TLR9-Dependent and Independent Pathways Drive Activation of the Immune System by Propionibacterium Acnes

Propionibacterium acnes is usually a relatively harmless commensal. However, under certain, poorly understood conditions it is implicated in the etiology of specific inflammatory diseases. In mice, P. acnes exhibits strong immunomodulatory activity leading to splenomegaly, intrahepatic granuloma formation, hypersensitivity to TLR ligands and endogenous cytokines, and enhanced resistance to infection. All these activities reach a maximum one week after P. acnes priming and require IFN-γ and TLR9. We report here the existence of a markedly delayed (1–2 weeks), but phenotypically similar TLR9-independent immunomodulatory response to P. acnes. This alternative immunomodulation is also IFN-γ dependent and requires functional MyD88. From our experiments, a role for MyD88 in the IFN-γ-mediated P. acnes effects seems unlikely and the participation of the known MyD88-dependent receptors, including TLR5, Unc93B-dependent TLRs, IL-1R and IL-18R in the development of the alternative response has been excluded. However, the crucial role of MyD88 can partly be attributed to TLR2 and TLR4 involvement. Either of these two TLRs, activated by bacteria and/or endogenously generated ligands, can fulfill the required function. Our findings hint at an innate immune sensitizing mechanism, which is potentially operative in both infectious and sterile inflammatory disorders

High susceptibility to lipopolysaccharide-induced lethal shock in encephalomyocarditis virus-infected mice

Secondary bacterial infection in humans is one of the pathological conditions requiring clinical attention. In this study, we examined the effect of lipopolysaccharide (LPS) on encephalomyocarditis virus (EMCV) infected mice. All mice inoculated with EMCV at 5 days before LPS challenge died within 24 h. LPS-induced TNF-α mRNA expression was significantly increased in the brain and heart at 5 days after EMCV infection. CD11b+/TLR4+ cell population in the heart was remarkably elevated at 5 days after EMCV infection, and sorted CD11b+ cells at 5 days after EMCV infection produced a large amount of TNF-α on LPS stimulation in vivo and in vitro. In conclusion, we found that the infiltration of CD11b+ cells into infected organs is involved in the subsequent LPS-induced lethal shock in viral encephalomyocarditis. This new experimental model can help define the mechanism by which secondary bacterial infection causes a lethal shock in viral encephalomyocarditis

A striking correlation between lethal activity and apoptotic DNA fragmentation of liver in response of D-galactosamine- sensitized mice to a non-lethal amount of lipopolysaccharide

AIM: To observe whether challenge of bacterial lipopolysaccharide (LPS) with D-galactosamine (D-GalN) in mice will result in apoptotic characteristic of vital organs. METHODS: The experimental group of mice was challenged directly with bacterial LPS (0.05 μg) in the presence of D-GalN (20 mg), and the control group of mice was challenged either with bacterial LPS or with D-GalN alone. The organs including brain, kidney, heart, spleen, lung, and liver were removed at an indicated time point under ether anethesia, and immediately homogenized, from which DNA was extracted. The DNA obtained from these organs was analyzed by agarose gel electrophoresis to determine whether the DNA laddering phenomenon existed. The amount of plasma LDH and GOT was detected in mice challenged with bacterial LPS in the presence of D-GalN, and either bacterial LPS or D-GalN alone. RESULTS: Apoptotic DNA fragmentation was initially seen at 4 h after challenge only in the livers of mice challenged with bacterial LPS and D-GalN, all mice in this group challenged with bacterial LPS and D-GalN died at 7 h after challenge; in contrast, the animals in the control group were all alive and the DNA was integral. CONCLUSION: The liver is the only specific target organ where apoptotic DNA fragmentation was present in mice treated with D- GalN and challenged with bacterial LPS and the liver impairment might be the critical cause of the lethality of mice elicited by bacterial LPS

Lethal effect and apoptotic DNA fragmentation in response of D-GalN-treated mice to bacterial LPS can be suppressed by pre-exposure to minute amount of bacterial LPS: Dual role of TNF receptor 1

