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. 1991 Oct;59(10):3630–3638. doi: 10.1128/iai.59.10.3630-3638.1991

Host defense against cholera toxin is strongly CD4+ T cell dependent.

E Hörnqvist 1, T J Goldschmidt 1, R Holmdahl 1, N Lycke 1
PMCID: PMC258931  PMID: 1910010

Abstract

This study investigates the role of CD4+ T cells in host defense against cholera enterotoxin-induced diarrhea. Antitoxin immunoglobulin A formation and gut protection against cholera toxin (CT) following oral immunizations with CT were evaluated in normal mice and mice that had been depleted of CD4+ T cells by in vivo treatment with specific anti-CD4 monoclonal antibodies. Flow cytometer analysis demonstrated that anti-CD4 monoclonal antibody effectively eliminated CD4+ T cells in the spleen, mesenteric lymph nodes, and Peyer's patches. In contrast, lamina propria lymphocytes demonstrated only some decrease in CD4+ T-cell numbers following antibody treatment. However, CD4 expression of individual lamina propria lymphocytes was strongly down-regulated. Depletion of CD4+ T cells performed prior to oral immunization with CT completely inhibited the ability to respond to CT. No antitoxin production, as detected at the single-cell level by the ELISPOT technique, was found in the spleen, mesenteric lymph nodes, or Peyer's patches, nor did we observe serum antitoxin responses in these mice. Control mice demonstrated strong antitoxin responses in all locations following oral immunization with CT. Anti-CD4 antibody treatment also effectively inhibited the antitoxin immunoglobulin A response in the lamina propria to CT as well as blocked the ability to develop gut protection against CT challenge of ligated intestinal loops after oral CT immunization. Thus, in vivo CD4+ T-cell depletion rendered these mice unable to develop protective immunity in the gut following oral immunization with CT. Moreover, CD4+ T-cell depletion effectively inhibited the antitoxin immune response in the gut lamina propria, mesenteric lymph nodes, Peyer's patches, and spleen when performed prior to both priming and booster immunizations with CT. This study clearly demonstrates the requirement of functional CD4+ T cells in the gut immune system for the development of host defense against CT-induced disease. Our data also reinforce the concept of a strong association between gut protection against CT and local production of neutralizing immunoglobulin A antitoxin.

