Skip to main content
NCBI home page
The Journal of Experimental Medicine logoLink to The Journal of Experimental Medicine
. 1978 Mar 1;147(3):800–813. doi: 10.1084/jem.147.3.800

Immunologic responsiveness of the C3H/HeJ mouse: differential ability of butanol-extracted lipopolysaccharide (LPS) to evoke LPS-mediated effects

PMCID: PMC2184202  PMID: 75941

Abstract

The lipopolysaccharide (LPS)-protein complex extracted from the cell wall of Escherichia coli K235 by the butanol-water technique has been shown to evoke a mitogenic response in bone marrow-derived (B) lymphocytes from the C3H/HeJ mouse strain. These mice are resistant to the effects of LPS extracted with phenol. Therefore, the ability of butanol-extracted LPS to modulate a spectrum of C3H/HeJ B-cell functions was investigated. Both butanol-extracted (LPS-B) and phenol- extracted (LPS-P) LPS preparations activated responder C3H/St spleen cell cultures to polyclonal antibody production, while only LPS-B activated C3H/HeJ spleen cells. Both LPS-P and LPS-B acted as adjuvants when injected after aggregated human gamma globulin (HGG) in C3H/St mice, but neither preparation was effective as a adjuvant in C3H/HeJ mice. LPS-P injected with deaggregated HGG (tolerogen) into LPS- sensitive mice has been shown previously to inhibit the induction of tolerance HGG. In the present studies, it was shown that LPS-B, but not LPs-p, was able to inhibit tolerance induction to HGG in the C3H/HeJ, whereas both preparations were effective in the C3H/St. LPS has also been shown to bypass tolerant T cells in LPS-sensitive mice late in tolerance to HGG at a time when B cells are responsive. However, in the C3H/HeJ, neither LPS-B nor LPS-P was capable of this function. The responsiveness of these B cells to HGG was demonstrated in transfer experiments. Thus, in the C3H/HeJ, LPS-B stimulates mitogenesis, polyclonal B-cell activation, and inhibition of tolerance induction, but cannot act as an effective adjuvant or as a bypass mechanism to activate B cells in the presence of tolerant T cells. The explanation for this pattern of responses may be attributable to yet another cellular defect in the C3H/HeJ mouse.

Full Text

The Full Text of this article is available as a PDF (955.1 KB).

