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Infection and Immunity May 2003Two pathogenic species in the genus Listeria, Listeria monocytogenes and Listeria ivanovii, are characterized by the production of hemolysins belonging to...
Differences in gamma interferon production induced by listeriolysin O and ivanolysin O result in different levels of protective immunity in mice infected with Listeria monocytogenes and Listeria ivanovii.
Two pathogenic species in the genus Listeria, Listeria monocytogenes and Listeria ivanovii, are characterized by the production of hemolysins belonging to cholesterol-dependent cytolysins, listeriolysin O (LLO) and ivanolysin O (ILO), respectively. LLO, produced by L. monocytogenes, is able to induce gamma interferon (IFN-gamma) production and contributes to the generation of Th1-dependent protective immunity. On the other hand, nothing is known about the role of ILO, produced by L. ivanovii, in this regard. In this study, we immunized mice with 0.1 50% lethal dose (LD(50)) of L. monocytogenes and L. ivanovii. Protective immunity against a challenge with 10 LD(50) was generated in mice infected with L. monocytogenes, whereas L. ivanovii infection did not induce protection. After immunization, the level of IFN-gamma in serum samples was increased in mice given L. monocytogenes but not in those given L. ivanovii. To determine the IFN-gamma-inducing activity of cytolysins, recombinant protein was constructed. Recombinant ILO exhibited significantly lower IFN-gamma-inducing activity than LLO. By comparing the IFN-gamma-inducing activity of a chimera incorporating LLO and ILO, it was found that domains 1 to 3 of LLO were critical for IFN-gamma-inducing activity while the counterpart in ILO was unable to induce cytokine production. These results suggested that the weak ability of ILO to induce IFN-gamma production is responsible for the failure of L. ivanovii to generate effective protective immunity.
Topics: Amino Acid Sequence; Animals; Bacterial Toxins; Female; Heat-Shock Proteins; Hemolysin Proteins; Interferon-gamma; Listeria; Listeria monocytogenes; Listeriosis; Mice; Mice, Inbred C3H; Molecular Sequence Data
PubMed: 12704115
DOI: 10.1128/IAI.71.5.2447-2454.2003 -
Frontiers in Immunology 2020While Baccillus Calmette-Guerin (BCG) is used worldwide, tuberculosis (TB) is still a global concern due to the poor efficacy of BCG. Novel vaccine candidates are...
While Baccillus Calmette-Guerin (BCG) is used worldwide, tuberculosis (TB) is still a global concern due to the poor efficacy of BCG. Novel vaccine candidates are therefore urgently required. In this study, two attenuated recombinant strains, LMΔ and LIΔ were constructed by deletion of and and expression of a fusion protein consisting of T cell epitopes from four () antigens (, and ). The safety and immunogenicity of the two recombinant strains were evaluated in C57BL/6J mice. After intravenous immunization individually, both recombinant strains entered liver and spleen but eventually were eliminated from these organs after several days. Simultaneously, they induced antigen-specific cell-mediated immunity, indicating that the recombinant strains were immunogenic and safe . LMΔ immunization induced stronger cellular immune responses than LIΔ immunization, and when boosted with LIΔ, antigen-specific IFN-γ CD8 T cell responses were notably magnified. Furthermore, we evaluated the protection conferred by the vaccine candidates against mycobacterial infection via challenging the mice with 1 × 10 CFU of BCG. Especially, we tested the feasibility of application of them as heterologous BCG supplement vaccine by immunization of mice with BCG firstly, and boosted with LMΔ and LIΔ sequentially before challenging. Combination immune strategy (LMΔ prime-LIΔ boost) conferred comparable protection efficacy as BCG alone. More importantly, BCG-vaccinated mice acquired stronger resistance to Mycobacterial challenge when boosted with LMΔ and LIΔ sequentially. Our results inferred that heterologous immunization with -based recombinant strains boosted BCG-primed protection against pulmonary mycobacterial infection.
Topics: Animals; Antibodies, Bacterial; Antigens, Bacterial; BCG Vaccine; Cross Reactions; Cytokines; Disease Models, Animal; Female; Genetic Engineering; Immunization, Secondary; Immunoglobulin G; Immunohistochemistry; Immunophenotyping; Listeria; Listeria monocytogenes; Macrophages; Mice; Mycobacterium tuberculosis; T-Lymphocytes; Tuberculosis Vaccines; Tuberculosis, Pulmonary; Virulence
PubMed: 32983151
DOI: 10.3389/fimmu.2020.02036 -
Applied and Environmental Microbiology Sep 1991A sensitive and specific method for detection of Listeria monocytogenes in milk and ground-beef samples is described. It consists of culturing samples in listeria...
