Isolation of a Bacteriophage and Its Depolymerase Specific for K1 Capsule of Klebsiella pneumoniae: Implication in Typing and Treatment

Lin, Tzu‐Lung; Hsieh, Pei-Fang; Huang, Yu-Tsung; Lee, Wei-Ching; Tsai, Yi‐Lin; Su, Po-An; Pan, Yi-Jiun; Hsu, Chun-Ru; Wu, Mao S.; Wang, Jann-Tay

doi:10.1093/infdis/jiu332

Cited by 128 publications

(148 citation statements)

References 42 publications

(52 reference statements)

Supporting

Mentioning

139

Contrasting

Order By: Relevance

“…Consequently, several techniques for molecular capsular typing were developed to circumvent these problems (7,9,39,40). In addition, phage-borne capsular polysaccharide depolymerases recently were used in capsular typing of Klebsiella (6,22). The identification of capsule depolymerases in ⌽K64-1 with specificity for K1, K11, K21, K25, K30/K69, K35, K64, KN4, and KN5 could serve as the basis for further rapid and simple approaches for the characterization of these capsular types.…”

Section: Discussionmentioning

confidence: 99%

“…Klebsiella phages have been reported since 1940 and have been used for determination of several K-types of Klebsiella (16)(17)(18)(19)(20)(21). However, little was known about the host specificity determinants of these phages until the recent characterization and identification of bacteriophageborne depolymerases (6,22). A KN2-specific phage, 0507-KN2-1, and its capsule depolymerase were identified and used for capsular typing of the new KN2 capsular type (6).…”

mentioning

confidence: 99%

“…A KN2-specific phage, 0507-KN2-1, and its capsule depolymerase were identified and used for capsular typing of the new KN2 capsular type (6). Another study documented a phage (NTUH-K2044-K1-1) and its capsule depolymerase that exhibited specificity for capsular type K1 and showed therapeutic efficacy in K1 K. pneumoniae-infected mice (22). We recently isolated a multihost Klebsiellainfecting bacteriophage, ⌽K64-1 (8), which appeared to belong to the Myoviridae family of viruses based on the sequence similarity.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Klebsiella Phage ΦK64-1 Encodes Multiple Depolymerases for Multiple Host Capsular Types

Pan

Lin

Chen

et al. 2017

J Virol

Self Cite

112

View full text Add to dashboard Cite

The genome of the multihost bacteriophage ⌽K64-1, capable of infecting Klebsiella capsular types K1, K11, K21, K25, K30, K35, K64, and K69, as well as new capsular types KN4 and KN5, was analyzed and revealed that 11 genes (S1-1, S1-2, S1-3, S2-1, S2-2, S2-3, S2-4, S2-5, S2-6, S2-7, and S2-8) encode proteins with amino acid sequence similarity to tail fibers/spikes or lyases. S2-5 previously was shown to encode a K64 capsule depolymerase (K64dep). Specific capsule-degrading activities of an additional eight putative capsule depolymerases (S2-4 against K1, S1-1 against K11, S1-3 against K21, S2-2 against K25, S2-6 against K30/K69, S2-3 against K35, S1-2 against KN4, and S2-1 against KN5) was demonstrated by expression and purification of the recombinant proteins. Consistent with the capsular type-specific depolymerization activity of these gene products, phage mutants of S1-2, S2-2, S2-3, or S2-6 lost infectivity for KN4, K25, K35, or K30/K69, respectively, indicating that capsule depolymerase is crucial for infecting specific hosts. In conclusion, we identified nine functional capsule depolymerase-encoding genes in a bacteriophage and correlated activities of the gene products to all ten hosts of this phage, providing an example of type-specific host infection mechanisms in a multihost bacteriophage.IMPORTANCE We currently identified eight novel capsule depolymerases in a multihost Klebsiella bacteriophage and correlated the activities of the gene products to all hosts of this phage, providing an example of carriage of multiple depolymerases in a phage with a wide capsular type host spectrum. Moreover, we also established a recombineering system for modification of Klebsiella bacteriophage genomes and demonstrated the importance of capsule depolymerase for infecting specific hosts. Based on the powerful tool for modification of phage genome, further studies can be conducted to improve the understanding of mechanistic details of Klebsiella phage infection. Furthermore, the newly identified capsule depolymerases will be of great value for applications in capsular typing.KEYWORDS Klebsiella, bacteriophage, capsular type, capsule depolymerase, multiple host T he genus Klebsiella, especially the species Klebsiella pneumoniae, is an important human pathogen that causes a wide range of diseases, including both community and hospital-acquired infections. It is associated with septicemia, pneumonia, and urinary tract infections (1, 2) and also is responsible for a globally emerging disease, pyogenic liver abscess complicated with metastatic meningitis and endophthalmitis (3,4).Klebsiella spp. typically display a layer of thick, polysaccharide-based capsule on their surfaces. The expression of diverse capsule structure caused by different sugar compositions and linkages divide them into distinct serotypes. In addition, genetic