Selected References

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

  1. Andersson J., Möller G., Sjöberg O. Selective induction of DNA synthesis in T and B lymphocytes. Cell Immunol. 1972 Aug;4(4):381–393. doi: 10.1016/0008-8749(72)90040-8. [DOI] [PubMed] [Google Scholar]
  2. 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]
  3. Andersson J., Sjöberg O., Möller G. Mitogens as probes for immunocyte activation and cellular cooperation. Transplant Rev. 1972;11:131–177. doi: 10.1111/j.1600-065x.1972.tb00048.x. [DOI] [PubMed] [Google Scholar]
  4. Armerding D., Katz D. H. Activation of T and B lymphocytes in vitro. I. Regulatory influence of bacterial lipopolysaccharide (LPS) on specific T-cell helper function. J Exp Med. 1974 Jan 1;139(1):24–43. doi: 10.1084/jem.139.1.24. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. BENNETT I. L., Jr, CLUFF L. E. Bacterial pyrogens. Pharmacol Rev. 1957 Dec;9(4):427–479. [PubMed] [Google Scholar]
  6. Bennett W. E., Cohn Z. A. The isolation and selected properties of blood monocytes. J Exp Med. 1966 Jan 1;123(1):145–160. doi: 10.1084/jem.123.1.145. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. CLAMAN H. N. TOLERANCE TO A PROTEIN ANTIGEN IN ADULT MICE AND THE EFFECT OF NONSPECIFIC FACTORS. J Immunol. 1963 Dec;91:833–839. [PubMed] [Google Scholar]
  8. Chedid L., Parant M., Damais C., Parant F., Juy D., Galelli A. Failure of endotoxin to increase nonspecific resistance to infection of lipopolysaccharide low-responder mice. Infect Immun. 1976 Mar;13(3):722–727. doi: 10.1128/iai.13.3.722-727.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. 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]
  10. Chiller J. M., Weigle W. O. Cellular events during induction of immunologic unresponsiveness in adult mice. J Immunol. 1971 Jun;106(6):1647–1653. [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. DES PREZ R. M., HOROWITZ H. I., HOOK E. W. Effects of bacterial endotoxin on rabbit platelets. I. Platelet aggregation and release of platelet factors in vitro. J Exp Med. 1961 Dec 1;114:857–874. doi: 10.1084/jem.114.6.857. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. DRESSER D. W. Effectiveness of lipid and lipidophilic substances as adjuvants. Nature. 1961 Sep 16;191:1169–1171. doi: 10.1038/1911169a0. [DOI] [PubMed] [Google Scholar]
  14. Diener E., Kraft N., Lee K. C., Shiozawa C. Antigen recognition. IV. Discrimination by antigen-binding immunocompetent B cells between immunity and tolerance is determined by adherent cells. J Exp Med. 1976 Apr 1;143(4):805–821. doi: 10.1084/jem.143.4.805. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Doyle M. V., Parks D. E., Weigle W. O. Specific suppression of the immune response by HGG tolerant spleen cells. I. Parameters affecting the level of suppression. J Immunol. 1976 Jun;116(6):1640–1645. [PubMed] [Google Scholar]
  16. Gery I., Krüger J., Spiesel S. Z. Stimulation of B-lymphocytes by endotoxin. Reactions of thymus-deprived mice and karyotypic analysis of dividing cells in mice bearing T 6 T 6 thymus grafts. J Immunol. 1972 Apr;108(4):1088–1091. [PubMed] [Google Scholar]
  17. Golub E. S., Mishell R. I., Weigle W. O., Dutton R. W. A modification of the hemolytic plaque assay for use with protein antigens. J Immunol. 1968 Jan;100(1):133–137. [PubMed] [Google Scholar]
  18. Golub E. S., Weigle W. O. Studies on the induction of immunologic unresponsiveness. I. Effects of endotoxin and phytochemagglutinin. J Immunol. 1967 Jun;98(6):1241–1247. [PubMed] [Google Scholar]
  19. 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]
  20. Goodman M. G., Weigle W. O. Nonspecific activation of murine lymphocytes. I. Proliferation and polyclonal activation induced by 2-mercaptoethanol and alpha-thioglycerol. J Exp Med. 1977 Mar 1;145(3):473–489. doi: 10.1084/jem.145.3.473. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Hamaoka T., Katz D. H. Cellular site of action of various adjuvants in antibody responses to hapten-carrier conjugates. J Immunol. 1973 Nov;111(5):1554–1563. [PubMed] [Google Scholar]
  22. Hartmann K., Dutton R. W., McCarthy M. M., Mishell R. I. Cell components in the immune response. II. Cell attachment separation of immune cells. Cell Immunol. 1970 Jul;1(2):182–189. doi: 10.1016/0008-8749(70)90005-5. [DOI] [PubMed] [Google Scholar]
  23. Jacobs D. M., Morrison D. C. Inhibition of the mitogenic response to lipopolysaccharide (LPS) in mouse spleen cells by polymyxin B. J Immunol. 1977 Jan;118(1):21–27. [PubMed] [Google Scholar]
  24. Jacobs M. D., Morrison D. C. Dissociation between mitogenicity and immunogenicity of TNP-lipopolysaccharide, a T-independent antigen. J Exp Med. 1975 Jun 1;141(6):1453–1458. doi: 10.1084/jem.141.6.1453. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Jerne N. K., Nordin A. A. Plaque Formation in Agar by Single Antibody-Producing Cells. Science. 1963 Apr 26;140(3565):405–405. doi: 10.1126/science.140.3565.405. [DOI] [PubMed] [Google Scholar]
  26. Lemke H., Coutinho A., Opitz H. G., Gronowicz E. Macrophages suppress direct B-cell activation by lipopolysaccharide. Scand J Immunol. 1975;4(7):707–720. doi: 10.1111/j.1365-3083.1975.tb02679.x. [DOI] [PubMed] [Google Scholar]
  27. 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]
