In Glässer's disease outbreaks, Glaesserella (Haemophilus) parasuis has to overcome the non-specific immune system in the lower respiratory tract, the alveolar macrophages. Here we showed that porcine alveolar macrophages (PAMs) were able to recognize and phagocyte G. parasuis with strain-to-strain variability despite the presence of the capsule in virulent (serovar 1, 5, 12) as well in avirulent strains (serovar 6 and 9). The capsule, outer membrane proteins, virulence-associated autotransporters, cytolethal distending toxins and many other proteins have been identified as virulence factors of this bacterium. Therefore, we immunized pigs with the crude capsular extract (cCE) from the virulent G. parasuis CAPM 6475 strain (serovar 5) and evaluated the role of the anti-cCE/post-vaccinal IgG in the immune response of PAMs to in vitro infection with various G. parasuis strains. We demonstrated the specific binding of the antibodies to the cCE by Western-blotting assay and immunoprecipitation as well as the specific binding to the strain CAPM 6475 in transmission electron microscopy. In the cCE, we identified several virulence-associated proteins that were immunoreactive with IgG isolated from sera of immunized pigs. Opsonization of G. parasuis strains by post-vaccinal IgG led to enhanced phagocytosis of G. parasuis by PAMs at the first two hours of infection. Moreover, opsonization increased the oxidative burst and expression/production of both pro- and anti-inflammatory cytokines. The neutralizing effects of these antibodies on the antioxidant mechanisms of G. parasuis may lead to attenuation of its virulence and pathogenicity in vivo. Together with opsonization of bacteria by these antibodies, the host may eliminate G. parasuis in the infection site more efficiently. Based on these results, the crude capsular extract is a vaccine candidate with immunogenic properties.

• MeSH
• alveolární makrofágy imunologie   metabolismus   mikrobiologie   MeSH
• antigeny bakteriální imunologie   MeSH
• bakteriální pouzdra imunologie   MeSH
• fagocytóza MeSH
• Haemophilus parasuis imunologie   patogenita   MeSH
• hemofilové infekce imunologie   metabolismus   mikrobiologie   MeSH
• kinetika MeSH
• kultivované buňky MeSH
• neutralizující protilátky imunologie   metabolismus   MeSH
• protilátky bakteriální imunologie   metabolismus   MeSH
• reaktivní formy kyslíku metabolismus   MeSH
• séroskupina MeSH
• specificita protilátek MeSH
• Sus scrofa MeSH
• virulence MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Problems with serological cross-reactivity have led to development of a number of PCRs (individual and multiplex) for molecular typing of Actinobacillus pleuropneumoniae, the causative agent of porcine pleuropneumonia. Most of these assays were developed for detection of specific amplicons within capsule biosynthetic genes before the availability of complete sequences for the different serovars. Here we describe comparative analysis of the complete capsular loci for all 18 serovars of A. pleuropneumoniae, and development of two multiplex PCRs for comprehensive capsule typing of this important pig pathogen.

• MeSH
• Actinobacillus pleuropneumoniae klasifikace   genetika   patogenita   MeSH
• bakteriální polysacharidy genetika   MeSH
• bakteriální pouzdra chemie   klasifikace   genetika   MeSH
• infekce bakteriemi rodu Actinobacillus diagnóza   mikrobiologie   MeSH
• multiplexová polymerázová řetězová reakce metody   MeSH
• nemoci prasat diagnóza   mikrobiologie   MeSH
• prasata MeSH
• sekvenční analýza * MeSH
• séroskupina MeSH
• sérotypizace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