show abstract

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Klebsiella Phage ΦK64-1 Encodes Multiple Depolymerases for Multiple Host Capsular Types

Pan

Lin

Chen

et al. 2017

J Virol

Self Cite

112

View full text Add to dashboard Cite

show abstract

“…Since K64 strains were less virulent than strains causing community-acquired pyogenic liver abscess (PLA) (for example, the 50% lethal dose [LD 50 ] of NTUH-K2044 for mice was ϳ1 ϫ 10 3 CFU [20]) and exhibited very high lethal doses (LD 50 of 3 ϫ 10 7 CFU), we assessed the therapeutic effect of capsule depolymerase in mice treated with cyclophosphamide, which reduces the numbers of white blood cells in mice (20). The results showed that 6 of the 8 mice infected with 6 ϫ 10 6 CFU of a CRKP K64 strain (KCR2A) died within 2 days without enzyme treatment.…”

Section: Figmentioning

confidence: 99%

Identification of Capsular Types in Carbapenem-Resistant Klebsiella pneumoniae Strains by wzc Sequencing and Implications for Capsule Depolymerase Treatment

Pan

Lin

et al. 2015

Antimicrob Agents Chemother

Self Cite

100

113

View full text Add to dashboard Cite

dKlebsiella pneumoniae is an important human pathogen associated with a variety of diseases, and the prevalence of multidrugresistant K. pneumoniae (MDRKP) is rapidly increasing. Here we determined the capsular types of 85 carbapenem-resistant K. pneumoniae (CRKP) strains by wzc sequencing and investigated the presence of carbapenemases and integrons among CRKP strains. Ten CRKP strains (12%) were positive for carbapenemase (imipenemase, 6/85 strains; K. pneumoniae carbapenemase, 3/85 strains; Verona integron-encoded metallo-␤-lactamase, 1/85 strains). Capsular type K64 accounted for 32 CRKP strains (38%), followed by K62 (13%), K24 (8%), KN2 (7%), and K28 (6%). Sequence types (STs) were determined by multilocus sequence typing (MLST), and the results indicated that ST11, which accounted for 47% of these CRKP strains (40/85 strains), was the major ST. We further isolated a K64-specific capsule depolymerase (K64dep), which could enhance serum and neutrophil killing in vitro and increase survival rates for K64 K. pneumoniae-inoculated mice. The toxicity study demonstrated that mice treated with K64dep showed normal biochemical parameters and no significant histopathological changes of liver, kidney, and spleen, indicating that enzyme treatment did not cause toxicity in mice. Therefore, the findings of capsular type clustering among CRKP strains and effective treatment with capsule depolymerase for MDRKP infections are important for capsule-based vaccine development and therapy.K lebsiella pneumoniae, a Gram-negative bacillus, causes hospital or community-acquired disease (1-3). Most K. pneumoniae strains harbor chromosome-encoded SHV ␤-lactamase (4). In 1983, the emergence of K. pneumoniae strains producing a mutant of SHV-1 ␤-lactamase that hydrolyzes extended-spectrum cephalosporins was observed in Germany (5); subsequently, K. pneumoniae strains resistant to third-generation cephalosporins were detected in France, and a new ␤-lactamase gene closely related to TEM-1 and TEM-2 was identified (6). In 1989, the CTX-M type was reported as a new extended-spectrum ␤-lactamase (ESBL) family member not belonging to either the TEM type or the SHV type (7), and subsequently it was considered one of the major ESBL types (8). Recent global surveillance data from Europe, North and South America, and Asia revealed that the frequency of ESBL-producing K. pneumoniae was 7.5 to 44% (9). As the prevalence of ESBL-producing isolates increased, carbapenems were used to treat serious infections caused by ESBL-producing K. pneumoniae. However, carbapenem-resistant K. pneumoniae (CRKP) rates have been dramatically increasing worldwide over the past 10 years. In the United States, a significant increase in CRKP prevalence from Ͻ1% in 2000 to 8% in 2007 was reported (10). In addition, data from the National Healthcare Safety Network (NHSN) showed that, in 2006 and 2007, CRKP was reported for up to 10.8% of total isolates associated with certain device-related infections (11). In Italy, CRKP increased rapidly from 1 to 2% in 2006 to 2009...

