skip to main content
DOE PAGES title logo U.S. Department of Energy
Office of Scientific and Technical Information

Title: A porcine ligated loop model reveals new insight into the host immune response against Campylobacter jejuni

Abstract

The symptoms of infectious diarrheal disease are mediated by a combination of a pathogen’s 39 virulence factors and the host immune system. Campylobacter jejuni is the leading bacterial 40 cause of diarrhea worldwide due to its near-ubiquitous zoonotic association with poultry. One of 41 the outstanding questions is to what extent the bacteria are responsible for the diarrheal 42 symptoms via intestinal cell necrosis versus immune cell initiated tissue damage. To determine 43 the stepwise process of inflammation that leads to diarrhea, we used a piglet ligated intestinal 44 loop model to study the intestinal response to C. jejuni. Pigs were chosen due to the anatomical 45 similarity between the porcine and the human intestine. We found that the abundance of 46 neutrophil related proteins increased in the intestinal lumen during C. jejuni infection, including 47 proteins related to neutrophil migration (neutrophil elastase and MMP9), actin reorganization 48 (Arp2/3), and antimicrobial proteins (lipocalin-2, myeloperoxidase, S100A8, and S100A9). The 49 appearance of neutrophil proteins also corresponded with increases of the inflammatory 50 cytokines IL-8 and TNF-a. Compared to infection with the C. jejuni wild-type strain, infection with 51 the noninvasive C. jejuni ΔciaD mutant resulted in a blunted inflammatory response, withmore » less 52 inflammatory cytokines and neutrophil markers. Overall, these findings indicate that intestinal 53 inflammation is driven by C. jejuni virulence and that neutrophils are the predominant cell type 54 responding to C. jejuni infection. We propose that this model can be used as a platform to study 55 the early immune events during infection with intestinal pathogens.« less

Authors:
ORCiD logo [1]; ORCiD logo [2];  [1];  [1];  [3];  [4];  [5];  [1];  [1];  [1];  [5]; ORCiD logo [3]; ORCiD logo [2]; ORCiD logo [2]; ORCiD logo [1]
  1. Washington State Univ., Pullman, WA (United States)
  2. Pacific Northwest National Lab. (PNNL), Richland, WA (United States)
  3. US Dept. of Agriculture (USDA), Albany, CA (United States). Agricultural Research Service
  4. US Dept. of Agriculture (USDA), Pullman, WA (United States). Agricultural Research Service; Washington State Univ., Pullman, WA (United States)
  5. US Dept. of Agriculture (USDA), Ames, IA (United States). Agricultural Research Service
Publication Date:
Research Org.:
Pacific Northwest National Lab. (PNNL), Richland, WA (United States). Environmental Molecular Sciences Lab. (EMSL)
Sponsoring Org.:
National Institutes of Health (NIH); USDOE Office of Science (SC), Biological and Environmental Research (BER); USDA; Agricultural Research Service (ARS)
OSTI Identifier:
1671036
Report Number(s):
PNNL-SA-152772
Journal ID: ISSN 1949-0976
Grant/Contract Number:  
AC05-76RL01830; R01AI125356; R21AI124144; GM094623; 5T32AI007025; R01AI083387
Resource Type:
Accepted Manuscript
Journal Name:
Gut Microbes
Additional Journal Information:
Journal Volume: 12; Journal Issue: 1; Journal ID: ISSN 1949-0976
Country of Publication:
United States
Language:
English
Subject:
disease model; innate immunity; proteomics; intestinal disease; pathogenesis

