Proteomic analysis of porcine mesenteric lymph-nodes after Salmonella typhimurium infection

Martins, Rodrigo Prado; Collado-Romero, Melania; Martínez-Gomáriz, Montserrat; Carvajal, Ana; Gil, Concha; Lucena, Carlos; Moreno, A.; Garrido, Juan J.

doi:10.1016/j.jprot.2012.03.045

Cited by 21 publications

(23 citation statements)

References 42 publications

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“…The UniProt Knowledgebase (KB) version used in this study contained only 1,406 manually curated, nonredundant (SwissProt) entries for Sus scrofa proteins, compared with 20,248 entries in the human and 65,102 in the mammalian databases (54). To overcome the paucity of fully annotated entries, MS data from porcine samples are often searched against either human plus pig or mammalian protein databases (6,35,56,70). An additional UniProtKB database, TrEMBL, contains protein sequences associated with computationally generated annotation but is not reviewed nor curated for redundancy.…”

Section: Discussionmentioning

confidence: 99%

Protein composition of bronchoalveolar lavage fluid and airway surface liquid from newborn pigs

Bartlett

Albertolle

Wohlford‐Lenane

et al. 2013

American Journal of Physiology-Lung Cellular and Molecular Phys

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The airway mucosa and the alveolar surface form dynamic interfaces between the lung and the external environment. The epithelial cells lining these barriers elaborate a thin liquid layer containing secreted peptides and proteins that contribute to host defense and other functions. The goal of this study was to develop and apply methods to define the proteome of porcine lung lining liquid, in part, by leveraging the wealth of information in the Sus scrofa database of Ensembl gene, transcript, and protein model predictions. We developed an optimized workflow for detection of secreted proteins in porcine bronchoalveolar lavage (BAL) fluid and in methacholine-induced tracheal secretions [airway surface liquid (ASL)]. We detected 674 and 3,858 unique porcine-specific proteins in BAL and ASL, respectively. This proteome was composed of proteins representing a diverse range of molecular classes and biological processes, including host defense, molecular transport, cell communication, cytoskeletal, and metabolic functions. Specifically, we detected a significant number of secreted proteins with known or predicted roles in innate and adaptive immunity, microbial killing, or other aspects of host defense. In greatly expanding the known proteome of the lung lining fluid in the pig, this study provides a valuable resource for future studies using this important animal model of pulmonary physiology and disease.

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Section: Discussionmentioning

confidence: 99%

Protein composition of bronchoalveolar lavage fluid and airway surface liquid from newborn pigs

Bartlett

Albertolle

Wohlford‐Lenane

et al. 2013

American Journal of Physiology-Lung Cellular and Molecular Phys

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“…Subsequently, protein from individual replicates belonging to the same group was pooled (30 ug total), electrophoretically fractionated in 12% (w/v) SDS-PAGE gels and transferred onto a PVDF membrane (Millipore, Bedford, MA, USA). Western blot assays were carried out as described by Martins et al [10] employing the following primary antibodies: 4B7/8 for swine histocompatibility class I antigen (SLAI) detection [19], 1 F12 for swine histocompatibility class II antigen (SLAII) detection [19], anti-CTLA4 (Epitomics, Burlingame, CA, USA) and anti-Clathrin light chain (ab24579, Abcam, Cambridge, UK). To confirm equal sample loading, membranes were reblotted with anti-GAPDH monoclonal antibody (GenScript, Picastaway, NJ, USA) and no statistical differences for GAPDH abundance were observed between groups in all assays.…”

Section: Methodsmentioning

confidence: 99%

“…To confirm equal sample loading, membranes were reblotted with anti-GAPDH monoclonal antibody (GenScript, Picastaway, NJ, USA) and no statistical differences for GAPDH abundance were observed between groups in all assays. Membranes were scanned in an FLA-5100 imager (Fujifilm, Tokyo, Japan) and signal intensity was determined using Multigauge software (Fujifilm, Tokyo, Japan) as previously described [10]. …”

