Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1985 Mar 1;161(3):547–562. doi: 10.1084/jem.161.3.547

Human immune response to group A streptococcal carbohydrate (A-CHO). I. Quantitative and qualitative analysis of the A-CHO-specific B cell population responding in vitro to polyclonal and specific activation

PMCID: PMC2187588  PMID: 2579184

Abstract

The immune response to the group-specific carbohydrate of group A streptococci (A-CHO) provides an informative in vitro model for the investigation of several aspects of human anticarbohydrate immune responses. A-CHO-specific B cells can be polyclonally activated by pokeweed mitogen (PWM), and, specifically, by in vitro immunization with streptococcal vaccine. High levels of A-CHO-specific antibodies, mainly directed to the immunodominant side chain N-acetyl-D-glucosamine (GlcNAc), occur in healthy adult individuals. Serum antibody levels are reflected in high frequencies of precursor B cells among peripheral blood lymphocytes. In one particular case, greater than 15% of all B cells activated by PWM for IgM production were found to produce IgM anti-A-CHO antibodies, as determined in limiting dilution experiments, as well as by analyzing Ig concentrations in bulk culture experiments. The case with the lowest proportion observed had 0.3% A-CHO-specific B cells among IgM-producing B cells. Preferential PWM activation of anti- A-CHO-producing B cells could be excluded. The comparison of the proportions of anti-A-CHO IgM produced in vivo, and of B cells producing antibodies of this specificity in peripheral blood, suggests a similar distribution of specific precursor B cells in the antibody- producing lymphoid tissue compartments and in peripheral blood. However, nearly all specific antibodies produced in vitro belong to the IgM isotype, whereas IgG anti-A-CHO in high amounts, mostly exceeding the specific IgM, was found only among anti-A-CHO antibodies produced in vivo. Low anti-A-CHO IgG production was seen in polyclonally activated as well as in antigen-activated cultures, whereas, in contrast, total IgG was produced in considerable amounts after polyclonal activation. This suggests a different distribution pattern, and/or diverse differentiation requirements for anti-A-CHO-producing B cells, compared with other B cell species.

Full Text

The Full Text of this article is available as a PDF (1,016.8 KB).

