The influence of lipopolysaccharide on the transformation efficiency of Klebsiella pneumoniae

1989 ◽  
Vol 35 (7) ◽  
pp. 735-737 ◽  
Author(s):  
Silvia Camprubí ◽  
Miquel Regué ◽  
Juán Tomás
Keyword(s):  
Klebsiella Pneumoniae ◽  
Significant Influence ◽  
Capsular Polysaccharide ◽  
Transformation Efficiency ◽  
Transformation Procedure ◽  
O Antigen

The presence of O antigen of the lipopolysaccharide on Klebsiella pneumoniae is responsible for the low transformation frequencies observed. No significant influence of capsular polysaccharide could be observed. This phenomenon was independent of the transformation procedure and plasmid size.Key words: lipopolysaccharide, capsular polysaccharide, transformation, Klebsiella pneumoniae.

scholarly journals Development of a new trend conjugate vaccine for the prevention of Klebsiella pneumoniae

2012 ◽  
Vol 4 (1) ◽  
pp. 33 ◽  
Author(s):  
Tarek A. Ahmad ◽  
Medhat Haroun ◽  
Ahmed A. Hussein ◽  
El Sayed H. El Ashry ◽  
Laila H. El-Sayed
Keyword(s):  
Klebsiella Pneumoniae ◽  
Respiratory Diseases ◽  
Capsular Polysaccharide ◽  
Outer Membrane Proteins ◽  
Immunological Methods ◽  
Easy Method ◽  
O Antigen ◽  
Bacterial Endotoxins ◽  
Tract Infections ◽  
Limited Spectrum

<em>Klebsiella pneumoniae</em> is a major cause of nosocomial pneumonia, septicemia and urinary tract infections, especially in newborns, blood cancer patients, and other immunocompromised candidates. The control of <em>K. pneumoniae</em> is a complicated issue due to its tight pathogenesis. Immuno-prophylactic preparations, especially those directed toward the <em>bacterium</em> O-antigen, showed to be the most successful way to prevent the infection incidence. However, all previously proposed preparations were either of limited spectrum or non-maternal, and hence not targeting the main <em>Klebsiella</em> patients. Moreover, all preparations were directed only to prevent the respiratory diseases due to that pathogen. This article addresses the development of a method originally used to purify the non-capsular bacterial- endotoxins, as a new and easy method for vaccine production against <em>K. pneumoniae</em>. The application of this method was preceded by a biotechnological control of capsular polysaccharide production in <em>K. pneumoniae</em>. The new produced natural conjugate between the bacterial O-antigen and its outer membrane proteins was evaluated by physicochemical and immunological methods to investigate its purity, integrity, safety and immunogenicity. It showed to be pure, stable, safe for use, and able to elicit a protective immunoglobulin titer against different <em>Klebsiella</em> infections. This immune-response proved to be transferable to the offspring of the vaccinated experimental rabbits via placenta.

scholarly journals Surface Antigen Exposure by Bismuth Dimercaprol Suppression of Klebsiella pneumoniae Capsular Polysaccharide

1999 ◽  
Vol 67 (2) ◽  
pp. 664-669 ◽  
Author(s):  
Philip Domenico ◽  
J. M. Tomas ◽  
S. Merino ◽  
X. Rubires ◽  
Burke A. Cunha
Keyword(s):  
Membrane Proteins ◽  
Klebsiella Pneumoniae ◽  
Outer Membrane ◽  
Capsular Polysaccharide ◽  
Outer Membrane Proteins ◽  
Defined Medium ◽  
Phagocytic Index ◽  
Dependent Manner ◽  
O Antigen ◽  
Bacterial Capsule

