Proteomic analysis of Lawsonia intracellularis reveals expression of outer membrane proteins during infection

Watson, E.D.; Alberdi, Pilar; Inglis, Neil F.; Lainson, Alex F.; Porter, Megan E.; Manson, Erin; Imrie, Lisa; McLean, Kevin; Smith, David George Emslie

doi:10.1016/j.vetmic.2014.10.002

Cited by 17 publications

(7 citation statements)

References 35 publications

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“…LatA was shown to specifically react to sera from infected pigs. Another mass spectrometry study utilizing a shotgun proteomics approach identified five outer membrane proteins of L. intracellularis , of which two were antigenic and reacted with infected pig serum ( 41 ). A putative hemolysin of L. intracellularis named LhlyA has been examined at a preliminary level ( 42 ).…”

Section: Introductionmentioning

confidence: 99%

Lawsonia intracellularis: Revisiting the Disease Ecology and Control of This Fastidious Pathogen in Pigs

Karuppannan

Opriessnig

2018

Front. Vet. Sci.

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Lawsonia intracellularis is an anaerobic obligate intracellular bacterium infecting the small intestine and infrequently also the large intestine of pigs and other animals including hamsters and horses. The infection is characterized by proliferation, hemorrhage, necrosis, or any combination commonly referred to as “ileitis,” affecting the health and production efficacy of farmed pigs. Despite decades of research on this pathogen, the pathogenesis and virulence factors of this organism are not clearly known. In pigs, prophylaxis against L. intracellularis infection is achieved by either administration of subtherapeutic levels of in-feed antibiotic growth promoters or vaccination. While the former approach is considered to be effective in L. intracellularis control, potential regulations on subtherapeutic antibiotics in many countries in the near future may necessitate alternative approaches. The potential of manipulating the gut microbiome of pigs with feed ingredients or supplements to control L. intracellularis disease burden is promising based on the current understanding of the porcine gut microbiome in general, as well as preliminary insights into the disease ecology of L. intracellularis infection accrued over the last 30 years.

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Section: Introductionmentioning

confidence: 99%

Lawsonia intracellularis: Revisiting the Disease Ecology and Control of This Fastidious Pathogen in Pigs

Karuppannan

Opriessnig

2018

Front. Vet. Sci.

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“…4E), of which 45 (84.9%) were confidently predicted by bioinformatics (Table 3). Of previous studies that have utilised gel-based proteomics to identify OMPs, Watson et al [55] identified 19 OMPs in Lawsonia intracellularis , Wang et al [50] identified 19 OMPs in E. tarda and Ayalew et al [52] identified 55 OMPs in M. haemolytica . Using 2-D gel-based proteomics, Coquet et al [65] identified 55 OMPs in Y. ruckeri in a comparison of bacteria grown in planktonic culture and as a biofilm.…”

Section: Discussionmentioning

confidence: 99%

“…Subcellular fractionation of the outer membrane followed by gel-free proteomics has been successfully employed for the identification of OMPs in Edwardsiella tarda [49], Coxiella burnetti [51] , P. multocida [34] and Mannheimia haemolytica [52]. Similarly, gel-based proteomic approaches have been extensively used to characterise bacterial whole-cells, secreted proteins, OMVs and OMPs [[53], [54], [55], [56]]. Although few studies have combined both gel-free and gel-based approaches to characterise the outer membrane, coverage has generally been maximised by the use of both complementary techniques [[57], [58], [59], [60], [61], [62], [63]].…”