The aim of this study was to investigate isolates of Actinobacillus pleuropneumoniae previously designated serologically either as non-typable (NT) or as 'K2:07', which did not produce serovar-specific amplicons in PCR assays. We used whole genome sequencing to identify the capsule (CPS) loci of six previously designated biovar 1 NT and two biovar 1 'K2:O7' isolates of A. pleuropneumoniae from Denmark, as well as a recent biovar 2 NT isolate from Canada. All of the NT isolates have the same six-gene type I CPS locus, sharing common cpsABC genes with serovars 2, 3, 6, 7, 8, 9, 11 and 13. The two 'K2:O7' isolates contain a unique three-gene type II CPS locus, having a cpsA gene similar to that of serovars 1, 4, 12, 14 and 15. The previously NT isolates share the same O-antigen genes, found between erpA and rpsU, as serovars 3, 6, 8, and 15. Whereas the 'K2:O7' isolates, have the same O-antigen genes as serovar 7, which likely contributed to their previous mis-identification. All of the NT and 'K2:O7' isolates have only the genes required for production of ApxII (apxIICA structural genes, and apxIBD export genes). Rabbit polyclonal antisera raised against representative isolates with these new CPS loci demonstrated distinct reactivity compared to the 16 known serovars. The serological and genomic results indicate that the isolates constitute new serovars 17 (previously NT) and 18 (previously 'K2:O7'). Primers designed for amplification of specific serovar 17 and 18 sequences for molecular diagnostics will facilitate epidemiological tracking of these two new serovars of A. pleuropneumoniae.

• MeSH
• Actinobacillus pleuropneumoniae klasifikace   genetika   imunologie   izolace a purifikace   MeSH
• bakteriální pouzdra genetika   MeSH
• DNA bakterií genetika   MeSH
• DNA primery genetika   MeSH
• genotyp * MeSH
• infekce bakteriemi rodu Actinobacillus epidemiologie   veterinární   MeSH
• nemoci prasat epidemiologie   mikrobiologie   MeSH
• polymerázová řetězová reakce metody   MeSH
• prasata MeSH
• sekvenování celého genomu MeSH
• séroskupina * MeSH
• sérotypizace MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• Geografické názvy
• Dánsko epidemiologie   MeSH
• Kanada epidemiologie   MeSH

Humoral deficiencies represent a broad group of disorders. The aim of the study was to compare the levels of antibodies against pneumococcal capsular polysaccharides (anti-PCP) and natural anti-galactosyl (anti-Gal) antibodies in (1) patients with chronic lymphocytic leukaemia (CLL), (2) patients with common variable immunodeficiency (CVID), and (3) a healthy population and to explore their diagnostic and prognostic potential. Serum immunoglobulin levels and levels of anti-Gal IgG, IgA, and IgM and anti-PCP IgG and IgG2 were determined in 59 CLL patients, 30 CVID patients, and 67 healthy controls. Levels of IgG, IgA, IgM, anti-Gal IgA, anti-Gal IgM, and anti-PCP IgA were lower in CLL and CVID patients than in healthy controls (p value for all parameters < 0.0001). Decrease in the levels of IgA, IgM, anti-Gal IgA, and anti-PCP IgA was less pronounced in the CLL group than in the CVID group. IgA decline, anti-Gal IgA, anti-PCP IgA, and anti-PCP IgG2 were negatively correlated with CLL stage. We devise the evaluation of anti-Gal antibodies to be a routine test in humoral immunodeficiency diagnostics, even in cases of immunoglobulin substitution therapy. Significant reductions, mainly in anti-Gal IgA, IgM, and anti-PCP IgA levels, may have prognostic importance in CLL patients.

• MeSH
• autoprotilátky krev   MeSH
• bakteriální pouzdra imunologie   MeSH
• běžná variabilní imunodeficience diagnóza   imunologie   MeSH
• biologické markery krev   MeSH
• dospělí MeSH
• galaktosylceramidy imunologie   MeSH
• humorální imunita MeSH
• imunoglobulin A krev   MeSH
• imunoglobulin G krev   MeSH
• imunoglobulin M krev   MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• pneumokokové infekce diagnóza   imunologie   MeSH
• prognóza MeSH
• protilátky bakteriální krev   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• Streptococcus pneumoniae imunologie   MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

Actinobacillus pleuropneumoniae causes pleuropneumonia, an economically significant lung disease of pigs. Recently, isolates of A. pleuropneumoniae that were serologically distinct from the previously characterized 15 serovars were described, and a proposal was put forward that they comprised a new serovar, serovar 16. Here we used whole-genome sequencing of the proposed serovar 16 reference strain A-85/14 to confirm the presence of a unique capsular polysaccharide biosynthetic locus. For molecular diagnostics, primers were designed from the capsule locus of strain A-85/14, and a PCR was formulated that differentiated serovar 16 isolates from all 15 known serovars and other common respiratory pathogenic/commensal bacteria of pigs. Analysis of the capsule locus of strain A-85/14 combined with the previous serological data show the existence of a sixteenth serovar-designated serovar 16-of A. pleuropneumoniae.