ABSTRACT The bacterial capsule is an important virulence determinant in animal and plant disease. Bacterial capsule and slime can be inhibited by bismuth compounds, especially when complexed with lipophilic thiol chelators. Bismuth dimercaprol (BisBAL) at 1 ppm of Bi3+repressed Klebsiella pneumoniae capsule expression in defined medium by nearly 90%, which exposed subsurface structures. The phagocytic index for BisBAL-treated bacteria increased from <10 to 360 bacteria per 100 neutrophils in the presence of complement and anticapsular or anti-O antigen antiserum. BisBAL treatment also enhanced the reactivity of monoclonal antibodies (MAbs) specific for the O1-antigen lipopolysaccharide (LPS) or the LPS core in a dose-dependent manner as indicated by the results of enzyme-linked immunosorbent assays. When anti-O1 MAb was used, the reactivity increased significantly for fully encapsulated O1:K1 or O1:K2 cells but not for O1:K− cells. Deposition of C3b also increased significantly for BisBAL-treated O1:K1 or O1:K2 cells but not for O1:K− cells. Survival of a serum-sensitive strain was <0.1% when nonimmune human serum absorbed with O1:K1 cells was used and 107% when BisBAL-treated cells were used for absorption. Outer membrane proteins were also more accessible on the surface of K. pneumoniae after BisBAL treatment. Thus, at subinhibitory levels, BisBAL inhibited capsule expression, which promoted phagocytosis, enhanced the reactivity of specific antibodies for LPS O antigen, LPS core epitopes, or outer-membrane proteins, and enhanced complement interaction with encapsulated K. pneumoniae. By unmasking bacterial surface structures and enhancing the immune system reactivity to bacteria, bismuth thiols may prove useful as adjuncts for vaccination.

scholarly journals The Impact of Insertion Sequences on O-Serotype Phenotype and Its O-Locus-Based Prediction in Klebsiella pneumoniae O2 and O1

2020 ◽  
Vol 21 (18) ◽  
pp. 6572
Author(s):  
Daria Artyszuk ◽  
Radosław Izdebski ◽  
Anna Maciejewska ◽  
Marta Kaszowska ◽  
Aleksandra Herud ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Capsular Polysaccharide ◽  
Passive Immunization ◽  
In Silico Analysis ◽  
Clinical Isolates ◽  
Insertion Sequences ◽  
O Antigen ◽  
O Antigens ◽  
The Impact ◽  
K Antigen

Klebsiella pneumoniae is a nosocomial pathogen, pointed out by the World Helth Organisation (WHO) as “critical” regarding the highly limited options of treatment. Lipopolysaccharide (LPS, O-antigen) and capsular polysaccharide (K-antigen) are its virulence factors and surface antigens, determining O- and K-serotypes and encoded by O- or K-loci. They are promising targets for antibody-based therapies (vaccines and passive immunization) as an alternative to antibiotics. To make such immunotherapy effective, knowledge about O/K-antigen structures, drift, and distribution among clinical isolates is needed. At present, the structural analysis of O-antigens is efficiently supported by bioinformatics. O- and K-loci-based genotyping by polymerase chain reaction (PCR) or whole genome sequencing WGS has been proposed as a diagnostic tool, including the Kaptive tool available in the public domain. We analyzed discrepancies for O2 serotyping between Kaptive-based predictions (O2 variant 2 serotype) and the actual phenotype (O2 variant 1) for two K. pneumoniae clinical isolates. Identified length discrepancies from the reference O-locus resulted from insertion sequences (ISs) within rfb regions of the O-loci. In silico analysis of 8130 O1 and O2 genomes available in public databases indicated a broader distribution of ISs in rfbs that may influence the actual O-antigen structure. Our results show that current high-throughput genotyping algorithms need to be further refined to consider the effects of ISs on the LPS O-serotype.

scholarly journals The PTS Components in Klebsiella pneumoniae Affect Bacterial Capsular Polysaccharide Production and Macrophage Phagocytosis Resistance

2021 ◽  
Vol 9 (2) ◽  
pp. 335
Author(s):  
Novaria Sari Dewi Panjaitan ◽  
Yu-Tze Horng ◽  
Chih-Ching Chien ◽  
Hung-Chi Yang ◽  
Ren-In You ◽  
...  
Keyword(s):  
Survival Rate ◽  
Klebsiella Pneumoniae ◽  
Laser Scanning ◽  
Bacterial Resistance ◽  
Capsular Polysaccharide ◽  
Laser Scanning Microscopy ◽  
Confocal Laser ◽  
Intracellular Bacteria ◽  
Wild Type ◽  
Macrophage Phagocytosis

