A comparison of T cell memory against the same antigen induced by virus versus intracellular bacteria.
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Cytotoxic T cell (CTL) memory was analyzed after infection with lymphocytic choriomeningitis virus (LCMV) and recombinant Listeria monocytogenes (rLM) expressing the complete nucleoprotein of LCMV (rLM-NP(actA)) or only the immunodominant epitope of H-2(d) mice (rLM-NP(118-126)). Immunization with LCMV and rLM induced a long-lived increased CTL precursor (CTLp) frequency specific for the viral (NP(118-126)) and for the bacterial (LLO(91-99)) epitope, respectively. However, after infection with rLM memory, CTLs were less protective against an intravenous LCMV challenge infection than a comparable number of LCMV-induced memory T cells. LCMV, but not recombinant Listeria-induced memory T cells were able to protect against lethal choriomeningitis by LCMV or a subsequent peripheral infection with recombinant vaccinia virus expressing LCMV-NP. The protective memory after viral and after rLM immunization was paralleled by evidence of LCMV but not rLM antigen persistence on day 15 and 30 after vaccination. These results document a striking difference in protective T cell memory between viral and bacterial vaccines and indicate that rapid T cell-dependent immune protection correlates with antigen persistence. (+info)
H2-M3-restricted T cells in bacterial infection: rapid primary but diminished memory responses.
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Major histocompatibility complex (MHC) class Ib molecules have been implicated in CD8(+) T cell-mediated defenses against intracellular bacterial infection, but the relative importance of MHC class Ib-restricted T cells in antimicrobial immunity is unknown. In this report, we use MHC tetramers to characterize T cell responses restricted by H2-M3, an MHC class Ib molecule that selectively presents N-formyl peptides. We find that sizeable H2-M3-restricted T cell responses, occurring earlier than MHC class Ia-restricted T cell responses, are mounted after primary infection with the intracellular bacterium Listeria monocytogenes. These H2-M3-restricted T cells are cytolytic and produce interferon gamma. However, after a second L. monocytogenes infection, H2-M3-restricted memory T cell responses are minor in comparison to the much larger MHC class Ia-restricted responses. This first direct characterization of an MHC class Ib-restricted T cell response indicates that CD8(+) T cells responding to L. monocytogenes infection can be divided into two groups: H2-M3-restricted responses, which provide rapid and quantitatively substantial effector function during primary infections but contribute relatively little to memory responses, and MHC class Ia-restricted responses, which expand later during primary infection but form memory T cells that respond rapidly and dramatically in response to subsequent infections by the same pathogen. (+info)
The bvr locus of Listeria monocytogenes mediates virulence gene repression by beta-glucosides.
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The beta-glucoside cellobiose has been reported to specifically repress the PrfA-dependent virulence genes hly and plcA in Listeria monocytogenes NCTC 7973. This led to the hypothesis that beta-glucosides, sugars of plant origin, may act as signal molecules, preventing the expression of virulence genes if L. monocytogenes is living in its natural habitat (soil). In three other laboratory strains (EGD, L028, and 10403S), however, the effect of cellobiose was not unique, and all fermentable carbohydrates repressed hly. This suggested that the downregulation of virulence genes by beta-glucosides is not a specific phenomenon but, rather, an aspect of a global regulatory mechanism of catabolite repression (CR). We assessed the effect of carbohydrates on virulence gene expression in a panel of wild-type isolates of L. monocytogenes by using the PrfA-dependent phospholipase C gene plcB as a reporter. Utilization of any fermentable sugar caused plcB repression in wild-type L. monocytogenes. However, an EGD variant was identified in which, as in NCTC 7973, plcB was only repressed by beta-glucosides. Thus, the regulation of L. monocytogenes virulence genes by sugars appears to be mediated by two separate mechanisms, one presumably involving a CR pathway and another specifically responding to beta-glucosides. We have identified in L. monocytogenes a 4-kb operon, bvrABC, encoding an antiterminator of the BglG family (bvrA), a beta-glucoside-specific enzyme II permease component of the phosphoenolpyruvate-sugar phosphotransferase system (bvrB), and a putative ADP-ribosylglycohydrolase (bvrC). Low-stringency Southern blots showed that this locus is absent from other Listeria spp. Transcription of bvrB was induced by cellobiose and salicin but not by arbutin. Disruption of the bvr operon by replacing part of bvrAB with an interposon abolished the repression by cellobiose and salicin but not that by arbutin. Our data indicate that the bvr locus encodes a beta-glucoside-specific sensor that mediates virulence gene repression upon detection of cellobiose and salicin. Bvr is the first sensory system found in L. monocytogenes that is involved in environmental regulation of virulence genes. (+info)
Isolation of rifampin-resistant mutants of Listeria monocytogenes and their characterization by rpoB gene sequencing, temperature sensitivity for growth, and interaction with an epithelial cell line.
