Skip to main content
PMC home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1969 Nov 30;130(6):1367–1393. doi: 10.1084/jem.130.6.1367

LOCALIZATION OF IMMUNOLOGICAL COMPLEXES FIXING β1C (C3) IN GERMINAL CENTERS OF LYMPH NODES

K J Gajl-Peczalska 1, A J Fish 1, H J Meuwissen 1, D Frommel 1, R A Good 1
PMCID: PMC2138692  PMID: 4900926

Abstract

28 human and 60 experimentally stimulated rabbit lymph nodes were studied by means of light microscopy and immunofluorescence. 21 of the 28 human lymph nodes showed well-developed germinal centers. IgM, IgG, and the β1C component of complement were found in the same distribution within germinal centers when examined in serial cryostat sections. 36 rabbits were stimulated with Brucella antigen, and 24 rabbits with BSA. A strikingly consistent correlation between distribution and appearance of specific staining for rabbit β1C, IgM, and IgG was observed; when lymph nodes were stimulated with BSA, antigen and specific antibody were present. Treatment of unfixed sections with citrate-buffered saline at low pH resulted in complete elution of immunoglobulins, β1C, and BSA from rabbit germinal centers, and in marked diminution of IgG and IgM in human germinal centers, while at the same time plasma cells remained strongly fluorescent. Specific selective fixation of heterologous (human) complement in rabbit germinal centers positive for β1C, IgG, IgM, and BSA was also obtained. These data present strong evidence for the existence within germinal centers of antigen-antibody complexes which fix at least the β1C component of complement in vivo. The possibility of complete elution of immunoglobulins from rabbit germinal centers can be taken as evidence that, at least for 20 days after primary and secondary stimulation, a major component of the immunoglobulins present in germinal centers is not produced locally but accumulates at the surface of cells.

Full Text

The Full Text of this article is available as a PDF (1.6 MB).

