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. 1974 Nov 30;140(6):1604–1614. doi: 10.1084/jem.140.6.1604

IMMUNOLOGICAL RESPONSES OF MICE TO NATIVE PROTOPLASMIC POLYSACCHARIDE AND LIPOPOLYSACCHARIDE

Functional Separation of the Two Signals Required to Stimulate a Secondary Antibody Response

Kenneth B von Eschen 1, Jon A Rudbach 1
PMCID: PMC2139752  PMID: 4610079

Abstract

Functional separation of the two signals involved in stimulating immunological responses was achieved through the judicious use of two natural bacterial antigens. Native protoplasmic polysaccharide (NPP) extracted from Escherichia coli was immunochemically identical to the lipopolysaccharide (LPS) extracted from the same organism. However, NPP was not endotoxic, not mitogenic, did not fix complement, and was immunologically independent of T cells. The NPP, which appeared to contain only the antigenic signal, could induce a primary antibody response in mice and could sensitize mice for a secondary response. However, the antigenic signal contained in NPP was insufficient to trigger a secondary response in mice primed with either NPP or LPS. LPS, containing both the antigenic and second signals, was required to trigger a secondary response in primed mice.

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Selected References

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  1. ANACKER R. L., FINKELSTEIN R. A., HASKINS W. T., LANDY M., MILNER K. C., RIBI E., STASHAK P. W. ORIGIN AND PROPERTIES OF NATURALLY OCCURRING HAPTEN FROM ESCHERICHIA COLI. J Bacteriol. 1964 Dec;88:1705–1720. doi: 10.1128/jb.88.6.1705-1720.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Anacker R. L., Bickel W. D., Haskins W. T., Milner K. C., Ribi E., Rudbach J. A. Frequency of occurrence of native hapten among enterobacterial species. J Bacteriol. 1966 Apr;91(4):1427–1433. doi: 10.1128/jb.91.4.1427-1433.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Andersson B., Blomgren H. Evidence for thymus-independent humoral antibody production in mice against polyvinylpyrrolidone and E. coli lipopolysaccharide. Cell Immunol. 1971 Oct;2(5):411–424. doi: 10.1016/0008-8749(71)90052-9. [DOI] [PubMed] [Google Scholar]
  4. Andersson B. Induction of immunity and immunologic paralysis in mice against polyvinyl pyrrolidone. J Immunol. 1969 May;102(5):1309–1313. [PubMed] [Google Scholar]
  5. Andersson J., Melchers F., Galanos C., Lüderitz O. The mitogenic effect of lipopolysaccharide on bone marrow-derived mouse lymphocytes. Lipid A as the mitogenic part of the molecule. J Exp Med. 1973 Apr 1;137(4):943–953. doi: 10.1084/jem.137.4.943. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Armerding D., Katz D. H. Activation of T and B lymphocytes in vitro. II. Biological and biochemical properties of an allogeneic effect factor (AEF) active in triggering specific B lymphocytes. J Exp Med. 1974 Jul 1;140(1):19–37. doi: 10.1084/jem.140.1.19. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Baker P. J., Stashak P. W., Amsbaugh D. F., Prescott B. Characterization of the antibody response to type 3 pneumococcal polysaccharide at the cellular level. I. Dose-response studies and the effect of prior immunization on the magnitude of the antibody response. Immunology. 1971 Apr;20(4):469–480. [PMC free article] [PubMed] [Google Scholar]
  8. Bretscher P., Cohn M. A theory of self-nonself discrimination. Science. 1970 Sep 11;169(3950):1042–1049. doi: 10.1126/science.169.3950.1042. [DOI] [PubMed] [Google Scholar]
  9. Byfield P., Sercarz E. The X-Y-Z scheme of immunocyte maturation. VII. Cell division and the establishment of short-term IgM memory. J Exp Med. 1969 May 1;129(5):897–907. doi: 10.1084/jem.129.5.897. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chiller J. M., Skidmore B. J., Morrison D. C., Weigle W. O. Relationship of the structure of bacterial lipopolysaccharides to its function in mitogenesis and adjuvanticity. Proc Natl Acad Sci U S A. 1973 Jul;70(7):2129–2133. doi: 10.1073/pnas.70.7.2129. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Chiller J. M., Weigle W. O. Termination of tolerance to human gamma globulin in mice by antigen and bacterial lipopolysaccharide (endotoxin). J Exp Med. 1973 Mar 1;137(3):740–750. doi: 10.1084/jem.137.3.740. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Cohn M. The take-home lesson--1971. Ann N Y Acad Sci. 1971 Dec 31;190:529–584. doi: 10.1111/j.1749-6632.1971.tb13562.x. [DOI] [PubMed] [Google Scholar]