A sensitive and specific method for detection of Listeria monocytogenes in milk and ground-beef samples is described. It consists of culturing samples in listeria enrichment broth (LEB) and subculturing them from LEB to listeria plating media, followed by DNA extraction and species-specific detection of the organism by using the polymerase chain reaction (PCR). In developing the L. monocytogenes PCR assay, five oligonucleotide primers complementary to the nucleotide sequence of the listeriolysin O gene were synthesized and used in amplification experiments. PCR products of the predicted size, based on nucleotide sequence information, were generated with DNA from all of 72 L. monocytogenes strains with five different primer pairs. DNA from Listeria ivanovii, Listeria innocua, Listeria seeligeri, Listeria welshimeri, Listeria grayi, and Listeia murrayi strains and a panel of 47 bacterial strains representing 17 genera did not generate PCR products with the primer pairs employed. As little as 1 pg of L. monocytogenes DNA could be detected with the assay. To determine the most sensitive culture protocol to use in conjunction with the PCR assay, milk (10 ml) and ground-beef (25 g) samples were inoculated with L. monocytogenes at concentrations ranging from 0 to 10(5) CFU ml-1 or g-1, as appropriate for the sample. PCR assays on DNA extracted from growth on listeria plating media, inoculated with 24-h LEB samples cultures, were most sensitive, allowing detection of as little as 0.1 CFU of L. monocytogenes ml-1 or g-1 of milk and ground beef, respectively.
Topics: Animals; Bacterial Proteins; Bacterial Toxins; Cattle; Food Microbiology; Heat-Shock Proteins; Hemolysin Proteins; Listeria monocytogenes; Meat; Milk; Polymerase Chain Reaction; Sensitivity and Specificity
PubMed: 1768130
DOI: 10.1128/aem.57.9.2576-2580.1991 -
Prevalence and contamination levels of Listeria monocytogenes in smoked fish and pâté sold in Spain.Journal of Food Protection Dec 2001From March to November 2000, 170 samples of smoked fish and 182 samples of pâté for sale in retail outlets and supermarkets in the nine provinces of Castilla and León...
From March to November 2000, 170 samples of smoked fish and 182 samples of pâté for sale in retail outlets and supermarkets in the nine provinces of Castilla and León (Spain) were analyzed for the prevalence of Listeria monocytogenes and other Listeria spp. L. monocytogenes was isolated from 38 (22.3%) of the 170 samples of smoked fish analyzed. Twenty of these positive samples contained L. monocytogenes at >100 CFU/g. Other Listeria spp., such as Listeria innocua (26 isolates), Listeria grayi (9), Listeria welshimeri (3), Listeria seeligeri (3), and Listeria ivanovii (2), were also detected. L. monocytogenes was isolated from 5.4% of the 182 samples of pâté. Only 1 of the 10 positive samples harbored >100 L. monocytogenes CFU/g. Two other species of Listeria were observed in pâté: L. innocua (12 isolates) and L. grayi (2).
Topics: Animals; Colony Count, Microbial; Fishes; Food Contamination; Food Microbiology; Listeria; Listeria monocytogenes; Prevalence; Smoke; Spain
PubMed: 11770642
DOI: 10.4315/0362-028x-64.12.2075 -
Journal of Food Protection Jun 1996The sensitivity of five strains of Listeria to electron beam irradiation in ground pork as well as the extent of sublethal radiation injury exhibited by each were...
The sensitivity of five strains of Listeria to electron beam irradiation in ground pork as well as the extent of sublethal radiation injury exhibited by each were investigated. Ground pork was inoculated with one of five strains of Listeria and irradiated with from 0 to 1.25 kGy at 0.25 kGy intervals. Listeria innocua NADC 2841 was more radiation-resistant (D = 0.638 kGy) than L. monocytogenes NADC 2045 Scott A (D = 0.447 kGy), L. monocytogenes NADC 2783 (a hamburger isolate) (D = 0.424 kGy), L. monocytogenes ATCC 15313 (D = 0.445 kGy), and L. ivanovii NADC 3518 (D = 0.372 kGy), when recovered on tryptic soy agar supplemented with 0.6% yeast extract. D values for L. innocua , L. ivanovii , and L. monocytogenes ATCC 15313 were lower when cells were recovered on modified Oxford medium. These three strains were susceptible to radiation-induced sublethal injury, with the numbers of injured organisms increasing with irradiation dose. The two pathogenic strains of L. monocytogenes were not injured significantly at the dose levels used. The results show that the dose range currently being considered by the Food and Drug Administration for the irradiation of beef and pork (1.5 to 4.5 kGy) is adequate for the elimination of L. monocytogenes from pork.