Capsular polysaccharide (CPS) is a crucial virulence factor for Klebsiella pneumoniae infection. We demonstrated an association of CPS production with two phosphoenolpyruvate:carbohydrate phosphotransferase systems (PTSs). Deficiency of crr, encoding enzyme IIA of PTS, in K. pneumoniae enhanced the transcriptional activities of galF, wzi and gnd, which are in the cps gene cluster, leading to high CPS production. A crr mutant exhibited a higher survival rate in 1% hydrogen peroxide than the wild-type. The crr mutant showed less sensitivity to engulfment by macrophage (RAW 264.7) than the wild-type by observing the intracellular bacteria using confocal laser scanning microscopy (CLSM) and by calculating the colony-forming units (CFU) of intracellular bacteria. After long-term incubation, the survival rate of the intracellular crr mutant was higher than that of the wild-type. Deficiency of crr enhanced the transcriptional activities of etcABC which encodes another putative enzyme II complex of a PTS. Deletion of etcABC in the crr mutant reduced CPS production and the transcriptional activities of galF compared to those of the crr mutant. These results indicated that one PTS component, Crr, represses CPS production by repressing another PTS component, EtcABC, in K. pneumoniae. In addition, PTS plays a role in bacterial resistance to macrophage phagocytosis.

scholarly journals Phage Resistance Is Associated with Decreased Virulence in KPC-Producing Klebsiella pneumoniae of the Clonal Group 258 Clade II Lineage

2021 ◽  
Vol 9 (4) ◽  
pp. 762
Author(s):  
Lucia Henrici De Angelis ◽  
Noemi Poerio ◽  
Vincenzo Di Pilato ◽  
Federica De Santis ◽  
Alberto Antonelli ◽  
...  
Keyword(s):  
Klebsiella Pneumoniae ◽  
Parental Strain ◽  
Bacterial Infections ◽  
Capsular Polysaccharide ◽  
Galleria Mellonella ◽  
Bacterial Pathogen ◽  
Phage Resistance ◽  
Infection Model ◽  
Lytic Phage ◽  
Susceptible Host

Phage therapy is now reconsidered with interest in the treatment of bacterial infections. A major piece of information for this application is the definition of the molecular targets exploited by phages to infect bacteria. Here, the genetic basis of resistance to the lytic phage φBO1E by its susceptible host Klebsiella pneumoniae KKBO-1 has been investigated. KKBO-1 phage-resistant mutants were obtained by infection at high multiplicity. One mutant, designated BO-FR-1, was selected for subsequent experiments, including virulence assessment in a Galleria mellonella infection model and characterization by whole-genome sequencing. Infection with BO-FR-1 was associated with a significantly lower mortality when compared to that of the parental strain. The BO-FR-1 genome differed from KKBO-1 by a single nonsense mutation into the wbaP gene, which encodes a glycosyltransferase involved in the first step of the biosynthesis of the capsular polysaccharide (CPS). Phage susceptibility was restored when BO-FR-1 was complemented with the constitutive wbaP gene. Our results demonstrated that φBO1E infects KKBO-1 targeting the bacterial CPS. Interestingly, BO-FR-1 was less virulent than the parental strain, suggesting that in the context of the interplay among phage, bacterial pathogen and host, the emergence of phage resistance may be beneficial for the host.

scholarly journals Amino Acid Substitutions of MagA in Klebsiella pneumoniae Affect the Biosynthesis of the Capsular Polysaccharide

PLoS ONE ◽  
2012 ◽  
Vol 7 (10) ◽  
pp. e46783 ◽  
Author(s):  
Tzu-Lung Lin ◽  
Feng-Ling Yang ◽  
An-Suei Yang ◽  
Hung-Pin Peng ◽  
Tsung-Lin Li ◽  
...  
Keyword(s):  
Amino Acid ◽  
Klebsiella Pneumoniae ◽  
Capsular Polysaccharide ◽  
Amino Acid Substitutions

Surface exposure of the O-antigen in Klebsiella pneumoniae O1:K1 serotype strains

1988 ◽  
Vol 5 (2) ◽  
pp. 141-147 ◽  
Author(s):  
Juan M. Tomas ◽  
Silvia Camprubi ◽  
Paul Williams
Keyword(s):  
Klebsiella Pneumoniae ◽  
O Antigen
Sign in / Sign up

Export Citation Format

Share Document