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Selected References

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  1. Beagley K. W., Eldridge J. H., Lee F., Kiyono H., Everson M. P., Koopman W. J., Hirano T., Kishimoto T., McGhee J. R. Interleukins and IgA synthesis. Human and murine interleukin 6 induce high rate IgA secretion in IgA-committed B cells. J Exp Med. 1989 Jun 1;169(6):2133–2148. doi: 10.1084/jem.169.6.2133. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bergstedt-Lindqvist S., Moon H. B., Persson U., Möller G., Heusser C., Severinson E. Interleukin 4 instructs uncommitted B lymphocytes to switch to IgG1 and IgE. Eur J Immunol. 1988 Jul;18(7):1073–1077. doi: 10.1002/eji.1830180716. [DOI] [PubMed] [Google Scholar]
  3. Blalock J. E. A molecular basis for bidirectional communication between the immune and neuroendocrine systems. Physiol Rev. 1989 Jan;69(1):1–32. doi: 10.1152/physrev.1989.69.1.1. [DOI] [PubMed] [Google Scholar]
  4. Brandtzaeg P., Bosnes V., Halstensen T. S., Scott H., Sollid L. M., Valnes K. N. T lymphocytes in human gut epithelium preferentially express the alpha/beta antigen receptor and are often CD45/UCHL1-positive. Scand J Immunol. 1989 Jul;30(1):123–128. doi: 10.1111/j.1365-3083.1989.tb01196.x. [DOI] [PubMed] [Google Scholar]
  5. Brandtzaeg P., Halstensen T. S., Kett K., Krajci P., Kvale D., Rognum T. O., Scott H., Sollid L. M. Immunobiology and immunopathology of human gut mucosa: humoral immunity and intraepithelial lymphocytes. Gastroenterology. 1989 Dec;97(6):1562–1584. doi: 10.1016/0016-5085(89)90406-x. [DOI] [PubMed] [Google Scholar]
  6. Charlton B., Burkhardt K., Mandel T. E. How important is the L3T4 antigen to L3T4+ cell function in vivo? Immunol Today. 1988 Jun;9(6):165–167. doi: 10.1016/0167-5699(88)91290-X. [DOI] [PubMed] [Google Scholar]
  7. Cobbold S. P., Jayasuriya A., Nash A., Prospero T. D., Waldmann H. Therapy with monoclonal antibodies by elimination of T-cell subsets in vivo. Nature. 1984 Dec 6;312(5994):548–551. doi: 10.1038/312548a0. [DOI] [PubMed] [Google Scholar]
  8. Coulie P. G., Coutelier J. P., Uyttenhove C., Lambotte P., Van Snick J. In vivo suppression of T-dependent antibody responses by treatment with a monoclonal anti-L3T4 antibody. Eur J Immunol. 1985 Jun;15(6):638–640. doi: 10.1002/eji.1830150620. [DOI] [PubMed] [Google Scholar]
  9. Czerkinsky C. C., Nilsson L. A., Nygren H., Ouchterlony O., Tarkowski A. A solid-phase enzyme-linked immunospot (ELISPOT) assay for enumeration of specific antibody-secreting cells. J Immunol Methods. 1983 Dec 16;65(1-2):109–121. doi: 10.1016/0022-1759(83)90308-3. [DOI] [PubMed] [Google Scholar]
  10. Goldschmidt T. J., Holmdahl R., Klareskog L. Depletion of murine T cells by in vivo monoclonal antibody treatment is enhanced by adding an autologous anti-rat kappa chain antibody. J Immunol Methods. 1988 Jul 22;111(2):219–226. doi: 10.1016/0022-1759(88)90130-5. [DOI] [PubMed] [Google Scholar]
  11. Harriman G. R., Kunimoto D. Y., Elliott J. F., Paetkau V., Strober W. The role of IL-5 in IgA B cell differentiation. J Immunol. 1988 May 1;140(9):3033–3039. [PubMed] [Google Scholar]
  12. Holmgren J., Fryklund J., Larsson H. Gamma-interferon-mediated down-regulation of electrolyte secretion by intestinal epithelial cells: a local immune mechanism? Scand J Immunol. 1989 Oct;30(4):499–503. doi: 10.1111/j.1365-3083.1989.tb02456.x. [DOI] [PubMed] [Google Scholar]
  13. Howard M., Farrar J., Hilfiker M., Johnson B., Takatsu K., Hamaoka T., Paul W. E. Identification of a T cell-derived b cell growth factor distinct from interleukin 2. J Exp Med. 1982 Mar 1;155(3):914–923. doi: 10.1084/jem.155.3.914. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Husband A. J. Kinetics of extravasation and redistribution of IgA-specific antibody-containing cells in the intestine. J Immunol. 1982 Mar;128(3):1355–1359. [PubMed] [Google Scholar]
  15. James S. P., Graeff A. S. Effect of IL-2 on immunoregulatory function of intestinal lamina propria T cells in normal non-human primates. Clin Exp Immunol. 1987 Nov;70(2):394–402. [PMC free article] [PubMed] [Google Scholar]
  16. Janossy G., Bofill M., Rowe D., Muir J., Beverley P. C. The tissue distribution of T lymphocytes expressing different CD45 polypeptides. Immunology. 1989 Apr;66(4):517–525. [PMC free article] [PubMed] [Google Scholar]
  17. Lange S., Holmgren J. Protective antitoxic cholera immunity in mice: influence of route and number of immunizations and mode of action of protective antibodies. Acta Pathol Microbiol Scand C. 1978 Aug;86C(4):145–152. doi: 10.1111/j.1699-0463.1978.tb02572.x. [DOI] [PubMed] [Google Scholar]
  18. Lange S., Lönnroth I., Nygren H. Protection against experimental cholera in the rat. A study on the formation of antibodies against cholera toxin and desensitization of adenylate cyclase after immunization with cholera toxin. Int Arch Allergy Appl Immunol. 1984;75(2):143–148. [PubMed] [Google Scholar]