• MeSH
• Actinobacillus pleuropneumoniae klasifikace   genetika   MeSH
• bakteriální pouzdra genetika   MeSH
• diagnostické techniky molekulární metody   MeSH
• DNA bakterií chemie   genetika   MeSH
• DNA primery genetika   MeSH
• genetické lokusy * MeSH
• genom bakteriální MeSH
• infekce bakteriemi rodu Actinobacillus diagnóza   veterinární   MeSH
• nemoci prasat diagnóza   MeSH
• pleuropneumonie mikrobiologie   veterinární   MeSH
• polymerázová řetězová reakce metody   MeSH
• prasata MeSH
• sekvenční analýza DNA MeSH
• séroskupina * MeSH
• zvířata MeSH
• Check Tag
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

Mycobacteria produce a capsule layer, which consists of glycan-like polysaccharides and a number of specific proteins. In this study, we show that, in slow-growing mycobacteria, the type VII secretion system ESX-5 plays a major role in the integrity and stability of the capsule. We have identified PPE10 as the ESX-5 substrate responsible for this effect. Mutants in esx-5 and ppe10 both have impaired capsule integrity as well as reduced surface hydrophobicity. Electron microscopy, immunoblot and flow cytometry analyses demonstrated reduced amounts of surface localized proteins and glycolipids, and morphological differences in the capsular layer. Since capsular proteins secreted by the ESX-1 system are important virulence factors, we tested the effect of the mutations that cause capsular defects on virulence mechanisms. Both esx-5 and ppe10 mutants of Mycobacterium marinum were shown to be impaired in ESX-1-dependent hemolysis. In agreement with this, the ppe10 and esx5 mutants showed reduced recruitment of ubiquitin in early macrophage infection and intermediate attenuation in zebrafish embryos. These results provide a pivotal role for the ESX-5 secretion system and its substrate PPE10, in the capsular integrity of pathogenic mycobacteria. These findings open up new roads for research on the mycobacterial capsule and its role in virulence and immune modulation.

• MeSH
• atypické mykobakteriální infekce metabolismus   MeSH
• bakteriální pouzdra metabolismus   MeSH
• buněčné linie MeSH
• chromatografie na tenké vrstvě MeSH
• dánio pruhované MeSH
• elektronová mikroskopie MeSH
• faktory virulence metabolismus   MeSH
• imunoblotting MeSH
• lidé MeSH
• modely nemocí na zvířatech MeSH
• Mycobacterium marinum metabolismus   patogenita   MeSH
• průtoková cytometrie MeSH
• sekreční systém typu VII metabolismus   MeSH
• virulence fyziologie   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

We used SDS-polyacrylamide gel electrophoresis to investigate the outer membrane proteins (OMPs) band composition of 19 Escherichia coli K1 strains that have spontaneously lost the ability to form K1 polysaccharide capsule (E. coli K1-) and demonstrated different degrees of susceptibility to the bactericidal action of normal human serum. Presented results showed that there were differences between E. coli K1- strains in OMPs expressing capacity. The analysis performed on OMPs has not revealed a direct association between the different OMPs band composition and the susceptibility of these strains to the serum.

• MeSH
• bakteriální pouzdra genetika   MeSH
• baktericidní aktivita krve * MeSH
• elektroforéza v polyakrylamidovém gelu MeSH
• Escherichia coli chemie   izolace a purifikace   fyziologie   MeSH
• infekce močového ústrojí mikrobiologie   MeSH
• infekce vyvolané Escherichia coli mikrobiologie   MeSH
• lidé MeSH
• mikrobiální viabilita MeSH
• proteiny vnější bakteriální membrány analýza   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

