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. 1996 Apr;64(4):1272–1283. doi: 10.1128/iai.64.4.1272-1283.1996

Salivary, nasal, genital, and systemic antibody responses in monkeys immunized intranasally with a bacterial protein antigen and the Cholera toxin B subunit.

M W Russell 1, Z Moldoveanu 1, P L White 1, G J Sibert 1, J Mestecky 1, M Michalek S 1
PMCID: PMC173915  PMID: 8606090

Abstract

Previous attempts to induce mucosal antibodies in rhesus monkeys by enteric immunization have resulted in only modest and short-lived responses, dominated by immunoglobulin M (IgM) antibodies in the plasma. In this study, two groups of rhesus monkeys were immunized intranasally three times at 2-week intervals with a bacterial protein antigen (AgI/II) either chemically coupled to or mixed with the B subunit of cholera toxin (CT), a known potent mucosal immunogen and carrier for other immunogens. Cells secreting antibodies, predominantly of the IgA isotype, to AgI/II and to CT were detected in the peripheral blood 1 week after each immunization, indicating the dissemination of IgA-secreting precursor cells through the mucosal immune system. IgG and, to a lesser extent, IgA antibodies to both proteins were induced in the plasma commencing after the second immunization. Plasma IgE concentrations and IgE antibody levels were not consistently raised during the immunization period. IgA antibodies were found in nasal and vaginal washes. Nasal IgG but not IgA antibodies showed a significant positive correlation with plasma IgG antibody levels, suggesting that they were largely derived by transudation from the circulation. Analysis of the molecular form of vaginal IgA indicated that both secretory and monomeric forms of IgA were present in various proportions. Furthermore, neither IgG nor IgA antibodies in vaginal washes were correlated with plasma antibody responses, suggesting the contribution of locally synthesized antibodies of both isotypes. Comparison of the responses between the two groups of animals showed only sporadic significant differences, indicating that intranasal immunization with AgI/II either coupled to or mixed with the B subunit of CT was equally effective at inducing generalized IgA antibody responses in the mucosal immune system and predominantly IgG antibodies in the plasma.

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Selected References

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  1. Bessen D., Fischetti V. A. Influence of intranasal immunization with synthetic peptides corresponding to conserved epitopes of M protein on mucosal colonization by group A streptococci. Infect Immun. 1988 Oct;56(10):2666–2672. doi: 10.1128/iai.56.10.2666-2672.1988. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Brandtzaeg P., Halstensen T. S. Immunology and immunopathology of tonsils. Adv Otorhinolaryngol. 1992;47:64–75. doi: 10.1159/000421721. [DOI] [PubMed] [Google Scholar]
  3. Brandtzaeg P. Immunocompetent cells of the upper airway: functions in normal and diseased mucosa. Eur Arch Otorhinolaryngol. 1995;252 (Suppl 1):S8–21. doi: 10.1007/BF02484429. [DOI] [PubMed] [Google Scholar]
  4. Bélec L., Tévi-Bénissan C., Lu X. S., Prazuck T., Pillot J. Local synthesis of IgG antibodies to HIV within the female and male genital tracts during asymptomatic and pre-AIDS stages of HIV infection. AIDS Res Hum Retroviruses. 1995 Jun;11(6):719–729. doi: 10.1089/aid.1995.11.719. [DOI] [PubMed] [Google Scholar]
  5. Challacombe S. J., Russell M. W. Estimation of the intravascular half-lives of normal rhesus monkey IgG, IgA and IgM. Immunology. 1979 Feb;36(2):331–338. [PMC free article] [PubMed] [Google Scholar]