Selected References

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

  1. Ahmed A., Scher I., Sharrow S. O., Smith A. H., Paul W. E., Sachs D. H., Sell K. W. B-lymphocyte heterogeneity: development and characterization of an alloantiserum which distinguishes B-lymphocyte differentiation alloantigens. J Exp Med. 1977 Jan 1;145(1):101–110. doi: 10.1084/jem.145.1.101. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Andersson J., Coutinho A., Lernhardt W., Melchers F. Clonal growth and maturation to immunoglobulin secretion in vitro of every growth-inducible B lymphocyte. Cell. 1977 Jan;10(1):27–34. doi: 10.1016/0092-8674(77)90136-2. [DOI] [PubMed] [Google Scholar]
  3. Andersson J., Coutinho A., Melchers F. Frequencies of mitogen-reactive B cells in the mouse. II. Frequencies of B cells producing antibodies which lyse sheep or horse erythrocytes, and trinitrophenylated or nitroiodophenylated sheep erythrocytes. J Exp Med. 1977 Jun 1;145(6):1520–1530. doi: 10.1084/jem.145.6.1520. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Asano Y., Singer A., Hodes R. J. Role of the major histocompatibility complex in T cell activation of B cell subpopulations. Major histocompatibility complex-restricted and -unrestricted B cell responses are mediated by distinct B cell subpopulations. J Exp Med. 1981 Oct 1;154(4):1100–1115. doi: 10.1084/jem.154.4.1100. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Braun D. G., Eichmann K., Krause R. M. Rabbit antibodies to streptococcal carbohydrates. Influence of primary and secondary immunization and of possible genetic factors on the antibody response. J Exp Med. 1969 Apr 1;129(4):809–830. doi: 10.1084/jem.129.4.809. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Braun D. G., Jaton J. C. Homogeneous antibodies: induction and value as probe for the antibody problem. Curr Top Microbiol Immunol. 1974;66:29–76. doi: 10.1007/978-3-642-65908-9_2. [DOI] [PubMed] [Google Scholar]
  7. Braun D. G., Kindred B., Jacobson E. B. Streptococcal group A carbohydrate antibodies in mice: evidence for strain differences in magnitude and restriction of the response, and for thymus dependence. Eur J Immunol. 1972 Apr;2(2):138–143. doi: 10.1002/eji.1830020209. [DOI] [PubMed] [Google Scholar]
  8. Braun D. G., Quintáns J., Luzzati A. L., Lefkovits I., Read S. E. Antibody response of rabbit blood lymphocytes in vitro. Kinetics, clone size, and clonotype analysis in response to streptococcal group polysaccharide antigens. J Exp Med. 1976 Feb 1;143(2):360–371. doi: 10.1084/jem.143.2.360. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Click R. E., Benck L., Alter B. J. Immune responses in vitro. I. Culture conditions for antibody synthesis. Cell Immunol. 1972 Feb;3(2):264–276. doi: 10.1016/0008-8749(72)90165-7. [DOI] [PubMed] [Google Scholar]
  10. Coligan J. E., Schnute W. C., Jr, Kindt T. J. Immunochemical and chemical studies on streptococcal group-specific carbohydrates. J Immunol. 1975 Jun;114(6):1654–1658. [PubMed] [Google Scholar]
  11. Dosch H-M, Gelfand E. W. Generation of human plaque-forming cells in culture: tissue distribution, antigenic and cellular requirements. J Immunol. 1977 Jan;118(1):302–308. [PubMed] [Google Scholar]
  12. Eichmann K., Braun D. G., Krause R. M. Influence of genetic factors on the magnitude and the heterogeneity of the immune response in the rabbit. J Exp Med. 1971 Jul 1;134(1):48–65. doi: 10.1084/jem.134.1.48. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Eichmann K. Expression and function of idiotypes of lymphocytes. Adv Immunol. 1978;26:195–254. doi: 10.1016/s0065-2776(08)60231-x. [DOI] [PubMed] [Google Scholar]
  14. Eichmann K. Idiotype expression and the inheritance of mouse antibody clones. J Exp Med. 1973 Mar 1;137(3):603–621. doi: 10.1084/jem.137.3.603. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Eichmann K. Idiotypic identity of antibodies to streptococcal carbohydrate in inbred mice. Eur J Immunol. 1972 Aug;2(4):301–307. doi: 10.1002/eji.1830020402. [DOI] [PubMed] [Google Scholar]
  16. Emmrich F., Greger B., Eichmann K. A cross-reacting human idiotype (B17) associated with antibodies to N-acetyl-D-glucosamine. Specificity, immunoglobulin class association, and distribution in the population. Eur J Immunol. 1983 Apr;13(4):273–278. doi: 10.1002/eji.1830130402. [DOI] [PubMed] [Google Scholar]
  17. Geha R. S., Mudawwar F., Schneeberger E. The specificity of T-cell helper factor in man. J Exp Med. 1977 Jun 1;145(6):1436–1448. doi: 10.1084/jem.145.6.1436. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. KARAKAWA W. W., OSTERLAND C. K., KRAUSE R. DETECTION OF STREPTOCOCCAL GROUP-SPECIFIC ANTIBODIES IN HUMAN SERA. J Exp Med. 1965 Aug 1;122:195–205. doi: 10.1084/jem.122.2.195. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Krause R. M. The search for antibodies with molecular uniformity. Adv Immunol. 1970;12:1–56. doi: 10.1016/s0065-2776(08)60167-4. [DOI] [PubMed] [Google Scholar]
  20. Lane H. C., Volkman D. J., Whalen G., Fauci A. S. In vitro antigen-induced, antigen-specific antibody production in man. Specific and polyclonal components, kinetics, and cellular requirements. J Exp Med. 1981 Oct 1;154(4):1043–1057. doi: 10.1084/jem.154.4.1043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Martínez-Maza O., Britton S. Frequencies of the separate human B cell subsets activatable to Ig secretion by Epstein-Barr virus and pokeweed mitogen. J Exp Med. 1983 Jun 1;157(6):1808–1814. doi: 10.1084/jem.157.6.1808. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Pavlovskis O., Slade H. D. Adsorption of 3H-fatty acid esters of streptococcal groups A and E cell wall polysaccharide antigens by red blood cells and their effect on hemagglutination. J Bacteriol. 1969 Nov;100(2):641–646. doi: 10.1128/jb.100.2.641-646.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Perlmutter R. M., Nahm M., Stein K. E., Slack J., Zitron I., Paul W. E., Davie J. M. Immunoglobulin subclass-specific immunodeficiency in mice with an X-linked B-lymphocyte defect. J Exp Med. 1979 Apr 1;149(4):993–998. doi: 10.1084/jem.149.4.993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Read S. E., Braun D. G. In vitro antibody response of primed rabbit peripheral blood lymphocytes to group A variant streptococcal polysaccharide. Eur J Immunol. 1974 Jun;4(6):422–426. doi: 10.1002/eji.1830040607. [DOI] [PubMed] [Google Scholar]
  25. Riesen W. F., Skvaril F., Braun D. G. Natural infection of man with group A streptococci. Levels; restriction in class, subclass, and type; and clonal appearance of polysaccharide-group-specific antibodies. Scand J Immunol. 1976;5(4):383–390. doi: 10.1111/j.1365-3083.1976.tb00292.x. [DOI] [PubMed] [Google Scholar]
  26. SCHMIDT W. C., MOORE D. J. THE DETERMINATION OF ANTIBODY TO GROUP A STREPTOCOCCAL POLYSACCHARIDE IN HUMAN SERA BY HEMAGGLUTINATION. J Exp Med. 1965 May 1;121:793–806. doi: 10.1084/jem.121.5.793. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Stevens R. H., Macy E., Thiele C. J. Evidence that pokeweed-mitogen-reactive B cells are pre-committed in vivo to the high-rate secretion of a single immunoglobulin isotype in vitro. Scand J Immunol. 1981 Nov;14(5):449–457. doi: 10.1111/j.1365-3083.1981.tb00587.x. [DOI] [PubMed] [Google Scholar]
  28. Volkman D. J., Lane H. C., Fauci A. S. Antigen-induced in vitro antibody production in humans: a model for B cell activation and immunoregulation. Proc Natl Acad Sci U S A. 1981 Apr;78(4):2528–2531. doi: 10.1073/pnas.78.4.2528. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Yarchoan R., Tosato G., Blaese R. M., Simon R. M., Nelson D. L. Limiting dilution analysis of Epstein-Barr virus-induced immunoglobulin production by human B cells. J Exp Med. 1983 Jan 1;157(1):1–14. doi: 10.1084/jem.157.1.1. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Zenke G., Eichmann K., Emmrich F. Characterization of a major human antibody clonotype (1A) by monoclonal antibodies to combining site-associated idiotopes. Eur J Immunol. 1984 Feb;14(2):164–170. doi: 10.1002/eji.1830140211. [DOI] [PubMed] [Google Scholar]

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

RESOURCES