New vaccines targeting meningococci expressing serogroup B polysaccharide have been developed, with some being licensed in Europe. Coverage depends on the distribution of disease-associated genotypes, which may vary by age. It is well established that a small number of hyperinvasive lineages account for most disease, and these lineages are associated with particular antigens, including vaccine candidates. A collection of 4,048 representative meningococcal disease isolates from 18 European countries, collected over a 3-year period, were characterized by multilocus sequence typing (MLST). Age data were available for 3,147 isolates. The proportions of hyperinvasive lineages, identified as particular clonal complexes (ccs) by MLST, differed among age groups. Subjects <1 year of age experienced lower risk of sequence type 11 (ST-11) cc, ST-32 cc, and ST-269 cc disease and higher risk of disease due to unassigned STs, 1- to 4-year-olds experienced lower risk of ST-11 cc and ST-32 cc disease, 5- to 14-year-olds were less likely to experience ST-11 cc and ST-269 cc disease, and ≥25-year-olds were more likely to experience disease due to less common ccs and unassigned STs. Younger and older subjects were vulnerable to a more diverse set of genotypes, indicating the more clonal nature of genotypes affecting adolescents and young adults. Knowledge of temporal and spatial diversity and the dynamics of meningococcal populations is essential for disease control by vaccines, as coverage is lineage specific. The nonrandom age distribution of hyperinvasive lineages has consequences for the design and implementation of vaccines, as different variants, or perhaps targets, may be required for different age groups.

• MeSH
• antigeny bakteriální imunologie   MeSH
• bakteriální pouzdra imunologie   MeSH
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• meningokoková meningitida imunologie   mikrobiologie   prevence a kontrola   MeSH
• meningokokové vakcíny imunologie   MeSH
• mladiství MeSH
• mladý dospělý MeSH
• multilokusová sekvenční typizace MeSH
• Neisseria meningitidis séroskupiny B imunologie   MeSH
• Neisseria meningitidis imunologie   izolace a purifikace   MeSH
• předškolní dítě MeSH
• sekvence nukleotidů MeSH
• sekvenční analýza DNA MeSH
• věkové rozložení MeSH
• Check Tag
• dítě MeSH
• dospělí MeSH
• kojenec MeSH
• lidé MeSH
• mladiství MeSH
• mladý dospělý MeSH
• předškolní dítě MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

• Klíčová slova
• Prevenar 13, Pneumo 23,
• MeSH
• bakteriální pouzdra MeSH
• bakteriální vakcíny * terapeutické užití   MeSH
• chronická nemoc * MeSH
• incidence MeSH
• kouření * MeSH
• lidé MeSH
• pneumokokové infekce * diagnóza   epidemiologie   farmakoterapie   imunologie   mortalita   prevence a kontrola   MeSH
• pneumokokové vakcíny * terapeutické užití   MeSH
• rizikové faktory MeSH
• Streptococcus pneumoniae * MeSH
• vakcinace * ekonomika   metody   mortalita   normy   využití   MeSH
• vakcíny konjugované terapeutické užití   MeSH
• věkové faktory * MeSH
• zdravotní stav * MeSH
• Check Tag
• lidé MeSH

The aim of this investigation was to determine the persistence of biofilm-associated antibiotic resistance developed by methicillin-sensitive Staphylococcus aureus (MSSA), of different capsular types, during biofilm formation. Because of superiority of the tissue culture plate (TCP) over the Congo Red Agar (CRA) method for measuring biofilm formation, it was used to determine the persistence of the antibiotic resistance developed by the isolates in biofilms. The antibiotic resistance was found to persist for 3-4 wk post-propagation as planktonic subcultures. Interestingly, some strains even developed resistance to vancomycin and/or teicoplanin. However, no association of either biofilm formation or persistent antibiotic resistance with the major capsular phenotype was observed. These observations highlight the potential significance of (a) determining the antibiograms of S. aureus subcultured from biofilms developed in vitro using the TCP method as well as from planktonic cultures for formulation of an optimal therapeutic strategy, and (b) continuing to identify predominant non-capsular antigens contributing to biofilm formation, regardless of the capsular phenotype for the development of an effective potentially broad-spectrum vaccine for prevention of bovine mastitis caused by S. aureus.

• MeSH
• antibakteriální látky farmakologie   MeSH
• bakteriální léková rezistence * MeSH
• bakteriální pouzdra genetika   MeSH
• bakteriální proteiny MeSH
• biofilmy účinky léků   růst a vývoj   MeSH
• genotyp MeSH
• mastitida skotu mikrobiologie   MeSH
• mikrobiální testy citlivosti MeSH
• polymerázová řetězová reakce MeSH
• skot MeSH
• Staphylococcus aureus účinky léků   izolace a purifikace   fyziologie   MeSH
• zvířata MeSH
• Check Tag
• skot MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Geografické názvy
• Austrálie MeSH