Section: Introductionmentioning

confidence: 99%

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Ormsby

Grahame

Burchmore

et al. 2019

Journal of Proteomics

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Yersinia ruckeri is the aetiological agent of enteric redmouth (ERM) disease and is responsible for significant economic losses in farmed salmonids. Enteric redmouth disease is associated primarily with rainbow trout ( Oncorhynchus mykiss, Walbaum) but its incidence in Atlantic salmon ( Salmo salar ) is increasing. Outer membrane proteins (OMPs) of Gram-negative bacteria are located at the host-pathogen interface and play important roles in virulence. The outer membrane of Y. ruckeri is poorly characterised and little is known about its composition and the roles of individual OMPs in virulence. Here, we employed a bioinformatic pipeline to first predict the OMP composition of Y. ruckeri . Comparative proteomic approaches were subsequently used to identify those proteins expressed in vitro in eight representative isolates recovered from Atlantic salmon and rainbow trout. One hundred and forty-one OMPs were predicted from four Y. ruckeri genomes and 77 of these were identified in three or more genomes and were considered as “core” proteins. Gel-free and gel-based proteomic approaches together identified 65 OMPs in a single reference isolate and subsequent gel-free analysis identified 64 OMPs in the eight Atlantic salmon and rainbow trout isolates. Together, our gel-free and gel-based proteomic analyses identified 84 unique OMPs in Y. ruckeri . Significance Yersinia ruckeri is an important pathogen of Atlantic salmon and rainbow trout and is of major economic significance to the aquaculture industry worldwide. Disease outbreaks are becoming more problematic in Atlantic salmon and there is an urgent need to investigate in further detail the cell-surface (outer membrane) composition of strains infecting each of these host species. Currently, the outer membrane of Y. ruckeri is poorly characterised and very little is known about the OMP composition of strains infecting each of these salmonid species. This study represents the most comprehensive comparative outer membrane proteomic analysis of Y. ruckeri to date, encompassing isolates of different biotypes, serotypes, OMP-types and hosts of origin and provides insights into the potential roles of these diverse proteins in host-pathogen interactions. The study has identified key OMPs likely to be involved in disease pathogenesis and makes a significant contribution to furthering our understanding of the cell-surface composition of this important fish pathogen that will be relevant to the development of improved vaccines and therapeutics.

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“…Despite the completion of LI genome sequencing, protein immunogens against LI infection have yet to be characterized. Proteomic and comparative transcriptional analyses were recently conducted to identify functions related to the pathogenic mechanisms of the LI proteins [ 7 , 21 ].…”

Section: Discussionmentioning

confidence: 99%

Antigenic and functional profiles of a Lawsonia intracellularis protein that shows a flagellin-like trait and its immuno-stimulatory assessment

Won

Lee

2018

Vet Res

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The obligate intracellular Lawsonia intracellularis (LI), the etiological agent of proliferative enteropathy (PE), is an economically important disease in the swine industry. Due to extreme difficulty of in vitro culture of the pathogen, molecular characterization of protein components of LI that are targets of the immune system, is difficult; thus, the scientific evidence to drive the development of preventive measures is lacking. In this work, we investigated the antigenic and functional characteristics of a putative flagellar-associated protein, LI0570, using in silico computational approaches for epitope prediction and an in vitro protein-based molecular assay. The amino acid sequence of LI0570 exhibited similarities to flagellar-associated proteins in four different bacterial strains. The presence of B cell linear confirmative epitopes of the protein predicted by a bioinformatics tool was validated by western blot analysis using anti-LI mouse hyperimmune serum, which implied that LI0570 induced production of antigen-specific antibodies in vivo. Further, TLR5-stimulating activity and IL-8 cytokine expression produced via downstream signaling were observed in HEK-Blue™-hTLR5 cells stimulated with LI0570. This result indicates that the LI0570 protein can trigger an innate immune response followed by a T-cell-related adaptive immune response in an infected host. Collectively, the data presented here support that the LI0570 protein which shows the antigenic potential could be a useful component of a recombinant vaccine against PE, providing progress toward an effective prevention strategy.

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Proteomic analysis of Lawsonia intracellularis reveals expression of outer membrane proteins during infection

Cited by 17 publications

References 35 publications

Lawsonia intracellularis: Revisiting the Disease Ecology and Control of This Fastidious Pathogen in Pigs

Lawsonia intracellularis: Revisiting the Disease Ecology and Control of This Fastidious Pathogen in Pigs

Comparative bioinformatic and proteomic approaches to evaluate the outer membrane proteome of the fish pathogen Yersinia ruckeri

Antigenic and functional profiles of a Lawsonia intracellularis protein that shows a flagellin-like trait and its immuno-stimulatory assessment

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