AIM: To investigate whether induction of tolerance of mice to lipopolysaccharide (LPS) was able to inhibit apoptotic reaction in terms of characteristic DNA fragmentation and protect mice from lethal effect. METHODS: Experimental groups of mice were pretreated with non-lethal amount of LPS (0.05 μg). Both control and experimental groups simultaneously were challenged with LPS plus D-GalN for 6-7 h. The evaluations of both DNA fragmentations from the livers and the protection efficacy against lethality to mice through induction of tolerance to LPS were conducted. RESULTS: In the naive mice challenge with LPS plus D-GalN resulted in complete death in 24 h, whereas a characteristic apoptotic DNA fragmentation was exclusively seen in the livers of mice receiving LPS in combination with D-GalN. The mortality in the affected mice was closely correlated to the onset of DNA fragmentation. By contrast, in the mice pre-exposed to LPS, both lethal effect and apoptotic DNA fragmentation were suppressed when challenged with LPS/D-GalN. In addition to LPS, the induction of mouse tolerance to TNF also enabled mice to cross-react against death and apoptotic DNA fragmentation when challenged with TNF and/or LPS in the presence of D-GalN. Moreover, this protection effect by LPS could last up to 24 h. TNFR1 rather than TNFR2 played a dual role in signaling pathway of either induction of tolerance to LPS for the protection of mice from mortality or inducing morbidity leading to the death of mice. CONCLUSION: The mortality of D-GalN-treated mice in response to LPS was exceedingly correlated to the onset of apoptosis in the liver, which can be effectively suppressed by brief exposure of mice to a minute amount of LPS. The induced tolerance status was mediated not only by LPS but also by TNF. The developed tolerance to either LPS or TNF can be reciprocally cross-reacted between LPS and TNF challenges, whereas the signaling of induction of tolerance and promotion of apoptosis was through TNFR1, rather than TNFR2

Stage of primary infection with lymphocytic choriomeningitis virus determines predisposition or resistance of mice to secondary bacterial infections

We investigated the effect of a primary non-lethal infection with lymphocytic choriomeningitis virus (LCMV) on the course and outcome of a secondary infection with the Gram-negative Salmonella enterica serovar Typhimurium or the Gram-positive Listeria monocytogenes in mice. We found that at each stage of the viral infection the susceptibility of mice to bacterial super-infections changes dramatically and depends also on whether the secondary infection is a Gram-positive or Gram-negative one. The study shows that the outcome of the secondary infection is determined by a delicate balance between the overproduction of and the hypersensitivity to inflammatory cytokines (TNF-α and IFN-γ), as well as by the changes in blood leukocytes occurring in mice in the course of viral infection

Role of lipopolysaccharide susceptibility in the innate immune response to Salmonella typhimurium infection: LPS, a primary target for recognition of Gram-negative bacteria

Lipopolysaccharide is an important recognition marker by virtue of which the innate immune system senses and reacts against Gram-negative bacteria invading the LPS susceptible host. This review deals with the factors affecting LPS susceptibility and with the role of the latter in the course and outcome of Salmonella typhimurium infection. (C) 2001 Editions scientifiques et medicales Elsevier SAS

Role of interferons in LPS hypersensitivity

The innate immune response to Gram-negative bacteria depends mainly on the ability of the host to respond to the LPS component. Consequently, the state of LPS sensitivity at the time of infection and the numbers of invading bacteria (i.e. the amounts of LPS) are primary factors determining the innate responses provoked by Gram-negative pathogens. LPS sensitivity increases following treatment of mice with live or killed micro-organisms. Two types of sensitization have been recognized, strong, IFN-γ-dependent and moderate IFN-γ-independent. IL-12 and IL-18 are intimately involved in the induction of IFN-γ by bacteria. We showed that Gram-negative bacteria induce IFN-γ in mice also by an IFN-β-dependent pathway that requires IL-18 and is independent of IL-12 signaling. This pathway is STAT4 dependent, the activation of which is directly linked to IFN-β. Further, IFN-β can be replaced by IFN-α. While different components of Gram-negative bacteria induce IL-12 and IL-18, LPS seems to be the only component in these bacteria capable of inducing IFN-β. Therefore, the IFN-β pathway of IFN-γ induction, unlike the IL-12 pathway, proceeds only in LPS responder mice. The IFN-α/β-dependent pathway is expected to play a role whenever IFN-α or IFN-β, and IL-18 are produced concomitantly during infection