  6. Czerkinsky C., Moldoveanu Z., Mestecky J., Nilsson L. A., Ouchterlony O. A novel two colour ELISPOT assay. I. Simultaneous detection of distinct types of antibody-secreting cells. J Immunol Methods. 1988 Nov 25;115(1):31–37. doi: 10.1016/0022-1759(88)90306-7. [DOI] [PubMed] [Google Scholar]
  7. Czerkinsky C., Prince S. J., Michalek S. M., Jackson S., Russell M. W., Moldoveanu Z., McGhee J. R., Mestecky J. IgA antibody-producing cells in peripheral blood after antigen ingestion: evidence for a common mucosal immune system in humans. Proc Natl Acad Sci U S A. 1987 Apr;84(8):2449–2453. doi: 10.1073/pnas.84.8.2449. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Czerkinsky C., Russell M. W., Lycke N., Lindblad M., Holmgren J. Oral administration of a streptococcal antigen coupled to cholera toxin B subunit evokes strong antibody responses in salivary glands and extramucosal tissues. Infect Immun. 1989 Apr;57(4):1072–1077. doi: 10.1128/iai.57.4.1072-1077.1989. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Czerkinsky C., Svennerholm A. M., Quiding M., Jonsson R., Holmgren J. Antibody-producing cells in peripheral blood and salivary glands after oral cholera vaccination of humans. Infect Immun. 1991 Mar;59(3):996–1001. doi: 10.1128/iai.59.3.996-1001.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Dertzbaugh M. T., Peterson D. L., Macrina F. L. Cholera toxin B-subunit gene fusion: structural and functional analysis of the chimeric protein. Infect Immun. 1990 Jan;58(1):70–79. doi: 10.1128/iai.58.1.70-79.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Elson C. O. Cholera toxin and its subunits as potential oral adjuvants. Curr Top Microbiol Immunol. 1989;146:29–33. doi: 10.1007/978-3-642-74529-4_3. [DOI] [PubMed] [Google Scholar]
  12. Ferguson A., Humphreys K. A., Croft N. M. Technical report: results of immunological tests on faecal extracts are likely to be extremely misleading. Clin Exp Immunol. 1995 Jan;99(1):70–75. doi: 10.1111/j.1365-2249.1995.tb03474.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Hajishengallis G., Hollingshead S. K., Koga T., Russell M. W. Mucosal immunization with a bacterial protein antigen genetically coupled to cholera toxin A2/B subunits. J Immunol. 1995 May 1;154(9):4322–4332. [PubMed] [Google Scholar]
  14. Hameleers D. M., van der Ven I., Biewenga J., Sminia T. Mucosal and systemic antibody formation in the rat after intranasal administration of three different antigens. Immunol Cell Biol. 1991 Apr;69(Pt 2):119–125. doi: 10.1038/icb.1991.18. [DOI] [PubMed] [Google Scholar]
  15. Harkema J. R., Plopper C. G., Hyde D. M., Wilson D. W., St George J. A., Wong V. J. Nonolfactory surface epithelium of the nasal cavity of the bonnet monkey: a morphologic and morphometric study of the transitional and respiratory epithelium. Am J Anat. 1987 Nov;180(3):266–279. doi: 10.1002/aja.1001800308. [DOI] [PubMed] [Google Scholar]
  16. Hirabayashi Y., Kurata H., Funato H., Nagamine T., Aizawa C., Tamura S., Shimada K., Kurata T. Comparison of intranasal inoculation of influenza HA vaccine combined with cholera toxin B subunit with oral or parenteral vaccination. Vaccine. 1990 Jun;8(3):243–248. doi: 10.1016/0264-410x(90)90053-o. [DOI] [PubMed] [Google Scholar]
  17. Holmgren J., Lycke N., Czerkinsky C. Cholera toxin and cholera B subunit as oral-mucosal adjuvant and antigen vector systems. Vaccine. 1993 Sep;11(12):1179–1184. doi: 10.1016/0264-410x(93)90039-z. [DOI] [PubMed] [Google Scholar]
