev. 1972;13:3–34. doi: 10.1111/j.1600-065x.1972.tb00058.x. [DOI] [PubMed] [Google Scholar]
  11. Hiramoto R. N., McGhee J. R., Hamlin N. M. Measurement of antibody release from single cells. I. J Immunol. 1972 Nov;109(5):961–967. [PubMed] [Google Scholar]
  12. Howard J. G. Cellular events in the induction and loss of tolerance to pneumococcal polysaccharides. Transplant Rev. 1972;8:50–75. doi: 10.1111/j.1600-065x.1972.tb01564.x. [DOI] [PubMed] [Google Scholar]
  13. Howard J. G., Elson J., Christie G. H., Kinsky R. G. Studies on immunological paralysis. II. The detection and significance of antibod-forming cells in the spleen during immunological paralysis with type 3 pneumococcal polysaccharide. Clin Exp Immunol. 1969 Jan;4(1):41–53. [PMC free article] [PubMed] [Google Scholar]
  14. Katz D. H., Davie J. M., Paul W. E., Benacerraf B. Carrier function in anti-hapten antibody responses. IV. Experimental conditions for the induction of hapten-specific tolerance or for the stimulation of anti-hapten anamnestic responses by "nonimmunogenic" hapten-polypeptide conjugates. J Exp Med. 1971 Jul 1;134(1):201–223. doi: 10.1084/jem.134.1.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kotlarski I., Courtenay B. M., Howard J. G. Studies on immunological paralysis. XI. A comparison of the tolerogenicity in vitro of levan and type 3 pneumococcal polysaccharide. Eur J Immunol. 1973 Aug;3(8):496–502. doi: 10.1002/eji.1830030809. [DOI] [PubMed] [Google Scholar]
  16. Langman R. E. The use of erythrocytes sensitized with flagellar antigens from Salmonella for the assay of antibody and antibody-forming cells. J Immunol Methods. 1972 Nov;2(1):59–66. doi: 10.1016/0022-1759(72)90018-x. [DOI] [PubMed] [Google Scholar]
  17. Miller J. F., Warner N. L. The immune response of normal, irradiated and thymectomized mice to fowl immunoglobulin G as detected by a hemolytic plaque technique. Int Arch Allergy Appl Immunol. 1971;40(1):59–71. doi: 10.1159/000230395. [DOI] [PubMed] [Google Scholar]
  18. Miranda J. J., Zola H., Howard J. G. Studies on immunological paralysis. X. Cellular characteristics of the induction and loss of tolerance to levan (Polyfructose). Immunology. 1972 Dec;23(6):843–855. [PMC free article] [PubMed] [Google Scholar]
  19. Mitchell G. F., Humphrey J. H., Wiliamson A. R. Inhibition of secondary anti-hapten responses with the hapten conjugated to type 3 pneumococcal polysaccharide. Eur J Immunol. 1972 Oct;2(5):460–467. doi: 10.1002/eji.1830020516. [DOI] [PubMed] [Google Scholar]
  20. Nossal G. J., Cunningham A., Mitchell G. F., Miller J. F. Cell to cell interaction in the immune response. 3. Chromosomal marker analysis of single antibody-forming cells in reconstituted, irradiated, or thymectomized mice. J Exp Med. 1968 Oct 1;128(4):839–853. doi: 10.1084/jem.128.4.839. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Nossal G. J., Lewis H. Variation in accessible cell surface immunoglobulin among antibody-firming cells. J Exp Med. 1972 Jun 1;135(6):1416–1422. doi: 10.1084/jem.135.6.1416. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Nossal G. J., Pike B. L., Katz D. H. Induction of B cell tolerance in vitro to 2,4-dinitrophenyl coupled to a copolymer of D-glutamic acid and D-lysine (DNP-D-GL). J Exp Med. 1973 Jul 1;138(1):312–317. doi: 10.1084/jem.138.1.312. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nossal G. J., Warner N. L., Lewis H., Sprent J. Quantitative features of a sandwich radioimmunolabeling technique for lymphocyte surface receptors. J Exp Med. 1972 Feb 1;135(2):405–428. doi: 10.1084/jem.135.2.405. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Schrader J. W. Induction of immunological tolerance to a thymus-dependent antigen in the absence of thymus-derived cells. J Exp Med. 1974 May 1;139(5):1303–1316. doi: 10.1084/jem.139.5.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Sjöberg O. Rapid breaking of tolerance against Escherichia coli lipopolysaccharide in vivo and in vitro. J Exp Med. 1972 Apr 1;135(4):850–859. doi: 10.1084/jem.135.4.850. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Sjögren H. O., Hellström I., Bansal S. C., Hellström K. E. Suggestive evidence that the "blocking antibodies" of tumor-bearing individuals may be antigen--antibody complexes. Proc Natl Acad Sci U S A. 1971 Jun;68(6):1372–1375. doi: 10.1073/pnas.68.6.1372. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Taylor R. B., Wortis H. H. Thymus dependence of antibody response: variation with dose of antigen and class of antibody. Nature. 1968 Nov 30;220(5170):927–928. doi: 10.1038/220927a0. [DOI] [PubMed] [Google Scholar]
  28. Weigle W. O. Immunological unresponsiveness. Adv Immunol. 1973;16:61–122. doi: 10.1016/s0065-2776(08)60296-5. [DOI] [PubMed] [Google Scholar]

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

RESOURCES