  19. Lanier L. L., Gutman G. A., Lewis D. E., Griswold S. T., Warner N. L. Monoclonal antibodies against rat immunoglobulin kappa chains. Hybridoma. 1982;1(2):125–131. doi: 10.1089/hyb.1.1982.1.125. [DOI] [PubMed] [Google Scholar]
  20. Lebman D. A., Nomura D. Y., Coffman R. L., Lee F. D. Molecular characterization of germ-line immunoglobulin A transcripts produced during transforming growth factor type beta-induced isotype switching. Proc Natl Acad Sci U S A. 1990 May;87(10):3962–3966. doi: 10.1073/pnas.87.10.3962. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Lycke N. A sensitive method for the detection of specific antibody production in different isotypes from single lamina propria plasma cells. Scand J Immunol. 1986 Oct;24(4):393–403. doi: 10.1111/j.1365-3083.1986.tb02127.x. [DOI] [PubMed] [Google Scholar]
  22. Lycke N., Eriksen L., Holmgren J. Protection against cholera toxin after oral immunization is thymus-dependent and associated with intestinal production of neutralizing IgA antitoxin. Scand J Immunol. 1987 Apr;25(4):413–419. doi: 10.1111/j.1365-3083.1987.tb02208.x. [DOI] [PubMed] [Google Scholar]
  23. Lycke N., Holmgren J. Long-term cholera antitoxin memory in the gut can be triggered to antibody formation associated with protection within hours of an oral challenge immunization. Scand J Immunol. 1987 Apr;25(4):407–412. doi: 10.1111/j.1365-3083.1987.tb02207.x. [DOI] [PubMed] [Google Scholar]
  24. Lycke N., Lindholm L., Holmgren J. Cholera antibody production in vitro by peripheral blood lymphocytes following oral immunization of humans and mice. Clin Exp Immunol. 1985 Oct;62(1):39–47. [PMC free article] [PubMed] [Google Scholar]
  25. Lycke N., Lindholm L., Holmgren J. IgA isotype restriction in the mucosal but not in the extramucosal immune response after oral immunizations with cholera toxin or cholera B subunit. Int Arch Allergy Appl Immunol. 1983;72(2):119–127. doi: 10.1159/000234853. [DOI] [PubMed] [Google Scholar]
  26. Lönnroth I., Lange S., Skadhauge E. The antisecretory factors: inducible proteins which modulate secretion in the small intestine. Comp Biochem Physiol A Comp Physiol. 1988;90(4):611–617. doi: 10.1016/0300-9629(88)90675-5. [DOI] [PubMed] [Google Scholar]
  27. Pierce N. F., Cray W. C., Jr Determinants of the localization, magnitude, and duration of a specific mucosal IgA plasma cell response in enterically immunized rats. J Immunol. 1982 Mar;128(3):1311–1315. [PubMed] [Google Scholar]
  28. Pierres A., Naquet P., Van Agthoven A., Bekkhoucha F., Denizot F., Mishal Z., Schmitt-Verhulst A. M., Pierres M. A rat anti-mouse T4 monoclonal antibody (H129.19) inhibits the proliferation of Ia-reactive T cell clones and delineates two phenotypically distinct (T4+, Lyt-2,3-, and T4-, Lyt-2,3+) subsets among anti-Ia cytolytic T cell clones. J Immunol. 1984 Jun;132(6):2775–2782. [PubMed] [Google Scholar]
  29. Quiding M., Nordström I., Kilander A., Andersson G., Hanson L. A., Holmgren J., Czerkinsky C. Intestinal immune responses in humans. Oral cholera vaccination induces strong intestinal antibody responses and interferon-gamma production and evokes local immunological memory. J Clin Invest. 1991 Jul;88(1):143–148. doi: 10.1172/JCI115270. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. Ranges G. E., Cooper S. M., Sriram S. In vivo immunomodulation by monoclonal anti-L3T4. 1. Effects on humoral and cell-mediated immune response. Cell Immunol. 1987 Apr 15;106(1):163–173. doi: 10.1016/0008-8749(87)90159-6. [DOI] [PubMed] [Google Scholar]
  31. Snapper C. M., Finkelman F. D., Paul W. E. Differential regulation of IgG1 and IgE synthesis by interleukin 4. J Exp Med. 1988 Jan 1;167(1):183–196. doi: 10.1084/jem.167.1.183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Svennerholm A. M., Jertborn M., Gothefors L., Karim A. M., Sack D. A., Holmgren J. Mucosal antitoxic and antibacterial immunity after cholera disease and after immunization with a combined B subunit-whole cell vaccine. J Infect Dis. 1984 Jun;149(6):884–893. doi: 10.1093/infdis/149.6.884. [DOI] [PubMed] [Google Scholar]
  33. Tam M. R., Bernstein I. D., Nowinski R. C. Alteration of lymphoid cells in AKR mice by treatment with monoclonal antibody against Thy-1 antigen. Transplantation. 1982 Mar;33(3):269–273. doi: 10.1097/00007890-198203000-00012. [DOI] [PubMed] [Google Scholar]
  34. Wofsy D., Mayes D. C., Woodcock J., Seaman W. E. Inhibition of humoral immunity in vivo by monoclonal antibody to L3T4: studies with soluble antigens in intact mice. J Immunol. 1985 Sep;135(3):1698–1701. [PubMed] [Google Scholar]
  35. Wofsy D., Seaman W. E. Successful treatment of autoimmunity in NZB/NZW F1 mice with monoclonal antibody to L3T4. J Exp Med. 1985 Feb 1;161(2):378–391. doi: 10.1084/jem.161.2.378. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Zeitz M., Greene W. C., Peffer N. J., James S. P. Lymphocytes isolated from the intestinal lamina propria of normal nonhuman primates have increased expression of genes associated with T-cell activation. Gastroenterology. 1988 Mar;94(3):647–655. doi: 10.1016/0016-5085(88)90235-1. [DOI] [PubMed] [Google Scholar]

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