  28. McIntire F. C., Sievert H. W., Barlow G. H., Finley R. A., Lee A. Y. Chemical, physical, biological properties of a lipopolysaccharide from Escherichia coli K-235. Biochemistry. 1967 Aug;6(8):2363–2372. doi: 10.1021/bi00860a011. [DOI] [PubMed] [Google Scholar]
  29. Melchers F., Andersson J. Synthesis, surface deposition and secretion of immunoglobulin M in bone marrow-derived lymphocytes before and after mitogenic stimulation. Transplant Rev. 1973;14:76–130. doi: 10.1111/j.1600-065x.1973.tb00103.x. [DOI] [PubMed] [Google Scholar]
  30. Mishell R. I., Dutton R. W. Immunization of dissociated spleen cell cultures from normal mice. J Exp Med. 1967 Sep 1;126(3):423–442. doi: 10.1084/jem.126.3.423. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. 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]
  32. 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]
  33. Ness D. B., Smith S., Talcott J. A., Grumet F. C. T cell requirements for the expression of the lipopolysaccharide adjuvant effect in vivo: evidence for a T cell-dependent and a T cell-independent mode of action. Eur J Immunol. 1976 Sep;6(9):650–654. doi: 10.1002/eji.1830060911. [DOI] [PubMed] [Google Scholar]
  34. Parks D. E., Doyle M. V., Weigle W. O. Effect of lipopolysaccharide on immunogenicity and tolerogenicity of HGG in C57BL/6J nude mice: evidence for a possible B cell deficiency. J Immunol. 1977 Dec;119(6):1923–1932. [PubMed] [Google Scholar]
  35. Persson U., Möller E. The effect of lipopolysaccharide on the primary immune response to the hapten NNP. Scand J Immunol. 1975 Sep;4(5-6):571–581. doi: 10.1111/j.1365-3083.1975.tb02663.x. [DOI] [PubMed] [Google Scholar]
  36. Persson U., Möller G. Effect of polyclonal B-cell activators on DNA synthesis in fibroblasts. Scand J Immunol. 1975 Sep;4(5-6):527–534. doi: 10.1111/j.1365-3083.1975.tb02658.x. [DOI] [PubMed] [Google Scholar]
  37. Poe W. J., Michael J. G. Separation of the mitogenic and antigenic responses to bacterial lipopolysaccharide. Immunology. 1976 Feb;30(2):241–248. [PMC free article] [PubMed] [Google Scholar]
  38. Schrader J. W. Induction of immunological tolerance to a thymus-dependent antigen in the absence of thymus-derived cells. J Exp Med. 1974 May 1;139(5):1303–1316. doi: 10.1084/jem.139.5.1303. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Shinohara N., Kern M. Differentiation of lymphoid cells: B cell as a direct target and T cell as a regulator in lipopolysaccharide-enhanced induction of immunoglobulin production. J Immunol. 1976 Jun;116(6):1607–1612. [PubMed] [Google Scholar]
  40. 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]
  41. Skidmore B. J., Chiller J. M., Weigle W. O. Immunologic properties of bacterial lipopolysaccharide (LPS). IV. Cellular basis of the unresponsiveness of C3H/HeJ mouse spleen cells to LPS-induced mitogenesis. J Immunol. 1977 Jan;118(1):274–281. [PubMed] [Google Scholar]
  42. Skidmore B. J., Chiller J. M., Weigle W. O., Riblet R., Watson J. Immunologic properties of bacterial lipopolysaccharide (LPS). III. Genetic linkage between the in vitro mitogenic and in vivo adjuvant properties of LPS. J Exp Med. 1976 Jan 1;143(1):143–150. doi: 10.1084/jem.143.1.143. [DOI] [PMC free article] [PubMed] [Google Scholar]
  43. Skidmore B. J., Morrison D. C., Chiller J. M., Weigle W. O. Immunologic properties of bacterial lipopolysaccharide (LPS). II. The unresponsiveness of C3H/HeJ Mouse spleen cells to LPS-induced mitogenesis is dependent on the method used to extract LPS. J Exp Med. 1975 Dec 1;142(6):1488–1508. doi: 10.1084/jem.142.6.1488. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. Smith S. M., Snyder I. S. Effect of lipopolysaccharide and lipid A on mouse liver pyruvate kinase activity. Infect Immun. 1975 Nov;12(5):993–998. doi: 10.1128/iai.12.5.993-998.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  45. Spitznagel J. K., Allison A. C. Mode of action of adjuvants: effects on antibody responses to macrophage-associated bovine serum albumin. J Immunol. 1970 Jan;104(1):128–139. [PubMed] [Google Scholar]
  46. Sultzer B. M. Genetic control of host responses to endotoxin. Infect Immun. 1972 Jan;5(1):107–113. doi: 10.1128/iai.5.1.107-113.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  47. Sultzer B. M. Genetic control of leucocyte responses to endotoxin. Nature. 1968 Sep 21;219(5160):1253–1254. doi: 10.1038/2191253a0. [DOI] [PubMed] [Google Scholar]
  48. Sultzer B. M. Genetic factors in leucocyte responses to endotoxin: further studies in mice. J Immunol. 1969 Jul;103(1):32–38. [PubMed] [Google Scholar]
  49. 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]
  50. Watson J., Riblet R. Genetic control of responses to bacterial lipopolysaccharides in mice. I. Evidence for a single gene that influences mitogenic and immunogenic respones to lipopolysaccharides. J Exp Med. 1974 Nov 1;140(5):1147–1161. doi: 10.1084/jem.140.5.1147. [DOI] [PMC free article] [PubMed] [Google Scholar]
  51. 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]
  52. Yoshinaga M., Yoshinaga A., Waksman B. H. Regulation of lymphocyte responses in vitro. I. Regulatory effect of macrophages and thymus-dependent (T) cells on the response of thymus-independent (B) lymphocytes to endotoxin. J Exp Med. 1972 Oct 1;136(4):956–961. doi: 10.1084/jem.136.4.956. [DOI] [PMC free article] [PubMed] [Google Scholar]

Articles from The Journal of Experimental Medicine are provided here courtesy of The Rockefeller University Press

RESOURCES