PubMed: 31159025
DOI: 10.4315/0362-028X-59.6.596 -
Journal of Food Protection Jul 1994Ewes' milk samples from 287 farm bulk tanks and 17 transport tankers were analyzed for Listeria over a one-year period. Listeria monocytogenes and Listeria innocua were...
Ewes' milk samples from 287 farm bulk tanks and 17 transport tankers were analyzed for Listeria over a one-year period. Listeria monocytogenes and Listeria innocua were detected in 2.19% and 2.00% of 1052 farm samples, and in 18.38% and 11.76% of 136 tanker samples, respectively. Incidence of Listeria grayi , Listeria ivanovii , Listeria seeligeri and Listeria welshimeri was under 0.4% in farm samples and under 1% in tanker samples. Most farms (93.38%) produced milk free from L. monocytogenes throughout the one-year sampling period. No seasonal influence on milk contamination by Listeria was found. However, ewes' milk contamination by L. monocytogenes and other Listeria spp. was significantly higher in farms where cows were also reared than in farms where only ewes were present.
PubMed: 31121711
DOI: 10.4315/0362-028X-57.7.571 -
FEMS Microbiology Reviews Jul 2006Cholesterol-dependent cytolysins (CDCs) are produced by a large number of pathogenic Gram-positive bacteria. Most of these single-chain proteins are secreted in the... (Review)
Review
Cholesterol-dependent cytolysins (CDCs) are produced by a large number of pathogenic Gram-positive bacteria. Most of these single-chain proteins are secreted in the extracellular medium. Among the species producing CDCs, only two species belonging to the genus Listeria (Listeria monocytogenes and Listeria ivanovii) are able to multiply intracellularly and release their toxins in the phagosomal compartment of the infected host cell. This review provides an updated overview on the importance of listeriolysin O (LLO) in the pathogenicity of L. monocytogenes, focusing mainly on two aspects: (1) the structure-function relationship of LLO and (2) its role in intra- and extracellular signalling. We first examine the specific sequence determinants, or protein domains, that make this cytolysin so well adapted to the intracellular lifestyle of L. monocytogenes. The roles that LLO has in cellular signalling events in the context of relations to pathogenesis are also discussed.
Topics: Amino Acid Sequence; Animals; Bacterial Toxins; Heat-Shock Proteins; Hemolysin Proteins; Humans; Immune Tolerance; Listeria monocytogenes; Mice; NF-kappa B; Phagosomes; Signal Transduction; Structure-Activity Relationship
PubMed: 16774585
DOI: 10.1111/j.1574-6976.2006.00021.x -
Infection and Immunity Mar 1996Monoclonal antibodies were generated against a 30-kDa protein fraction derived from culture supernatants of a Listeria monocytogenes strain complemented with additional...
Monoclonal antibodies were generated against a 30-kDa protein fraction derived from culture supernatants of a Listeria monocytogenes strain complemented with additional copies of the prfA regulator gene. Several of the antibodies reacted specifically with a hitherto unidentified, secreted 30-kDa polypeptide. By immunoblot analysis, the expression of this 30kDa polypeptide was found to be dependent on the presence of the PrfA regulator protein. Microsequencing of peptides derived from the partially purified 30-kDa protein revealed homologies to the InlA and InlB polypeptides of L. monocytogenes, which are required for the internalization of the bacteria into nonphagocytic cell lines. This prompted us to term the 30-kDa polypeptide internalin-related protein (Irp). Irp-specific monoclonal antibodies cross-reacted with a 24-kDa polypeptide present in culture supernatants of Listeria ivanovii, indicating the existence of an Irp-related protein in this pathogenic Listeria species.
Topics: Amino Acid Sequence; Antibodies, Monoclonal; Bacterial Proteins; Listeria; Listeria monocytogenes; Membrane Proteins; Molecular Sequence Data; Molecular Weight
PubMed: 8641748
DOI: 10.1128/iai.64.3.1002-1006.1996 -
Microbiology Spectrum Feb 2022Bacteria-derived natural antimicrobial compounds such as bacteriocins, reruterin, and organic acids have recently received substantial attention as food preservatives or...