  18. Jagusztyn-Krynicka E. K., Clark-Curtiss J. E., Curtiss R., 3rd Escherichia coli heat-labile toxin subunit B fusions with Streptococcus sobrinus antigens expressed by Salmonella typhimurium oral vaccine strains: importance of the linker for antigenicity and biological activities of the hybrid proteins. Infect Immun. 1993 Mar;61(3):1004–1015. doi: 10.1128/iai.61.3.1004-1015.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Jertborn M., Svennerholm A. M., Holmgren J. Safety and immunogenicity of an oral recombinant cholera B subunit-whole cell vaccine in Swedish volunteers. Vaccine. 1992;10(2):130–132. doi: 10.1016/0264-410x(92)90030-n. [DOI] [PubMed] [Google Scholar]
  20. Kantele A., Arvilommi H., Jokinen I. Specific immunoglobulin-secreting human blood cells after peroral vaccination against Salmonella typhi. J Infect Dis. 1986 Jun;153(6):1126–1131. doi: 10.1093/infdis/153.6.1126. [DOI] [PubMed] [Google Scholar]
  21. Katz J., Harmon C. C., Buckner G. P., Richardson G. J., Russell M. W., Michalek S. M. Protective salivary immunoglobulin A responses against Streptococcus mutans infection after intranasal immunization with S. mutans antigen I/II coupled to the B subunit of cholera toxin. Infect Immun. 1993 May;61(5):1964–1971. doi: 10.1128/iai.61.5.1964-1971.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Kuper C. F., Koornstra P. J., Hameleers D. M., Biewenga J., Spit B. J., Duijvestijn A. M., van Breda Vriesman P. J., Sminia T. The role of nasopharyngeal lymphoid tissue. Immunol Today. 1992 Jun;13(6):219–224. doi: 10.1016/0167-5699(92)90158-4. [DOI] [PubMed] [Google Scholar]
  23. Lehner T., Bergmeier L. A., Panagiotidi C., Tao L., Brookes R., Klavinskis L. S., Walker P., Walker J., Ward R. G., Hussain L. Induction of mucosal and systemic immunity to a recombinant simian immunodeficiency viral protein. Science. 1992 Nov 20;258(5086):1365–1369. doi: 10.1126/science.1360702. [DOI] [PubMed] [Google Scholar]
  24. Lehner T., Bergmeier L. A., Tao L., Panagiotidi C., Klavinskis L. S., Hussain L., Ward R. G., Meyers N., Adams S. E., Gearing A. J. Targeted lymph node immunization with simian immunodeficiency virus p27 antigen to elicit genital, rectal, and urinary immune responses in nonhuman primates. J Immunol. 1994 Aug 15;153(4):1858–1868. [PubMed] [Google Scholar]
  25. Lewis D. J., Novotny P., Dougan G., Griffin G. E. The early cellular and humoral immune response to primary and booster oral immunization with cholera toxin B subunit. Eur J Immunol. 1991 Sep;21(9):2087–2094. doi: 10.1002/eji.1830210917. [DOI] [PubMed] [Google Scholar]
  26. 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]
  27. Marinaro M., Staats H. F., Hiroi T., Jackson R. J., Coste M., Boyaka P. N., Okahashi N., Yamamoto M., Kiyono H., Bluethmann H. Mucosal adjuvant effect of cholera toxin in mice results from induction of T helper 2 (Th2) cells and IL-4. J Immunol. 1995 Nov 15;155(10):4621–4629. [PubMed] [Google Scholar]
  28. Marx P. A., Compans R. W., Gettie A., Staas J. K., Gilley R. M., Mulligan M. J., Yamshchikov G. V., Chen D., Eldridge J. H. Protection against vaginal SIV transmission with microencapsulated vaccine. Science. 1993 May 28;260(5112):1323–1327. doi: 10.1126/science.8493576. [DOI] [PubMed] [Google Scholar]
  29. McKenzie S. J., Halsey J. F. Cholera toxin B subunit as a carrier protein to stimulate a mucosal immune response. J Immunol. 1984 Oct;133(4):1818–1824. [PubMed] [Google Scholar]
  30. Menge A. C., Michalek S. M., Russell M. W., Mestecky J. Immune response of the female rat genital tract after oral and local immunization with keyhole limpet hemocyanin conjugated to the cholera toxin B subunit. Infect Immun. 1993 May;61(5):2162–2171. doi: 10.1128/iai.61.5.2162-2171.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Mestecky J., Lue C., Russell M. W. Selective transport of IgA. Cellular and molecular aspects. Gastroenterol Clin North Am. 1991 Sep;20(3):441–471. [PubMed] [Google Scholar]