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., AUSTIN C. M. ANTIGENS IN IMMUNITY. V. THE ABILITY OF CELLS IN LYMPHOID FOLLICLES TO RECOGNIZE FOREIGNNESS. Aust J Exp Biol Med Sci. 1964 Jun;42:331–346. [PubMed] [Google Scholar]
  2. ADA G. L., NOSSAL G. J., PYE J. ANTIGENS IN IMMUNITY. XI. THE UPTAKE OF ANTIGEN IN ANIMALS PREVIOUSLY RENDERED IMMUNOLOGICALLY TOLERANT. Aust J Exp Biol Med Sci. 1965 Jul;43:337–344. [PubMed] [Google Scholar]
  3. Asofsky R., Thorbecke G. J. SITES OF FORMATION OF IMMUNE GLOBULINS AND OF A COMPONENT OF C'(3): II. PRODUCTION OF IMMUNOELECTROPHORETICALLY IDENTIFIED SERUM PROTEINS BY HUMAN AND MONKEY TISSUES IN VITRO. J Exp Med. 1961 Sep 30;114(4):471–483. doi: 10.1084/jem.114.4.471. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bosman C., Feldman J. D., Pick E. Heterogeneity of antibody-forming cells. An electron microscopic analysis. J Exp Med. 1969 May 1;129(5):1029–1044. doi: 10.1084/jem.129.5.1029. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. CHAPLIN H., COHEN S., PRESS E. M. PREPARATION AND PROPERTIES OF THE PEPTIDE CHAINS OF NORMAL HUMAN 19 S GAMMA-GLOBULIN (IGM). Biochem J. 1965 Apr;95:256–261. doi: 10.1042/bj0950256. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. COONS A. H., LEDUC E. H., KAPLAN M. H. Localization of antigen in tissue cells. VI. The fate of injected foreign proteins in the mouse. J Exp Med. 1951 Feb;93(2):173–188. doi: 10.1084/jem.93.2.173. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. COTTIER H., ODARTCHENKO N., KEISER G., HESS M., STONER R. D. INCORPORATION OF TRITIATED NUCLEOSIDES AND AMINO ACIDS INTO LYMPHOID AND PLASMOCYTOID CELLS DURING SECONDARY RESPONSE TO TETANUS TOXOID IN MICE. Ann N Y Acad Sci. 1964 Feb 28;113:612–626. doi: 10.1111/j.1749-6632.1964.tb40694.x. [DOI] [PubMed] [Google Scholar]
  8. Cain W. A., Cooper M. D., Van Alten P. J., Good R. A. Development and function of the immunoglobulin-producing system. II. Role of the bursa in the development of humoral immunological competence. J Immunol. 1969 Mar;102(3):671–678. [PubMed] [Google Scholar]
  9. Cebra J. J., Goldstein G. Chromatographic purification of tetramethylrhodamine-immune globulin conjugates and their use in the cellular localization of rabbit gamma-globulin polypeptide chains. J Immunol. 1965 Aug;95(2):230–245. [PubMed] [Google Scholar]
  10. Cochrane C. G. The role of immune complexes and complement in tissue injury. J Allergy. 1968 Sep;42(3):113–129. doi: 10.1016/0021-8707(68)90085-3. [DOI] [PubMed] [Google Scholar]
  11. Cohen M. W., Jacobson E. B., Thorbecke G. J. Gamma-globulin and antibody formation in vitro. V. The secondary response made by splenic white and red pulp with reference to the role of secondary nodules. J Immunol. 1966 Jun;96(6):944–952. [PubMed] [Google Scholar]
  12. Cooper M. D., Cain W. A., Van Alten P. J., Good R. A. Development and function of the immunoglobulin producing system. I. Effect of bursectomy at different stages of development on germinal centers, plasma cells, immunoglobulins and antibody production. Int Arch Allergy Appl Immunol. 1969;35(3):242–252. [PubMed] [Google Scholar]
  13. FISHMAN M. Antibody formation in vitro. J Exp Med. 1961 Dec 1;114:837–856. doi: 10.1084/jem.114.6.837. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. FLIEDNER T., KESSE M., CRONKITE E. P., ROBERTSON J. S. CELL PROLIFERATION IN GERMINAL CENTERS OF THE RAT SPLEEN. Ann N Y Acad Sci. 1964 Feb 28;113:578–594. doi: 10.1111/j.1749-6632.1964.tb40692.x. [DOI] [PubMed] [Google Scholar]
  15. Glade P. R., Chessin L. N. Synthesis of B1c-B1a-globulin (C'3) by human lymphoid cells. Int Arch Allergy Appl Immunol. 1968;34(2):181–187. doi: 10.1159/000230108. [DOI] [PubMed] [Google Scholar]
  16. Good R. A., Finstad J., Cain W. A., Fish A., Perey D. Y., Gatti R. A. Models of immunologic diseases and disorders. Fed Proc. 1969 Jan-Feb;28(1):191–205. [PubMed] [Google Scholar]