  13. Coutinho A., Gronowicz E., Bullock W. W., Möller G. Mechanism of thymus-independent immunocyte triggering. Mitogenic activation of B cells results in specific immune responses. J Exp Med. 1974 Jan 1;139(1):74–92. doi: 10.1084/jem.139.1.74. [DOI] [PMC free article] [PubMed] [Google Scholar]
  14. Doria G., Agarossi G., Di Pietro S. Enhancing activity of thymocyte culture cell-free medium on the in vitro immune response of spleen cells from neonatally thymectomized mice to sheep RBC. J Immunol. 1972 Jan;108(1):268–270. [PubMed] [Google Scholar]
  15. Dukor P., Hartmann K. U. Hypothesis. Bound C3 as the second signal for B-cell activation. Cell Immunol. 1973 Jun;7(3):349–356. doi: 10.1016/0008-8749(73)90199-8. [DOI] [PubMed] [Google Scholar]
  16. FUKUSHI K., ANACKER R. L., HASKINS W. T., LANDY M., MILNER K. C., RIBI E. EXTRACTION AND PURIFICATION OF ENDOTOXIN FROM ENTEROBACTERIACEAE: A COMPARISON OF SELECTED METHODS AND SOURCES. J Bacteriol. 1964 Feb;87:391–400. doi: 10.1128/jb.87.2.391-400.1964. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Feldmann M., Basten A. Cell interactions in the immune response in vitro. IV. Comparison of the effects of antigen-specific and allogeneic thymus-derived cell factors. J Exp Med. 1972 Oct 1;136(4):722–736. doi: 10.1084/jem.136.4.722. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Feldmann M., Nossal G. J. Tolerance, enhancement and the regulation of interactions between T cells, B cells and macrophages. Transplant Rev. 1972;13:3–34. doi: 10.1111/j.1600-065x.1972.tb00058.x. [DOI] [PubMed] [Google Scholar]
  19. Geha R. S., Schneeberger E., Rosen F. S., Merler E. Interaction of human thymus-derived and non-thymus-derived lymphocytes in vitro. Induction of proliferation and antibody synthesis in B lymphocytes by a soluble factor released from antigen-stimulated T lymphocytes. J Exp Med. 1973 Nov 1;138(5):1230–1247. doi: 10.1084/jem.138.5.1230. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Gisler R. H., Staber F., Rüde E., Dukor P. Soluble mediators of T-B interaction. Eur J Immunol. 1973 Oct;3(10):650–652. doi: 10.1002/eji.1830031012. [DOI] [PubMed] [Google Scholar]
  21. Gorczynski R. M., Miller R. G., Phillips R. A. Reconstitution of T cell-depleted spleen cell populations by factors derived from T cells. 3. Mechanism of action of T cell-derived factors. J Immunol. 1973 Sep;111(3):900–913. [PubMed] [Google Scholar]
  22. HASKINS W. T., LANDY M., MILNER K. C., RIBI E. Biological properties of parent endotoxins and lipoid fractions, with a kinetic study of acid-hydrolyzed endotoxin. J Exp Med. 1961 Nov 1;114:665–684. doi: 10.1084/jem.114.5.665. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Hünig T., Schimpl A., Wecker E. Autoradiographic studies on the proliferation of antibody-producing cells in vitro. J Exp Med. 1974 Mar 1;139(3):754–760. doi: 10.1084/jem.139.3.754. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Katz D. H., Benacerraf B. The regulatory influence of activated T cells on B cell responses to antigen. Adv Immunol. 1972;15:1–94. doi: 10.1016/s0065-2776(08)60683-5. [DOI] [PubMed] [Google Scholar]
