Quantitative aspects of lipopolysaccharide and cytokine requirements to generate nitric oxide in macrophages from LPS-hyporesponsive (Lps(d)) C3H/HeJ mice

• Published in: Folia microbiologica 2004-01, Vol.49 (6), p.737-744
• Main Authors: Kmonickova, E, Zidek, Z
• Format: Article
• Language: English
• Subjects: Animals; Biosynthesis; Cytokines; Cytokines - biosynthesis; Cytokines - immunology; Cytokines - metabolism; Escherichia coli - immunology; Escherichia coli - metabolism; Female; Interferon-gamma - immunology; Interferon-gamma - metabolism; Interleukin 10; Interleukin-10 - immunology; Interleukin-10 - metabolism; Lipopolysaccharides; Lipopolysaccharides - immunology; Macrophage Activation - immunology; Macrophages; Macrophages, Peritoneal - immunology; Macrophages, Peritoneal - metabolism; Medical research; Mice; Mice, Inbred C3H; Nitric oxide; Nitric Oxide - biosynthesis; Tumor Necrosis Factor-alpha - immunology; Tumor Necrosis Factor-alpha - metabolism; Tumor necrosis factor-α; γ-Interferon
• ISSN: 0015-5632; 1874-9356
• DOI: 10.1007/BF02931558

Due to a gene defect (Lps(d)), C3H/HeJ mice are known to be hyporesponsive to the immunobiological potential of lipopolysaccharide (LPS). We studied dose requirements for LPS, IFN-gamma, and cytokines TNF-alpha and IL-10 to produce nitric oxide (NO) in peritoneal macrophages (Mphi) from these animals. In contrast to the Lps(n) C3H/HeN mice, high concentrations of LPS (up to 5 microg/mL) or IFN-gamma (up to 5 ng/mL) by themselves were unable to activate NO production in C3H/HeJ Mphi. The failure to produce NO could not be overcome by addition of L-arginine or tetrahydropterin. The high-output NO biosynthesis was dose-dependently stimulated by combined administration of varying concentrations of IFN-gamma (50-5000 pg/mL) and LPS (approximately 1 ng/mL) or to a lesser extent by IFN-gamma plus TNF-alpha or TNF-alpha/IL-10. Formation of NO in C3H/HeJ MCO triggered by high concentration of LPS (approximately 1 microg/mL) given together with IFN-gamma (0.2-5 ng/mL) reached the values typical for Lps(n) C3H/HeN mice. While Mphi from C3H/HeN mice secreted TNF-alpha, IL-10, and IL-10 upon contact with a low dose of LPS (1 ng/mL), C3H/HeJ Mphi required high concentration of LPS (5 microg/mL) to enhance the secretion of the cytokines. Yet, this dose remained ineffective to stimulate IFN-gamma in Mphi from C3H/HeJ mice. It can be presumed that one of the important factors influencing their deficient ability to form NO is a failure of Mphi to produce IFN-gamma upon LPS contact.