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The sequence of the rpoB gene from Listeria monocytogenes was determined. Rifampin-resistant (Rif(r)) mutants arising from L. monocytogenes cultures exposed to rifampin were isolated, and by partial sequencing of their rpoB genes, seven different point mutations affecting five different amino acids (473Asp-->Asn or Gly, 479Gly-->Asp, 483His-->Tyr or Leu, 528Ile-->Phe, and 530Ser-->Tyr), which led to MICs of 0.5 to 100 microg/ml for the organisms, were determined. These mutants showed various deficiencies for growth at 42 degrees C, with only one being comparable to the wild-type strain. The interaction of these Rif(r) mutants with human Caco-2 cells was examined by using an immunofluorescence technique. Three mutants failed to interact, while three showed a reduced interaction compared to that of the wild type. It is believed that these pleiotropic phenotypes have arisen as a result of mutations within the DNA-dependent RNA polymerase holoenzyme. (+info)
Role of IL-10 in a neonatal mouse listeriosis model.
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This study was undertaken to test the hypothesis that altered IL-10 production plays a role in the increased susceptibility of neonates to listeriosis. Plasma IL-10 levels were measured in neonatal and adult mice at various times after infection with Listeria monocytogenes. Relative to adults, neonatal mice had markedly increased IL-10 levels early in the course of infection with Listeria using a 90% lethal dose. Higher neonatal IL-10 responses were also observed after injecting adults and pups with equal doses of killed organisms. Splenic macrophages from neonates produced higher IL-10 levels than those of adults after in vitro stimulation with killed bacteria, confirming in vivo observations. Moreover, IL-10 blockade had differential effects in neonates and adults infected with live Listeria. In adult mice, anti-IL-10 Abs decreased bacterial burden early in the course of infection, but were no longer effective at 6 days or later after challenge. In the pups, however, the same treatment had beneficial effects both early and late during infection and resulted in increased survival. Collectively, our data suggest that an overproduction of IL-10 by macrophages may at least partially explain the increased susceptibility of neonates to listeriosis, and provide further evidence that cytokine production is different in adults and neonates. (+info)
Typing Listeria monocytogenes by random amplified polymorphic DNA (RAPD) fingerprinting.
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Twenty epidemiologically unrelated Listeria monocytogenes strains isolated from different animals, locations and on different dates in Japan were classified into 18 types by the random amplified polymorphic DNA (RAPD) fingerprinting technique with four primers. Further, seven epidemiologically related L. monocytogenes strains isolated from raw milk and a bulk tank on a dairy farm represented the same RAPD type suggesting that they were all of the same origin. Therefore, RAPD-polymerase chain reaction (PCR) analysis, which is rapid, simple and inexpensive to perform, can be used in surveys as a convenient epidemiological technique. (+info)
Sulfasalazine prevents T-helper 1 immune response by suppressing interleukin-12 production in macrophages.
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Interleukin-12 (IL-12) plays a pivotal role in the development of T-helper 1 (Th1) immune response, which may be involved in the pathogenesis of chronic inflammatory autoimmune disorders. In this study we investigated the effects of sulfasalazine, a drug for treating inflammatory bowel disease and rheumatoid arthritis, on the production of IL-12 from mouse macrophages stimulated with lipopolysaccharide (LPS). Sulfasalazine potently inhibited the production of IL-12 in a dose-dependent manner, in part through the down-regulation of nuclear factor kappaB (NFkappaB) activation in IL-12 p40 gene. Activation of macrophages by LPS resulted in markedly enhanced binding activities to the kappaB site, which significantly decreased upon addition of sulfasalazine as demonstrated by an electrophoretic gel shift assay. Importantly, macrophages pretreated with sulfasalazine either in vitro or in vivo reduced their ability to induce interferon-gamma (IFN-gamma) and increased the ability to induce IL-4 in antigen-primed CD4+ T cells. From these results, sulfasalazine may induce the Th2 cytokine profile in CD4+ T cells by suppressing IL-12 production in macrophages, and sulfasalazine-induced inhibition of IL-12 production in macrophages may explain some of the known biological effects of sulfasalazine. (+info)
The antibacterial efficacy of trovafloxacin against an experimental infection with Listeria monocytogenes in hydrocortisone-treated mice.
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The efficacy of trovafloxacin in treating Listeria monocytogenes infections in glucocorticosteroid-treated mice was compared with the efficacy of amoxycillin. Swiss mice were treated with daily injections of 2.5 mg hydrocortisone s.c. and then infected i.v. with 1 x 10(7) cfu of L. monocytogenes. Untreated, this level of infection resulted in 100% mortality between day 3 and day 5 after infection. Both s.c. trovafloxacin and amoxycillin were effective in reducing the number of viable L. monocytogenes in the liver and spleen. Although the MIC of amoxycillin for this isolate of L. monocytogenes was lower than that of trovafloxacin (0.063 mg/L versus 0.5 mg/L, respectively), trovafloxacin was more efficacious in vivo after a single dose in the dose range between 12.5 and 100 mg/kg than was amoxycillin. After treatment with trovafloxacin at 100 mg/kg bodyweight od for 3 days, a mean log10 cfu of 1.58 and 2.52 L. monocytogenes could be recovered from the spleens and livers, respectively, whereas after treatment with amoxycillin at 100 mg/kg bodyweight every 8 h for 3 days, the mean 1og10 cfu values were 2.36 and 2.02, respectively. These differences were statistically not significant. Results of the present study show that the antibacterial efficacy of trovafloxacin against L. monocytogenes in our animal model is equivalent to that of amoxycillin. (+info)