Bacteria-derived natural antimicrobial compounds such as bacteriocins, reruterin, and organic acids have recently received substantial attention as food preservatives or therapeutic alternatives in human or animal sectors. This study aimed to evaluate the antimicrobial activity of different bacteria-derived antimicrobials, alone or in combination, against a large panel of Gram-negative and Gram-positive bacteria. Bacteriocins, including microcin J25, pediocin PA-1, nisin Z, and reuterin, were investigated alone or in combination with lactic acid and citric acid, using a checkerboard assay. Concentrations were selected based on predetermined MICs against Salmonella enterica subsp. serovar Newport ATCC 6962 and Listeria ivanovii HPB28 as Gram-negative and Gram-positive indicator strains, respectively. The results demonstrated that the combination of microcin J25 + citric acid + lactic acid; microcin J25 + reuterin + citric acid; and microcin J25 + reuterin + lactic acid tested against Newport ATCC 6962 showed synergistic effects (FIC index = 0.5). Moreover, a combination of pediocin PA-1 + citric acid + lactic acid; and reuterin + citric acid + lactic acid against HPB28 showed a partially synergistic interactions (FIC index = 0.75). Nisin Z exerted a partially synergistic effect in combination with acids (FIC index = 0.625 -0.75), whereas when it was combined with reuterin or pediocin PA-1, it showed additive effects (FIC index = 1) against HPB28. The inhibitory activity of synergetic consortia were tested against a large panel of Gram-positive and Gram-negative bacteria. According to our results, combining different antimicrobials with different mechanisms of action led to higher potency and a broad spectrum of inhibition, including multidrug-resistance pathogens. Reuterin and bacteriocins, including microcin J25, pediocin PA-1, nisin were produced and purified with >90% purity. Using the broth-based checkerboard assay the interaction between these compounds (synergetic, additive, or antagonistic) was assessed. By combining different natural antimicrobials with different modes of action and structure (reuteirn, microcin J25, pediocin PA-1, and organic acids), we successfully developed five different synergetic consortia with improved antimicrobial activity and a broad spectrum of inhibition. These consortia were shown to be effective against a large panel of pathogenic and spoilage microorganisms as well as clinically important multidrug-resistance bacteria. Moreover, because the lower concentrations of bacteriocins and reuterin are used in the synergetic consortia, there is a limited risk of toxicity and resistance development for these compounds.
Topics: Anti-Bacterial Agents; Bacterial Infections; Bacteriocins; Drug Resistance, Bacterial; Drug Synergism; Gram-Negative Bacteria; Gram-Positive Bacteria; Humans; Microbial Sensitivity Tests
PubMed: 35170996
DOI: 10.1128/spectrum.00406-21 -
Infection 1988Thiol-activated hemolysins (listeriolysins) from Listeria monocytogenes (Sv4b) and Listeria ivanovii were purified to homogeneity. The N-terminal amino acid sequences of...
Thiol-activated hemolysins (listeriolysins) from Listeria monocytogenes (Sv4b) and Listeria ivanovii were purified to homogeneity. The N-terminal amino acid sequences of the 58 kDa listeriolysin of L. ivanovii and of a 24 kDa protein which may represent the CAMP-factor of L. ivanovii were determined. Antibodies raised against the L. ivanovii listeriolysin and anti-streptolysin O antibodies were used in Western blot analyses to detect listeriolysin(s) in virulent and avirulent Listeria strains. It was found that all virulent strains of L. monocytogenes synthesize and secrete listeriolysin (Mr 58-59 kDa), albeit in significantly variable quantities. No protein cross-reaction with anti-listeriolysin antibodies or anti-streptolysin O-antibodies was present in the supernatant of Listeria innocua, Listeria welshimeri, Listeria grayi and Listeria murrayi strains. Furthermore, the avirulent but hemolytic Listeria seeligeri did not cross-react with these antibodies. In a L. monocytogenes (strain EGD) gene bank constructed in Escherichia coli two types of hemolytic clones were identified. The first type carried recombinant plasmids with a common 2.0 kb fragment coding for a 23 kDa protein. This hemolytic activity was not activated by DTT and the 23 kDa protein did not cross react with anti-listeriolysin or anti-streptolysin antibodies. The other type of hemolytic clones was detected by using anti-streptolysin O antibodies to screen the gene bank. Some of these clones synthesized a protein of 61 kDa which cross reacted with anti-streptolysin O (or anti-listeriolysin) antibodies. By transposon Tn916 mutagenesis of L. monocytogenes two types of nonhemolytic mutants were obtained. Type I produced no extracellular protein that cross reacted with anti-listeriolysin (or anti SLO) antibodies.(ABSTRACT TRUNCATED AT 250 WORDS)
Topics: Amino Acid Sequence; Animals; Antibodies; Antistreptolysin; Bacterial Proteins; Bacterial Toxins; Cross Reactions; Genes, Bacterial; Heat-Shock Proteins; Hemolysin Proteins; Listeria; Listeria monocytogenes; Mice; Molecular Sequence Data; Virulence
PubMed: 3138189
DOI: 10.1007/BF01639739