  32. Mestecky J. The common mucosal immune system and current strategies for induction of immune responses in external secretions. J Clin Immunol. 1987 Jul;7(4):265–276. doi: 10.1007/BF00915547. [DOI] [PubMed] [Google Scholar]
  33. Michalek S. M., Lackner A. A., Katz J., Russell M. W., Eldridge J. H., Mestecky J., Lallone R., Moldoveanu Z. Oral immunization studies with Streptococcus mutans and influenza vaccines in rhesus macaque monkeys. Adv Exp Med Biol. 1995;371B:1423–1429. [PubMed] [Google Scholar]
  34. Miller C. J., Kang D. W., Marthas M., Moldoveanu Z., Kiyono H., Marx P., Eldridge J. H., Mestecky J., McGhee J. R. Genital secretory immune response to chronic simian immunodeficiency virus (SIV) infection: a comparison between intravenously and genitally inoculated rhesus macaques. Clin Exp Immunol. 1992 Jun;88(3):520–526. doi: 10.1111/j.1365-2249.1992.tb06481.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Moldoveanu Z., Clements M. L., Prince S. J., Murphy B. R., Mestecky J. Human immune responses to influenza virus vaccines administered by systemic or mucosal routes. Vaccine. 1995 Aug;13(11):1006–1012. doi: 10.1016/0264-410x(95)00016-t. [DOI] [PubMed] [Google Scholar]
  36. Moldoveanu Z., Russell M. W., Wu H. Y., Huang W. Q., Compans R. W., Mestecky J. Compartmentalization within the common mucosal immune system. Adv Exp Med Biol. 1995;371A:97–101. doi: 10.1007/978-1-4615-1941-6_17. [DOI] [PubMed] [Google Scholar]
  37. Nair P. N., Rossinsky K. Organization of lymphoid tissue in the tonsilla lingualis. An ultrastructural study in Macaca fascicularis (Primates, Cercopithecoidea). Cell Tissue Res. 1985;240(1):233–242. doi: 10.1007/BF00217579. [DOI] [PubMed] [Google Scholar]
  38. Nedrud J. G., Liang X. P., Hague N., Lamm M. E. Combined oral/nasal immunization protects mice from Sendai virus infection. J Immunol. 1987 Nov 15;139(10):3484–3492. [PubMed] [Google Scholar]
  39. Ogra P. L. Effect of tonsillectomy and adenoidectomy on nasopharyngeal antibody response to poliovirus. N Engl J Med. 1971 Jan 14;284(2):59–64. doi: 10.1056/NEJM197101142840201. [DOI] [PubMed] [Google Scholar]
  40. Ogra P. L., Ogra S. S. Local antibody response to poliovaccine in the human female genital tract. J Immunol. 1973 May;110(5):1307–1311. [PubMed] [Google Scholar]
  41. Quiding-Järbrink M., Granström G., Nordström I., Holmgren J., Czerkinsky C. Induction of compartmentalized B-cell responses in human tonsils. Infect Immun. 1995 Mar;63(3):853–857. doi: 10.1128/iai.63.3.853-857.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  42. Quiding-Järbrink M., Lakew M., Nordström I., Banchereau J., Butcher E., Holmgren J., Czerkinsky C. Human circulating specific antibody-forming cells after systemic and mucosal immunizations: differential homing commitments and cell surface differentiation markers. Eur J Immunol. 1995 Feb;25(2):322–327. doi: 10.1002/eji.1830250203. [DOI] [PubMed] [Google Scholar]
  43. Richardson J., Kaushic C., Wira C. R. Estradiol regulation of secretory component: expression by rat uterine epithelial cells. J Steroid Biochem Mol Biol. 1993 Dec;47(1-6):143–149. doi: 10.1016/0960-0760(93)90068-8. [DOI] [PubMed] [Google Scholar]
  44. Russell M. W., Bergmeier L. A., Zanders E. D., Lehner T. Protein antigens of Streptococcus mutans: purification and properties of a double antigen and its protease-resistant component. Infect Immun. 1980 May;28(2):486–493. doi: 10.1128/iai.28.2.486-493.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Russell M. W., Brown T. A., Radl J., Haaijman J. J., Mestecky J. Assay of human IgA subclass antibodies in serum and secretions by means of monoclonal antibodies. J Immunol Methods. 1986 Feb 27;87(1):87–93. doi: 10.1016/0022-1759(86)90347-9. [DOI] [PubMed] [Google Scholar]
