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. 1980 Jul;40(3):473–482.

Effect of lipid A-associated protein and lipid A on the expression of lipopolysaccharide activity. I. Immunological activity.

S Izui, D C Morrison, B Curry, F J Dixon
PMCID: PMC1458040  PMID: 7000681

Abstract

A detailed investigation has been made of the contribution of the various chemical moieties of bacterial endotoxins, namely lipid A-associated protein (LAP), lipid A and O-antigen polysaccharide to a number of the immunological activities of these active bacterial products. Advantage was taken of the availability of antigenically identical endotoxin preparations from Escherichia coli 0111:B4 which differed greatly in their content of LAP and/or lipid A. The capacity to initiate in vitro proliferative responses in murine splenocytes was in a large part related to the presence of LAP with a less potent, although still critical, dependence upon lipid A. On the other hand, the in vivo polyclonal antibody response was dependent only upon lipid A. In this respect, the presence of LAP had no apparent effect on the stimulation of nonspecific low affinity antibody. All preparations, regardless of LAP and lipid A content, stimulated similar in vivo enhancement of antibody responses to a protein antigen (adjuvanticity) and specific immune responses to the endotoxin polysaccharide antigen. The results emphasize the lack of correlation between in vitro B lymphocyte proliferative responses and in vivo immunostimulatory responses of bacterial endotoxin preparations. These data also suggest a minimal contribution of LAP to in vivo responses and an extremely limited contribution of lipid A to the adjuvant activity and the primary immune response to O-antigen polysaccharide.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. AMES B. N., DUBIN D. T. The role of polyamines in the neutralization of bacteriophage deoxyribonucleic acid. J Biol Chem. 1960 Mar;235:769–775. [PubMed] [Google Scholar]
  2. 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]
  3. Andersson J., Sjöberg O., Möller G. Induction of immunoglobulin and antibody synthesis in vitro by lipopolysaccharides. Eur J Immunol. 1972 Aug;2(4):349–353. doi: 10.1002/eji.1830020410. [DOI] [PubMed] [Google Scholar]
  4. Betz S. J., Morrison D. C. Chemical and biologic properties of a protein-rich fraction of bacterial lipopolysaccharides. I. The in vitro murine lymphocyte response. J Immunol. 1977 Oct;119(4):1475–1481. [PubMed] [Google Scholar]
  5. Britton S., Möller G. Regulation of antibody synthesis against Escherichia coli endotoxin. I. Suppressive effect of endogenously produced and passively transferred antibodies. J Immunol. 1968 Jun;100(6):1326–1334. [PubMed] [Google Scholar]
  6. CYNKIN M. A., ASHWELL G. Estimation of 3-deoxy sugars by means of the malonaldehyde-thiobarbituric acid reaction. Nature. 1960 Apr 9;186:155–156. doi: 10.1038/186155a0. [DOI] [PubMed] [Google Scholar]
  7. 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]
  8. Frank S., Specter S., Nowotny A., Friedman H. Immunocycte stimulation in vitro by nontoxic bacterial lipopolysaccharide derivatives. J Immunol. 1977 Sep;119(3):855–860. [PubMed] [Google Scholar]
  9. Goodman M. G., Morrison D. C., Weigle W. O. Modulation of lipopolysaccharide (LPS)-mediated function by structural differences of two physically distinct fractions of Escherichia coli K235 LPS. J Immunol. 1977 May;118(5):1852–1857. [PubMed] [Google Scholar]
  10. Goodman M. G., Parks D. E., Weigle W. O. Immunologic responsiveness of the C3H/HeJ mouse: differential ability of butanol-extracted lipopolysaccharide (LPS) to evoke LPS-mediated effects. J Exp Med. 1978 Mar 1;147(3):800–813. doi: 10.1084/jem.147.3.800. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hepper K. P., Garman R. D., Lyons M. F., Teresa G. W. Plaque-forming cell response in BALB/c mice to two preparations of LPS extracted from Salmonella enteritidis. J Immunol. 1979 Apr;122(4):1290–1293. [PubMed] [Google Scholar]
  12. Izui S., Eisenberg R. A., Dixon F. J. IgM rheumatoid factors in mice injected with bacterial lipopolysaccharides. J Immunol. 1979 May;122(5):2096–2102. [PubMed] [Google Scholar]