show abstract

“…The enzyme also prevented dissemination, sterilized the blood, and promoted early recovery in nonhuman primates infected by the intratracheal and intrabronchial routes (20). In addition to our previous work on systemic neonatal E. coli infections (9, 10), capsule depolymerases have been shown to resolve potentially lethal experimental Klebsiella pneumoniae K1 infections in mice (21).…”

Section: Discussionmentioning

confidence: 88%

Parenteral Administration of Capsule Depolymerase EnvD Prevents Lethal Inhalation Anthrax Infection

Negus

Vipond

Hatch

et al. 2015

Antimicrob Agents Chemother

View full text Add to dashboard Cite

Left untreated, inhalation anthrax is usually fatal. Vegetative forms of Bacillus anthracis survive in blood and tissues during infection due to elaboration of a protective poly-␥-D-glutamic acid (PDGA) capsule that permits uncontrolled bacterial growth in vivo, eventually leading to overwhelming bacillosis and death. As a measure to counter threats from multidrug-resistant strains, we are evaluating the prophylactic and therapeutic potential of the PDGA depolymerase EnvD, a stable and potent enzyme which rapidly and selectively removes the capsule from the surface of vegetative cells. Repeated intravenous administration of 10 mg/kg recombinant EnvD (rEnvD) to mice infected with lethal doses of B. anthracis Ames spores by inhalation prevented the emergence of symptoms of anthrax and death; all animals survived the 5-day treatment period, and 70% survived to the end of the 14-day observation period. In contrast to results in sham-treated animals, the lungs and spleen of rEnvD-dosed animals were free of gross pathological changes. We conclude that rEnvD has potential as an agent to prevent the emergence of inhalation anthrax in infected animals and is likely to be effective against drug-resistant forms of the pathogen.

show abstract

Isolation of a Bacteriophage and Its Depolymerase Specific for K1 Capsule of Klebsiella pneumoniae: Implication in Typing and Treatment

Abstract: These results demonstrate this phage and its capsule depolymerase exhibit specificity for capsular type K1 and can be used for the diagnosis and treatment of K1 K. pneumoniae infections.

Cited by 128 publications

References 42 publications

Klebsiella Phage ΦK64-1 Encodes Multiple Depolymerases for Multiple Host Capsular Types

Klebsiella Phage ΦK64-1 Encodes Multiple Depolymerases for Multiple Host Capsular Types

Identification of Capsular Types in Carbapenem-Resistant Klebsiella pneumoniae Strains by wzc Sequencing and Implications for Capsule Depolymerase Treatment

Parenteral Administration of Capsule Depolymerase EnvD Prevents Lethal Inhalation Anthrax Infection

Contact Info

Product

Resources

About