Section: Methodsmentioning

confidence: 99%

“…For immunohistochemistry assays, a standard avidin-biotin peroxidase method was performed as described elsewhere [20] employing 1 F12 monoclonal antibody and a biotinylated anti-mouse Ig (Dako, Barcelona, Spain) as a secondary antibody. Immunofluorescence using confocal microscopy was performed employing the anti-SLAII 1 F12 monoclonal antibody, a rabbit polyclonal antibody against the Salmonella somatic (O4, 5, 12) antigen [10] and a rabbit polyclonal antibody anti- Salmonella Typhimurium flagellin [21]. Fluorescein isothiocyanate (FITC)-conjugated goat anti-rabbit IgG (Sigma-Aldrich, St. Louis, MO, USA) and Alexa Fluor 594 anti-mouse IgG (Life Technologies, Carlsbad, CA, USA) were used as secondary antibodies.…”

Section: Methodsmentioning

confidence: 99%

“…Thus, whole-genome expression analysis has been used to explore gene expression changes during infection of pigs by Salmonella , contributing to identify molecules and pathways associated with the host response to infection [7,8]. More recently, proteomic techniques have also been employed as a step towards a detailed understanding of the disease mechanisms [9,10]. However, despite this, there is a need to deepen understanding of the biological processes that control host-pathogen interaction and Salmonella persistence in porcine lymphatic tissue, which could provide new targets for treatment and control of salmonellosis in this species.…”

Section: Introductionmentioning

confidence: 99%

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Pyroptosis and adaptive immunity mechanisms are promptly engendered in mesenteric lymph-nodes during pig infections with Salmonella enterica serovar Typhimurium

Martins

Aguilar

Graham

et al. 2013

Vet Res

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In this study, we explored the transcriptional response and the morphological changes occurring in porcine mesenteric lymph-nodes (MLN) along a time course of 1, 2 and 6 days post infection (dpi) with Salmonella Typhimurium. Additionally, we analysed the expression of some Salmonella effectors in tissue to complete our view of the processes triggered in these organs upon infection. The results indicate that besides dampening apoptosis, swine take advantage of the flagellin and prgJ expression by Salmonella Typhimuriun to induce pyroptosis in MLN, preventing bacterial dissemination. Furthermore, cross-presentation of Salmonella antigens was inferred as a mechanism that results in a rapid clearance of pathogen by cytotoxic T cells. In summary, although the Salmonella Typhimurium strain employed in this study was able to express some of its major virulence effectors in porcine MLN, a combination of early innate and adaptive immunity mechanisms might overcome virulence strategies employed by the pathogen, enabling the host to protect itself against bacterial spread beyond gut-associated lymph-nodes. Interestingly, we deduced that clathrin-mediated endocytosis could contribute to mechanisms of pathogen virulence and/or host defence in MLN of Salmonella infected swine. Taken together, our results are useful for a better understanding of the critical protective mechanisms against Salmonella that occur in porcine MLN to prevent the spread of infection beyond the intestine.

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Proteomics in farm animals models of human diseases

Ceciliani

Restelli

Lecchi

2014

Proteomics Clinical Apps

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The need to provide in vivo complex environments to understand human diseases strongly relies on the use of animal models, which traditionally include small rodents and rabbits. It is becoming increasingly evident that the few species utilised to date cannot be regarded as universal. There is a great need for new animal species that are naturally endowed with specific features relevant to human diseases. Farm animals, including pigs, cows, sheep and horses, represent a valid alternative to commonly utilised rodent models. There is an ample scope for the application of proteomic techniques in farm animals, and the establishment of several proteomic maps of plasma and tissue has clearly demonstrated that farm animals provide a disease environment that closely resembles that of human diseases. The present review offers a snapshot of how proteomic techniques have been applied to farm animals to improve their use as biomedical models. Focus will be on specific topics of biomedical research in which farm animal models have been characterised through the application of proteomic techniques.

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Proteomic analysis of porcine mesenteric lymph-nodes after Salmonella typhimurium infection

Cited by 21 publications

References 42 publications

Protein composition of bronchoalveolar lavage fluid and airway surface liquid from newborn pigs

Protein composition of bronchoalveolar lavage fluid and airway surface liquid from newborn pigs

Pyroptosis and adaptive immunity mechanisms are promptly engendered in mesenteric lymph-nodes during pig infections with Salmonella enterica serovar Typhimurium

Proteomics in farm animals models of human diseases

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