  13. Izui S., Kobayakawa T., Zryd M. J., Louis J., Lambert P. H. Mechanism for induction of anti-DNA antibodies by bacterial lipopolysaccharides in mice; II. Correlation between anti-DNA induction and polyclonal antibody formation by various polyclonal B lymphocyte activators. J Immunol. 1977 Dec;119(6):2157–2162. [PubMed] [Google Scholar]
  14. JOHNSON A. G., GAINES S., LANDY M. Studies on the O antigen of Salmonella typhosa. V. Enhancement of antibody response to protein antigens by the purified lipopolysaccharide. J Exp Med. 1956 Feb 1;103(2):225–246. doi: 10.1084/jem.103.2.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Kettman J., Dutton R. W. An in vitro primary immune response to 2,4,6-trinitrophenyl substituted erythrocytes: response against carrier and hapten. J Immunol. 1970 Jun;104(6):1558–1561. [PubMed] [Google Scholar]
  16. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  17. Landy M., Sanderson R. P., Jackson A. L. Humoral and cellular aspects of the immune response to the somatic antigen of Salmonella enteritidis. J Exp Med. 1965 Sep 1;122(3):483–504. doi: 10.1084/jem.122.3.483. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. McConahey P. J., Dixon F. J. A method of trace iodination of proteins for immunologic studies. Int Arch Allergy Appl Immunol. 1966;29(2):185–189. doi: 10.1159/000229699. [DOI] [PubMed] [Google Scholar]
  19. Morrison D. C., Betz S. J., Jacobs D. M. Isolation of a lipid A bound polypeptide responsible for "LPS-initiated" mitogenesis of C3H/HeJ spleen cells. J Exp Med. 1976 Sep 1;144(3):840–846. doi: 10.1084/jem.144.3.840. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Morrison D. C., Leive L. Fractions of lipopolysaccharide from Escherichia coli O111:B4 prepared by two extraction procedures. J Biol Chem. 1975 Apr 25;250(8):2911–2919. [PubMed] [Google Scholar]
  21. Morrison D. C., Ryan J. L. Bacterial endotoxins and host immune responses. Adv Immunol. 1979;28:293–450. doi: 10.1016/s0065-2776(08)60802-0. [DOI] [PubMed] [Google Scholar]
  22. Möller G. 19S antibody production against soluble lipopolysaccharide antigens by individual lymphoid cells in vitro. Nature. 1965 Sep 11;207(5002):1166–1168. doi: 10.1038/2071166a0. [DOI] [PubMed] [Google Scholar]
  23. Neter E. Endotoxins and the immune response. Curr Top Microbiol Immunol. 1969;47:82–124. doi: 10.1007/978-3-642-46160-6_5. [DOI] [PubMed] [Google Scholar]
  24. Rudbach J. A. Molecular immunogenicity of bacterial lipopolysaccharide antigens: establishing a quantitative system. J Immunol. 1971 Apr;106(4):993–1001. [PubMed] [Google Scholar]
  25. Skidmore B. J., Chiller J. M., Morrison D. C., Weigle W. O. Immunologic properties of bacterial lipopolysaccharide (LPS): correlation between the mitogenic, adjuvant, and immunogenic activities. J Immunol. 1975 Feb;114(2 Pt 2):770–775. [PubMed] [Google Scholar]
  26. Sultzer B. M., Goodman G. W. Endotoxin protein: a B-cell mitogen and polyclonal activator of C3H/HeJ lymphocytes. J Exp Med. 1976 Sep 1;144(3):821–827. doi: 10.1084/jem.144.3.821. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. VAUGHAN M., BERGER J. E., STEINBERG D. HORMONE-SENSITIVE LIPASE AND MONOGLYCERIDE LIPASE ACTIVITIES IN ADIPOSE TISSUE. J Biol Chem. 1964 Feb;239:401–409. [PubMed] [Google Scholar]
  28. Von Eschen K. B., Rudbach J. A. Antibody responses of mice to alkaline detoxifield lipopolysaccharide. J Immunol. 1976 Jan;116(1):8–11. [PubMed] [Google Scholar]
  29. Von Eschen K. B., Rudbach J. A. Immunological responses of mice to native protoplasmic polysaccharide and lipopolysaccharide: functional separation of the two signals required to stimulate a secondary antibody response. J Exp Med. 1974 Dec 1;140(6):1604–1614. doi: 10.1084/jem.140.6.1604. [DOI] [PMC free article] [PubMed] [Google Scholar]
  30. WARAVDEKAR V. S., SASLAW L. D. A sensitive colorimetric method for the estimation of 2-deoxy sugars with the use of the malonaldehyde-thiobarbituric acid reaction. J Biol Chem. 1959 Aug;234(8):1945–1950. [PubMed] [Google Scholar]

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