  46. Russell M. W., Wu H. Y. Distribution, persistence, and recall of serum and salivary antibody responses to peroral immunization with protein antigen I/II of Streptococcus mutans coupled to the cholera toxin B subunit. Infect Immun. 1991 Nov;59(11):4061–4070. doi: 10.1128/iai.59.11.4061-4070.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Snider D. P., Marshall J. S., Perdue M. H., Liang H. Production of IgE antibody and allergic sensitization of intestinal and peripheral tissues after oral immunization with protein Ag and cholera toxin. J Immunol. 1994 Jul 15;153(2):647–657. [PubMed] [Google Scholar]
  48. Sun J. B., Holmgren J., Czerkinsky C. Cholera toxin B subunit: an efficient transmucosal carrier-delivery system for induction of peripheral immunological tolerance. Proc Natl Acad Sci U S A. 1994 Nov 8;91(23):10795–10799. doi: 10.1073/pnas.91.23.10795. [DOI] [PMC free article] [PubMed] [Google Scholar]
  49. Takahashi I., Okahashi N., Kanamoto T., Asakawa H., Koga T. Intranasal immunization of mice with recombinant protein antigen of serotype c Streptococcus mutans and cholera toxin B subunit. Arch Oral Biol. 1990;35(6):475–477. doi: 10.1016/0003-9969(90)90211-r. [DOI] [PubMed] [Google Scholar]
  50. Tamura S., Asanuma H., Tomita T., Komase K., Kawahara K., Danbara H., Hattori N., Watanabe K., Suzuki Y., Nagamine T. Escherichia coli heat-labile enterotoxin B subunits supplemented with a trace amount of the holotoxin as an adjuvant for nasal influenza vaccine. Vaccine. 1994 Sep;12(12):1083–1089. doi: 10.1016/0264-410x(94)90177-5. [DOI] [PubMed] [Google Scholar]
  51. Tamura S., Shoji Y., Hasiguchi K., Aizawa C., Kurata T. Effects of cholera toxin adjuvant on IgE antibody response to orally or nasally administered ovalbumin. Vaccine. 1994 Oct;12(13):1238–1240. doi: 10.1016/0264-410x(94)90250-x. [DOI] [PubMed] [Google Scholar]
  52. Taylor L. D., Daniels C. K., Schmucker D. L. Ageing compromises gastrointestinal mucosal immune response in the rhesus monkey. Immunology. 1992 Apr;75(4):614–618. [PMC free article] [PubMed] [Google Scholar]
  53. Thapar M. A., Parr E. L., Parr M. B. Secretory immune responses in mouse vaginal fluid after pelvic, parenteral or vaginal immunization. Immunology. 1990 May;70(1):121–125. [PMC free article] [PubMed] [Google Scholar]
  54. Wagner D. K., Clements M. L., Reimer C. B., Snyder M., Nelson D. L., Murphy B. R. Analysis of immunoglobulin G antibody responses after administration of live and inactivated influenza A vaccine indicates that nasal wash immunoglobulin G is a transudate from serum. J Clin Microbiol. 1987 Mar;25(3):559–562. doi: 10.1128/jcm.25.3.559-562.1987. [DOI] [PMC free article] [PubMed] [Google Scholar]
  55. Walker R. I. New strategies for using mucosal vaccination to achieve more effective immunization. Vaccine. 1994 Apr;12(5):387–400. doi: 10.1016/0264-410x(94)90112-0. [DOI] [PubMed] [Google Scholar]
  56. Wira C. R., Sandoe C. P. Effect of uterine immunization and oestradiol on specific IgA and IgG antibodies in uterine, vaginal and salivary secretions. Immunology. 1989 Sep;68(1):24–30. [PMC free article] [PubMed] [Google Scholar]
  57. Wu H. Y., Russell M. W. Induction of mucosal immunity by intranasal application of a streptococcal surface protein antigen with the cholera toxin B subunit. Infect Immun. 1993 Jan;61(1):314–322. doi: 10.1128/iai.61.1.314-322.1993. [DOI] [PMC free article] [PubMed] [Google Scholar]
  58. van der Ven I., Sminia T. The development and structure of mouse nasal-associated lymphoid tissue: an immuno- and enzyme-histochemical study. Reg Immunol. 1993 Mar-Apr;5(2):69–75. [PubMed] [Google Scholar]

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