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. 1977 Feb;15(2):360–369. doi: 10.1128/iai.15.2.360-369.1977

Immunoglobulin and specific-antibody synthesis in vitro by enteral and nonenteral lymphoid tissues after subcutaneous cholera immunization.

A M Svennerholm, J Holmgren
PMCID: PMC421375  PMID: 844901

Abstract

An in vitro culture technique with synthesis of 14C-labeled protein has been used to study immunoglobulin and specific-antibody formation by spleen, mesenteric lymph nodes, Peyer's patches, and small intestine of rabbits, which were immunized twice subcutaneously with Vibrio cholerae lipopolysaccharide (LPS) and enterotoxin; saline-injected rabbits served as controls. Newly synthesized immunoglobulin G (IgG), IgA, and IgM were quantitated by liquid scintillation after their isolation by means of affinity chromatography from columns with immunoglobulin class-specific antibodies coupled to Sepharose. Specific antibodies were similarly measured after purification from gels derivatized with LPS or cholera toxin. The isolated antibodies had full biological activity as studied in protection tests. The immunization increased the overall IgM synthesis in the spleen. It also enhanced the production of IgA and IgG in Peyer's patches and of IgA in intestine. Significant synthesis of radiolabeled antibodies against both V. cholerae LPS and enterotoxin was found in spleen as well as in Peyer's patches of immunized animals. Titration with an enzyme-linked immunosorbent assay (ELISA) showed significant levels of IgG as well as IgA antibodies in incubation medium from all the studied tissues, whereas specific IgM was only found for spleen and mesenteric lymph nodes. Simultaneous tissue incubations at 37 degrees C and in an ice bath indicated that the major part of the antibodies registered with the ELISA represented de novo synthesis.

