Identification of potential vaccine target antigens by immunoproteomic analysis of a virulent and a non-virulent strain of the fish pathogen Flavobacterium psychrophilum

Sudheesh, Ponnerassery S.; LaFrentz, Benjamin R.; Call, Douglas R.; Siems, William F.; LaPatra, Scott E.; Wiens, Gregory D.; Cain, Kenneth D.

doi:10.3354/dao074037

Cited by 51 publications

(53 citation statements)

References 59 publications

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“…Previous research in our laboratory has identified another protein spot at ~58 kDa as OmpA (P60) (Fig. 4), which was immunogenic and exhibited experimental values that are closer to the theoretical molecular mass and pI of the protein (LaFrentz 2007). Similarly, Dumetz et al (2008) identified multiple protein spots with approximate molecular masses of < 30 kDa as OmpA (P60), and they suggested that these represented degraded OmpA (P60) products from sample preparation.…”

Section: Discussionsupporting

confidence: 60%

“…Research suggests that this protein may be involved in the virulence of bacterial pathogens (Chastanet et al 2004, Yuan et al 2007). The ClpB protein of Francisella tularensis is immunogenic (Havlasova et al 2005), and similarly the ClpB protein of Flavobacterium psychrophilum is immunogenic (LaFrentz 2007).…”

Section: Discussionmentioning

confidence: 99%

“…A previously frozen glycerol stock of a virulent strain of Flavobacterium psychrophilum, CSF-259-93 (Sudheesh et al 2007), was plated for isolation on tryptone yeast extract salts (TYES; 0.4% tryptone, 0.04% yeast extract, 0.05% MgSO 4 , 0.05% CaCl 2 , pH 7.2) agar (Holt et al 1993) and incubated at 15°C for 4 d. Several colonies were transferred to 20 ml TYES broth and incubated at 15°C for 72 h. The 72 h cultures were used in the following growth conditions. Growth in iron-limited conditions: A trial experiment was implemented to determine the optimal concentration of the iron chelator 2, 2-dipyridyl (DPD; Sigma-Aldrich) to obtain inhibited growth of Flavobacterium psychrophilum.…”

Section: Methodsmentioning

confidence: 99%

“…Therefore, recent efforts have been aimed at identifying specific bacterial antigens to target for vaccine development. Such research has resulted in the identification of numerous proteins or specific fractions of F. psychrophilum (Rahman et al 2002, Merle et al 2003, LaFrentz et al 2004, Massias et al 2004, Crump et al 2005, Dumetz et al 2007, 2008, LaFrentz 2007, Sudheesh et al 2007), some of which have been shown to be protective (Rahman et al 2002, LaFrentz et al 2004, Dumetz et al 2006, Crump et al 2007). These studies demonstrate the potential for identifying vaccine candidates; however, the work was performed using in vitro cultured bacteria, and our knowledge of differential gene expression by F. psychrophilum grown in vivo is lacking.…”

Section: Introductionmentioning

confidence: 99%

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Proteomic analysis of Flavobacterium psychrophilum cultured in vivo and in iron-limited media

LaFrentz

LaPatra²,

Call³

et al. 2009

Dis. Aquat. Org.

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Flavobacterium psychrophilum is the etiologic agent of bacterial coldwater disease, but the pathogenic mechanisms of this important fish pathogen are not fully understood. Identifying bacterial genes of F. psychrophilum differentially expressed in vivo may lead to a better understanding of pathogenesis and provide targets for vaccine development. Therefore, the present study used a proteomic approach to identify and quantify proteins of F. psychrophilum following growth in vivo and under iron-limited growth conditions. As determined by 2D polyacrylamide gel electrophoresis (2D-PAGE), numerous proteins exhibited different spot intensities following culture of the bacterium in vivo, and of these, 20 were selected and identified by liquid chromatography-mass spectrometry/ mass spectrometry (LC-MS/MS) analysis and Mascot searches of the F. psychrophilum genome. Eighteen proteins exhibited increased spot intensities in vivo, and these included: several chaperone and stress proteins, gliding motility protein GldN, outer membrane protein OmpH, 2 probable outer membrane proteins (OmpA family), probable aminopeptidase precursor, probable lipoprotein precursor, 3-oxoacyl-[acyl-carrier-protein]-reductase, and several proteins with unknown function. Two proteins exhibited decreased spot intensities in vivo and were identified as ferritin FtnA and outer membrane protein OmpA (P60). Culture of F. psychrophilum in iron-limited media resulted in similar protein spot intensity changes for 6 of the 20 proteins identified following growth in vivo. Results from the present study suggest a role of upregulated proteins in the pathogenesis of F. psychrophilum and these may represent potential vaccine candidate antigens.

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Section: Discussionsupporting

confidence: 60%

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Proteomic analysis of Flavobacterium psychrophilum cultured in vivo and in iron-limited media

LaFrentz

LaPatra²,

Call³

et al. 2009

Dis. Aquat. Org.

Self Cite

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“…ECPs produced by the bacterial cells were prepared by the cellophane overlay method according to Sudheesh et al (2007), with some modifications. The bacterial cells grown on TYES agar were suspended in 0.5% NaCl and spread onto sterile cellophane (Sigma-Aldrich)-overlaid TYES agar plates.…”

Section: Preparation Of Extracellular Products (Ecp)mentioning

confidence: 99%

Phase variation in Flavobacterium psychrophilum: characterization of two distinct colony phenotypes

Högfors‐Rönnholm

Wıklund²

2010

Dis. Aquat. Org.

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Four 'smooth' and 4 'rough' colony phenotypes of the Gram-negative fish pathogen Flavobacterium psychrophilum isolated from rainbow trout Oncorhynchus mykiss were characterized using biochemical, physiological, molecular and virulence tests to better understand the pathogenesis of the bacterium. Biochemically, the 2 cell types did not react significantly differently. Physiologically, the 2 phenotypes had distinct characteristics, and, when grown in broth, the smooth cells were found to be autoagglutinating and able to switch into the non-agglutinating rough phenotype. The rough cells did not switch into the smooth phenotype under any growth conditions tested, indicating that the phase variation from the smooth to rough phenotype is irreversible or that the conditions for the reversible switch are still to be found. Smooth cells were hydrophobic and more adhesive compared to the hydrophilic rough cells, suggesting that the phase variation most probably involves one or several surface structures other than outer membrane proteins and lipopolysaccharides that were found to be similar in both types. Analysis of extracellular products produced by the 2 cell types indicated furthermore that a difference in enzymatic activities could exist. Both cell types were virulent for rainbow trout in an intramuscular challenge; thus, the distinct physiological characteristics of the phenotypes do not seem to be directly associated with virulence, when the body surface of the fish is disregarded. The results suggest that phase variation occurs in F. psychrophilum, but that the importance of the 2 phenotypes for the pathogenesis of the bacterium has still to be investigated.

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Application of Proteomics to Elucidate Bacterium–Host Interactions

Smith

2011

Methods in Animal Proteomics

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Identification of potential vaccine target antigens by immunoproteomic analysis of a virulent and a non-virulent strain of the fish pathogen Flavobacterium psychrophilum

Cited by 51 publications

References 59 publications

Proteomic analysis of Flavobacterium psychrophilum cultured in vivo and in iron-limited media

Proteomic analysis of Flavobacterium psychrophilum cultured in vivo and in iron-limited media

Phase variation in Flavobacterium psychrophilum: characterization of two distinct colony phenotypes

Application of Proteomics to Elucidate Bacterium–Host Interactions

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