  25. Kessel R. W., Freedman H. H., Braun W. Relation of polysaccharide content to some biological properties of endotoxins from mutants of Salmonella typhimurium. J Bacteriol. 1966 Sep;92(3):592–596. doi: 10.1128/jb.92.3.592-596.1966. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Kolb C., Di Pauli R., Weiler E. Induction of IgG by lipid A in the newborn mouse. J Exp Med. 1974 Mar 1;139(3):467–478. doi: 10.1084/jem.139.3.467. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Leong D. L., Rudbach J. A. Antigenic competition between and endotoxic adjuvant and a protein antigen. Infect Immun. 1971 Feb;3(2):308–317. doi: 10.1128/iai.3.2.308-317.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Louis J. A., Chiller J. M., Weigle W. O. The ability of bacterial lipopolysaccharide to modulate the induction of unresponsiveness to a state of immunity. Cellular parameters. J Exp Med. 1973 Dec 1;138(6):1481–1495. doi: 10.1084/jem.138.6.1481. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. MILNER K. C., ANACKER R. L., FUKUSHI K., HASKINS W. T., LANDY M., MALMGREN B., RIBI E. SYMPOSIUM ON RELATIONSHIP OF STRUCTURE OF MICROORGANISMS TO THEIR IMMUNOLOGICAL PROPERTIES. III. STRUCTURE AND BIOLOGICAL PROPERTIES OF SURFACE ANTIGENS FROM GRAM-NEGATIVE BACTERIA. Bacteriol Rev. 1963 Dec;27:352–368. doi: 10.1128/br.27.4.352-368.1963. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. OSBORN M. J., ROSEN S. M., ROTHFIELD L., ZELEZNICK L. D., HORECKER B. L. LIPOPOLYSACCHARIDE OF THE GRAM-NEGATIVE CELL WALL. Science. 1964 Aug 21;145(3634):783–789. doi: 10.1126/science.145.3634.783. [DOI] [PubMed] [Google Scholar]
  31. Peavy D. L., Shands J. W., Jr, Adler W. H., Smith R. T. Mitogenicity of bacterial endotoxins: characterization of the mitogenic principle. J Immunol. 1973 Aug;111(2):352–357. [PubMed] [Google Scholar]
  32. Rubin A. S., Coons A. H. Specific heterologous enhancement of immune responses. 3. Partial characterization of supernatant material with enhancing activity. J Immunol. 1972 Jun;108(6):1597–1604. [PubMed] [Google Scholar]
  33. Rubin A. S., Coons A. H. Specific heterologous enhancement of immune responses. IV. Specific generation of a thymus-derived enhancing factor. J Exp Med. 1972 Dec 1;136(6):1501–1517. doi: 10.1084/jem.136.6.1501. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Rudbach J. A., Anacker R. L., Haskins W. T., Milner K. C., Ribi E. Physical structure of a native protoplasmic polysaccharide from Escherichia coli. J Immunol. 1967 Jan;98(1):1–7. [PubMed] [Google Scholar]
  35. Rudbach J. A. Molecular immunogenicity of bacterial lipopolysaccharide antigens: establishing a quantitative system. J Immunol. 1971 Apr;106(4):993–1001. [PubMed] [Google Scholar]
  36. Schimpl A., Wecker E. Replacement of T-cell function by a T-cell product. Nat New Biol. 1972 May 3;237(70):15–17. doi: 10.1038/newbio237015a0. [DOI] [PubMed] [Google Scholar]
  37. Schimpl A., Wecker E. Stimulation of IgG antibody response in vitro by T cell-replacing factor. J Exp Med. 1973 Feb 1;137(2):547–552. doi: 10.1084/jem.137.2.547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  38. Schmidtke J. R., Dixon F. J. Immune response to a hapten coupled to a nonimmunogenic carrier. Influence of lipopolysaccharide. J Exp Med. 1972 Aug 1;136(2):392–397. doi: 10.1084/jem.136.2.392. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Schrader J. W. Specific activation of the bone marrow-derived lymphocyte by antigen presented in a non-multivalent form. J Exp Med. 1973 Mar 1;137(3):844–849. doi: 10.1084/jem.137.3.844. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Sjöberg O., Andersson J., Möller G. Lipopolysaccharide can substitute for helper cells in the antibody response in vitro. Eur J Immunol. 1972 Aug;2(4):326–331. doi: 10.1002/eji.1830020406. [DOI] [PubMed] [Google Scholar]
  41. Sjöberg O., Andersson J., Möller G. Reconstitution of the antibody response in vitro of T cell-deprived spleen cells by supernatants from spleen cell cultures. J Immunol. 1972 Dec;109(6):1379–1385. [PubMed] [Google Scholar]
  42. Watson J. The role of humoral factors in the initiation of in vitro primary immune responses. 3. Characterization of factors that replace thymus-derived cells. J Immunol. 1973 Nov;111(5):1301–1313. [PubMed] [Google Scholar]
  43. Watson J., Trenkner E., Cohn M. The use of bacterial lipopolysaccharides to show that two signals are required for the induction of antibody synthesis. J Exp Med. 1973 Sep 1;138(3):699–714. doi: 10.1084/jem.138.3.699. [DOI] [PMC free article] [PubMed] [Google Scholar]

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