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Selected References

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  1. Axén R., Porath J., Ernback S. Chemical coupling of peptides and proteins to polysaccharides by means of cyanogen halides. Nature. 1967 Jun 24;214(5095):1302–1304. doi: 10.1038/2141302a0. [DOI] [PubMed] [Google Scholar]
  2. Crabbé P. A., Nash D. R., Bazin H., Eyssen H., Heremans J. F. Immunohistochemical observations on lymphoid tissues from conventional and germ-free mice. Lab Invest. 1970 May;22(5):448–457. [PubMed] [Google Scholar]
  3. Craig J. P. A permeability factor (toxin) found in cholera stools and culture filtrates and its neutralization by convalescent cholera sera. Nature. 1965 Aug 7;207(997):614–616. doi: 10.1038/207614a0. [DOI] [PubMed] [Google Scholar]
  4. Craig S. W., Cebra J. J. Peyer's patches: an enriched source of precursors for IgA-producing immunocytes in the rabbit. J Exp Med. 1971 Jul 1;134(1):188–200. doi: 10.1084/jem.134.1.188. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Craig S. W., Cebra J. J. Rabbit Peyer's patches, appendix, and popliteal lymph node B lymphocytes: a comparative analysis of their membrane immunoglobulin components and plasma cell precursor potential. J Immunol. 1975 Jan;114(1 Pt 2):492–502. [PubMed] [Google Scholar]
  6. Faulk W. P., McCormick J. N., Goodman J. R., Yoffey J. M., Fudenberg H. H. Peyer's patches: morphologic studies. Cell Immunol. 1970 Nov;1(5):500–520. doi: 10.1016/0008-8749(70)90038-9. [DOI] [PubMed] [Google Scholar]
  7. Finkelstein R. A., LoSpalluto J. J. Pathogenesis of experimental cholera. Preparation and isolation of choleragen and choleragenoid. J Exp Med. 1969 Jul 1;130(1):185–202. doi: 10.1084/jem.130.1.185. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Forsgren A., Sjöquist J. "Protein A" from S. aureus. I. Pseudo-immune reaction with human gamma-globulin. J Immunol. 1966 Dec;97(6):822–827. [PubMed] [Google Scholar]
  9. Fubara E. S., Freter R. Source and protective function of coproantibodies in intestinal disease. Am J Clin Nutr. 1972 Dec;25(12):1357–1363. doi: 10.1093/ajcn/25.12.1357. [DOI] [PubMed] [Google Scholar]
  10. Guy-Grand D., Griscelli C., Vassalli P. The gut-associated lymphoid system: nature and properties of the large dividing cells. Eur J Immunol. 1974 Jun;4(6):435–443. doi: 10.1002/eji.1830040610. [DOI] [PubMed] [Google Scholar]
  11. Henry C., Faulk W. P., Kuhn L., Yoffey J. M., Fudenberg H. H. Peyer's patches: immunologic studies. J Exp Med. 1970 Jun 1;131(6):1200–1210. doi: 10.1084/jem.131.6.1200. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Holmgren J., Lönnroth I., Ouchterlony O. Immunochemical Studies of Two Cholera Toxin-Containing Standard Culture Filtrate Preparations of Vibrio cholerae. Infect Immun. 1971 Jun;3(6):747–755. doi: 10.1128/iai.3.6.747-755.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Holmgren J., Svennerholm A. M. Enzyme-linked immunosorbent assays for cholera serology. Infect Immun. 1973 May;7(5):759–763. doi: 10.1128/iai.7.5.759-763.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Holmgren J., Svennerholm A. M., Ouchterlony O., Anderson A., Walletström G., Westerberg-Berndtsson U. Antitoxic immunity in experimental cholera: protection, and serum and local antibody responses in rabbits after enteral and parenteral immunization. Infect Immun. 1975 Dec;12(6):1331–1340. doi: 10.1128/iai.12.6.1331-1340.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Holmgren J., Svennerholm A. M., Ouchterlony O. Experimental studies on cholera immunization. 1. The response of neutralizing and vibriocidal antibodies in rabbits after immunization with culture filtrate material from V. cholerae. Acta Pathol Microbiol Scand B Microbiol Immunol. 1972;80(4):489–496. [PubMed] [Google Scholar]
  16. Lehmann J. D., Smith J. W., Miller T. E., Barnett J. A., Sanford J. P. Local immune response in experimental pyelonephritis. J Clin Invest. 1969 Nov;47(11):2541–2550. doi: 10.1172/JCI105936. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Lind I., Mansa B. Further investigation of specific and non-specific adsorption of serum globulins to Staphylococcus aureus. Acta Pathol Microbiol Scand. 1968;73(4):637–645. doi: 10.1111/j.1699-0463.1968.tb03221.x. [DOI] [PubMed] [Google Scholar]
  18. Mancini G., Carbonara A. O., Heremans J. F. Immunochemical quantitation of antigens by single radial immunodiffusion. Immunochemistry. 1965 Sep;2(3):235–254. doi: 10.1016/0019-2791(65)90004-2. [DOI] [PubMed] [Google Scholar]
  19. Pierce N. F., Gowans J. L. Cellular kinetics of the intestinal immune response to cholera toxoid in rats. J Exp Med. 1975 Dec 1;142(6):1550–1563. doi: 10.1084/jem.142.6.1550. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smiley J. D., Sachs C., Ziff M. In vitro synthesis of immunoglobulin by rheumatoid synovial membrane. J Clin Invest. 1968 Mar;47(3):624–632. doi: 10.1172/JCI105758. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Svennerholm A. M. Experimental studies on cholera immunization. 4. The antibody response to formalinized Vibrio cholerae and purified endotoxin with special reference to protective capacity. Int Arch Allergy Appl Immunol. 1975;49(4):434–452. [PubMed] [Google Scholar]
  22. Svennerholm A. M., Holmgren J. Synergistic protective effect in rabbits of immunization with Vibrio cholerae lipopolysaccharide and toxin/toxoid. Infect Immun. 1976 Mar;13(3):735–740. doi: 10.1128/iai.13.3.735-740.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Tomasi T. B., Jr, Bienenstock J. Secretory immunoglobulins. Adv Immunol. 1968;9:1–96. doi: 10.1016/s0065-2776(08)60441-1. [DOI] [PubMed] [Google Scholar]
  24. Veldkamp J., van der Gaag R., Willers J. M. The role of Peyer's patch cells in antibody formation. Immunology. 1973 Nov;25(5):761–771. [PMC free article] [PubMed] [Google Scholar]
  25. Vitetta E. S., McWilliams M., Phillips-Quagliata J. M., Lamm M. E., Uhr J. W. Cell surface immunoglobulin. XIV. Synthesis, surface expression, and secretion of immunoglobulin by Peyer's patch cells in the mouse. J Immunol. 1975 Aug;115(2):603–605. [PubMed] [Google Scholar]
  26. WADSWORTH C. A slide microtechnique for the analysis of immune precipitates in gel. Int Arch Allergy Appl Immunol. 1957;10(6):355–360. doi: 10.1159/000228394. [DOI] [PubMed] [Google Scholar]
  27. Waksman B. H., Ozer H., Blythman H. E. Appendix and M-antibody formation. VI. The functional anatomy of the rabbit appendix. Lab Invest. 1973 May;28(5):614–626. [PubMed] [Google Scholar]

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