Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells

T. Gutsmann, J. Howe, U. Z̈hringer, P. Garidel, A. B. Schromm, M. H J Koch, Yukari Fujimoto, K. Fukase, I. Moriyon, G. Martínez-De-Tejada, K. Brandenburg

Research output: Contribution to journalArticle

31 Citations (Scopus)

Abstract

The structural prerequisites for lipopolysaccharide (LPS) and its partial structures for the activation of the Limulus clotting cascade (Limulus amebocyte lysate [LAL] test) are described and compared with the corresponding requirements for the activation of human immune cells such as mononuclear cells. A necessary, but not sufficient, structural motif for this is the presence of the 4ĝ2-phosphate-diglucosamine backbone recognition structure (ĝepitopeĝ) in lipid A. High activity is only expressed by assemblies of endotoxins, but this is largely independent of the type of supramolecular aggregate structure. A particular conformation of the epitope within the lipid A assembly must be present, which is influenced by addition of further saccharide units to the lipid A moiety, but also reacts slightly to the acylation pattern. In contrast, the cytokine production of human immune cells induced by LPS sensitively depends on the type of its aggregate structure. In the case of a hexa-acylated bisphosphorylated lipid A structure, high activity is only observed with cubic inverted aggregates. Furthermore, addition of antimicrobial agents (such as polymyxin B) leads to a nearly complete inhibition of cytokine production, whereas the reduction in the Limulus assay is much lower. These data are important since a reliable determination of endotoxin concentrations, in particular with respect to its ability to elicit severe infections, is of high interest.

Original languageEnglish
Pages (from-to)39-47
Number of pages9
JournalInnate Immunity
Volume16
Issue number1
DOIs
Publication statusPublished - 2010 Feb
Externally publishedYes

Fingerprint

Limulus Test
Lipid A
Horseshoe Crabs
Cytokines
Endotoxins
Lipopolysaccharides
Epitopes
Polymyxin B
Acylation
Anti-Infective Agents
Phosphates
Infection

Keywords

  • Cytokine production
  • Endotoxic activity
  • Limulus test
  • Mononuclear cells
  • Structure

ASJC Scopus subject areas

  • Cell Biology
  • Molecular Biology
  • Immunology
  • Microbiology
  • Infectious Diseases

Cite this

Gutsmann, T., Howe, J., Z̈hringer, U., Garidel, P., Schromm, A. B., Koch, M. H. J., ... Brandenburg, K. (2010). Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells. Innate Immunity, 16(1), 39-47. https://doi.org/10.1177/1753425909106447

Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells. / Gutsmann, T.; Howe, J.; Z̈hringer, U.; Garidel, P.; Schromm, A. B.; Koch, M. H J; Fujimoto, Yukari; Fukase, K.; Moriyon, I.; Martínez-De-Tejada, G.; Brandenburg, K.

In: Innate Immunity, Vol. 16, No. 1, 02.2010, p. 39-47.

Research output: Contribution to journalArticle

Gutsmann, T, Howe, J, Z̈hringer, U, Garidel, P, Schromm, AB, Koch, MHJ, Fujimoto, Y, Fukase, K, Moriyon, I, Martínez-De-Tejada, G & Brandenburg, K 2010, 'Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells', Innate Immunity, vol. 16, no. 1, pp. 39-47. https://doi.org/10.1177/1753425909106447
Gutsmann, T. ; Howe, J. ; Z̈hringer, U. ; Garidel, P. ; Schromm, A. B. ; Koch, M. H J ; Fujimoto, Yukari ; Fukase, K. ; Moriyon, I. ; Martínez-De-Tejada, G. ; Brandenburg, K. / Structural prerequisites for endotoxic activity in the Limulus test as compared to cytokine production in mononuclear cells. In: Innate Immunity. 2010 ; Vol. 16, No. 1. pp. 39-47.
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