  17. Good R. A., Quie P. G., Windhorst D. B., Page A. R., Rodey G. E., White J., Wolfson J. J., Holmes B. H. Fatal (chronic) granulomatous disease of childhood: a hereditary defect of leukocyte function. Semin Hematol. 1968 Jul;5(3):215–254. [PubMed] [Google Scholar]
  18. HANNA M. G., Jr AN AUTORADIOGRAPHIC STUDY OF THE GERMINAL CENTER IN SPLEEN WHITE PULP DURING EARLY INTERVALS OF THE IMMUNE RESPONSE. Lab Invest. 1964 Feb;13:95–104. [PubMed] [Google Scholar]
  19. HANNA M. G., Jr GERMINAL CENTER CHANGES AND PLASMA CELL REACTION DURING THE PRIMARY IMMUNE RESPONSE. Int Arch Allergy Appl Immunol. 1965;26:230–251. doi: 10.1159/000229574. [DOI] [PubMed] [Google Scholar]
  20. Hanna M. G., Jr, Congdon C. C., Wust C. J. Effect of antigen dose on lymphatic tissue germinal center changes. Proc Soc Exp Biol Med. 1966 Jan;121(1):286–290. doi: 10.3181/00379727-121-30760. [DOI] [PubMed] [Google Scholar]
  21. Hanna M. G., Jr, Swartzendruber D. C., Congdon C. C. Morphologic changes in spleen lymphatic tissue during antibody production. Exp Mol Pathol. 1966 Apr;(Suppl):75–87. [PubMed] [Google Scholar]
  22. Hochwald G. M., Thorbecke G. J., Asofsky R. SITES OF FORMATION OF IMMUNE GLOBULINS AND OF A COMPONENT OF C'(3): I. A NEW TECHNIQUE FOR THE DEMONSTRATION OF THE SYNTHESIS OF INDIVIDUAL SERUM PROTEINS BY TISSUES IN VITRO. J Exp Med. 1961 Sep 30;114(4):459–470. doi: 10.1084/jem.114.4.459. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. ISHIZAKA K. Gamma globulin and molecular mechanisms in hypersensitivity reactions. Prog Allergy. 1963;7:32–106. [PubMed] [Google Scholar]
  24. KAPLAN M. E., COONS A. H., DEANE H. W. Localization of antigen in tissue cells; cellular distribution of pneumococcal polysaccharides types II and III in the mouse. J Exp Med. 1950 Jan 1;91(1):15-30, 4 pl. doi: 10.1084/jem.91.1.15. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. LANGEVOORT H. L. The histophysiology of the antibody response. I. Histogenesis of the plasma cell reaction in rabbit spleen. Lab Invest. 1963 Jan;12:106–118. [PubMed] [Google Scholar]
  26. LEDUC E. H., COONS A. H., CONNOLLY J. M. Studies on antibody production. II. The primary and secondary responses in the popliteal lymph node of the rabbit. J Exp Med. 1955 Jul 1;102(1):61–72. doi: 10.1084/jem.102.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. LEDUC E. H., COONS A. H., CONNOLLY J. M. Studies on antibody production. II. The primary and secondary responses in the popliteal lymph node of the rabbit. J Exp Med. 1955 Jul 1;102(1):61–72. doi: 10.1084/jem.102.1.61. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. MARDINEY M. R., Jr, MUELLER-EBERHARD H. J. MOUSE BETA-1C-GLOBULIN: PRODUCTION OF ANTISERUM AND CHARACTERIZATION IN THE COMPLEMENT REACTION. J Immunol. 1965 Jun;94:877–882. [PubMed] [Google Scholar]
  29. MELLORS R. C., KORNGOLD L. THE CELLULAR ORIGIN OF HUMAN IMMUNOGLOBULINS (GAMMA-2, GAMMA-1M, GAMMA-1A). J Exp Med. 1963 Sep 1;118:387–396. doi: 10.1084/jem.118.3.387. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. MILLER J. J., 3rd, NOSSAL G. J. ANTIGENS IN IMMUNITY. VI. THE PHAGOCYTIC RETICULUM OF LYMPH NODE FOLLICLES. J Exp Med. 1964 Dec 1;120:1075–1086. doi: 10.1084/jem.120.6.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. MOVAT H. Z., FERNANDO N. V. THE FINE STRUCTURE OF THE LYMPHOID TISSUE DURING ANTIBODY FORMATION. Exp Mol Pathol. 1965 Apr;28:155–188. doi: 10.1016/0014-4800(65)90031-6. [DOI] [PubMed] [Google Scholar]
  32. McDevitt H. O., Askonas B. A., Humphrey J. H., Schechter I., Sela M. The localization of antigen in relation to specific antibody-producing cells. I. Use of a synthetic polypeptide [(T,G)-A--L] labelled with iodine-125. Immunology. 1966 Oct;11(4):337–351. [PMC free article] [PubMed] [Google Scholar]
  33. Michael A. F., Jr, Drummond K. N., Good R. A., Vernier R. L. Acute poststreptococcal glomerulonephritis: immune deposit disease. J Clin Invest. 1966 Feb;45(2):237–248. doi: 10.1172/JCI105336. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Mitchell J., Abbot A. Ultrastructure of the antigen-retaining reticulum of lymph node follicles as shown by high-resolution autoradiography. Nature. 1965 Oct 30;208(5009):500–502. doi: 10.1038/208500b0. [DOI] [PubMed] [Google Scholar]
  35. Müller-Eberhard H. J., Nilsson U. RELATION OF A beta(1)-GLYCOPROTEIN OF HUMAN SERUM TO THE COMPLEMENT SYSTEM. J Exp Med. 1960 Jan 31;111(2):217–234. doi: 10.1084/jem.111.2.217. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. NOSSAL G. J., ADA G. L., AUSTIN C. M. ANTIGENS IN IMMUNITY. IV. CELLULAR LOCALIZATION OF 125-I- AND 131-I-LABELLED FLAGELLA IN LYMPH NODES. Aust J Exp Biol Med Sci. 1964 Jun;42:311–330. [PubMed] [Google Scholar]
  37. Nilsson U. R., Müller-Eberhard H. J. Studies on the mode of action of the fifth, sixth and seventh component of human complement in immune haemolysis. Immunology. 1967 Jul;13(1):101–117. [PMC free article] [PubMed] [Google Scholar]
  38. ORTEGA L. G., MELLORS R. C. Cellular sites of formation of gamma globulin. J Exp Med. 1957 Nov 1;106(5):627–640. doi: 10.1084/jem.106.5.627. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. PORTER R. R. The hydrolysis of rabbit y-globulin and antibodies with crystalline papain. Biochem J. 1959 Sep;73:119–126. doi: 10.1042/bj0730119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. SIBOO R., VAS S. I. STUDIES ON IN VITRO ANTIBODY PRODUCTION. 3. PRODUCTION OF COMPLEMENT. Can J Microbiol. 1965 Jun;11:415–425. doi: 10.1139/m65-055. [DOI] [PubMed] [Google Scholar]
  41. Small P. A., Jr, Lamm M. E. Polypeptide chain structure of rabbit immunoglobulins. I. gamma-G-immunoglobulin. Biochemistry. 1966 Jan;5(1):259–267. doi: 10.1021/bi00865a034. [DOI] [PubMed] [Google Scholar]
  42. Stecher V. J., Morse J. H., Thorbecke G. J. Sites of production of primate serum proteins associated with complement system. Proc Soc Exp Biol Med. 1967 Feb;124(2):433–438. doi: 10.3181/00379727-124-31758. [DOI] [PubMed] [Google Scholar]
  43. Stecher V. J., Thorbecke G. J. Beta-1C and immune globulin formation in vitro by tissues from germ-free and conventional rodents of various ages. Immunology. 1967 Apr;12(4):475–487. [PMC free article] [PubMed] [Google Scholar]
  44. Stecher V. J., Thorbecke G. J. Sites of synthesis of serum proteins. 3. Production of beta-1-C, beta-1-E and transferrin by primate and rodent cell lines. J Immunol. 1967 Oct;99(4):660–668. [PubMed] [Google Scholar]
  45. Stecher V. J., Thorbecke G. J. Sites of synthesis of serum proteins. II. Medium requirements for serum protein production by rat macrophages. J Immunol. 1967 Oct;99(4):653–659. [PubMed] [Google Scholar]
  46. Stecher V. J., Thorbecke G. J. Sites of synthesis of serum proteins. OI. Serum proteins produced by macrophages in vitro. J Immunol. 1967 Oct;99(4):643–652. [PubMed] [Google Scholar]
  47. Thorbecke G. J., Hurlimann J. Les nodules lymphoïdes secondaires. Nouv Rev Fr Hematol. 1965 May-Jun;5(3):385–392. [PubMed] [Google Scholar]
  48. WARD P. A., COCHRANE C. G., MUELLER-EBERHARD H. J. THE ROLE OF SERUM COMPLEMENT IN CHEMOTAXIS OF LEUKOCYTES IN VITRO. J Exp Med. 1965 Aug 1;122:327–346. doi: 10.1084/jem.122.2.327. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Wakefield J. D., Thorbecke G. J. Relationship of germinal centers in lymphoid tissue to immunological memory. II. The detection of primed cells and their proliferation upon cell transfer to lethally irradiated syngeneic mice. J Exp Med. 1968 Jul 1;128(1):171–187. doi: 10.1084/jem.128.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  50. West C., Davis N. C., Forristal J., Herbst J., Spitzer R. Antigenic determinants of human beta-1c and beta-1g-globulins. J Immunol. 1966 Apr;96(4):650–658. [PubMed] [Google Scholar]
  51. Wood B. T., Thompson S. H., Goldstein G. Fluorescent antibody staining. 3. Preparation of fluorescein-isothiocyanate-labeled antibodies. J Immunol. 1965 Aug;95(2):225–229. [PubMed] [Google Scholar]

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

RESOURCES