Citation Formats

Negretti, Nicholas M., Ye, Yinyin, Malavasi, Lais M., Pokharel, Swechha M., Huynh, Steven, Noh, Susan, Klima, Cassidy, Gourley, Christopher R., Ragle, Claude A., Bose, Santanu, Looft, Torey, Parker, Craig T., Clair, Geremy C. D., Adkins, Joshua N., and Konkel, Michael E. A porcine ligated loop model reveals new insight into the host immune response against Campylobacter jejuni. United States: N. p., 2020. Web. doi:10.1080/19490976.2020.1814121.
Negretti, Nicholas M., Ye, Yinyin, Malavasi, Lais M., Pokharel, Swechha M., Huynh, Steven, Noh, Susan, Klima, Cassidy, Gourley, Christopher R., Ragle, Claude A., Bose, Santanu, Looft, Torey, Parker, Craig T., Clair, Geremy C. D., Adkins, Joshua N., & Konkel, Michael E. A porcine ligated loop model reveals new insight into the host immune response against Campylobacter jejuni. United States. doi:10.1080/19490976.2020.1814121.
Negretti, Nicholas M., Ye, Yinyin, Malavasi, Lais M., Pokharel, Swechha M., Huynh, Steven, Noh, Susan, Klima, Cassidy, Gourley, Christopher R., Ragle, Claude A., Bose, Santanu, Looft, Torey, Parker, Craig T., Clair, Geremy C. D., Adkins, Joshua N., and Konkel, Michael E. Fri . "A porcine ligated loop model reveals new insight into the host immune response against Campylobacter jejuni". United States. doi:10.1080/19490976.2020.1814121. https://www.osti.gov/servlets/purl/1671036.
@article{osti_1671036,
title = {A porcine ligated loop model reveals new insight into the host immune response against Campylobacter jejuni},
author = {Negretti, Nicholas M. and Ye, Yinyin and Malavasi, Lais M. and Pokharel, Swechha M. and Huynh, Steven and Noh, Susan and Klima, Cassidy and Gourley, Christopher R. and Ragle, Claude A. and Bose, Santanu and Looft, Torey and Parker, Craig T. and Clair, Geremy C. D. and Adkins, Joshua N. and Konkel, Michael E.},
abstractNote = {The symptoms of infectious diarrheal disease are mediated by a combination of a pathogen’s 39 virulence factors and the host immune system. Campylobacter jejuni is the leading bacterial 40 cause of diarrhea worldwide due to its near-ubiquitous zoonotic association with poultry. One of 41 the outstanding questions is to what extent the bacteria are responsible for the diarrheal 42 symptoms via intestinal cell necrosis versus immune cell initiated tissue damage. To determine 43 the stepwise process of inflammation that leads to diarrhea, we used a piglet ligated intestinal 44 loop model to study the intestinal response to C. jejuni. Pigs were chosen due to the anatomical 45 similarity between the porcine and the human intestine. We found that the abundance of 46 neutrophil related proteins increased in the intestinal lumen during C. jejuni infection, including 47 proteins related to neutrophil migration (neutrophil elastase and MMP9), actin reorganization 48 (Arp2/3), and antimicrobial proteins (lipocalin-2, myeloperoxidase, S100A8, and S100A9). The 49 appearance of neutrophil proteins also corresponded with increases of the inflammatory 50 cytokines IL-8 and TNF-a. Compared to infection with the C. jejuni wild-type strain, infection with 51 the noninvasive C. jejuni ΔciaD mutant resulted in a blunted inflammatory response, with less 52 inflammatory cytokines and neutrophil markers. Overall, these findings indicate that intestinal 53 inflammation is driven by C. jejuni virulence and that neutrophils are the predominant cell type 54 responding to C. jejuni infection. We propose that this model can be used as a platform to study 55 the early immune events during infection with intestinal pathogens.},
doi = {10.1080/19490976.2020.1814121},
journal = {Gut Microbes},
number = 1,
volume = 12,
place = {United States},
year = {2020},
month = {9}
}

Journal Article:
Free Publicly Available Full Text
Publisher's Version of Record

Save / Share:

Works referenced in this record:

Blockade of Antimicrobial Proteins S100A8 and S100A9 Inhibits Phagocyte Migration to the Alveoli in Streptococcal Pneumonia
journal, February 2008


Innate immunity in the small intestine
journal, January 2011

  • Santaolalla, Rebeca; Fukata, Masayuki; Abreu, Maria T.
  • Current Opinion in Gastroenterology, Vol. 27, Issue 2
  • DOI: 10.1097/MOG.0b013e3283438dea

The role of neutrophils in the event of intestinal inflammation
journal, December 2009


In-feed bacitracin methylene disalicylate modulates the turkey microbiota and metabolome in a dose-dependent manner
journal, June 2019


Campylobacter jejuni Infection in Broiler Chickens
journal, March 2006

  • Dhillon, A. Singh; Shivaprasad, H. L.; Schaberg, D.
  • Avian Diseases, Vol. 50, Issue 1
  • DOI: 10.1637/7411-071405R.1

Lipooligosaccharide locus class of Campylobacter jejuni: sialylation is not needed for invasive infection
journal, June 2014

  • Ellström, P.; Feodoroff, B.; Hänninen, M. -L.
  • Clinical Microbiology and Infection, Vol. 20, Issue 6
  • DOI: 10.1111/1469-0691.12382

Human IgM paraproteins demonstrate shared reactivity between Campylobacter jejuni lipopolysaccharides and human peripheral nerve disialylated gangliosides
journal, December 1997


Structural Characterization of Campylobacter jejuni Lipooligosaccharide Outer Cores Associated with Guillain-Barré and Miller Fisher Syndromes
journal, January 2007

  • Godschalk, Peggy C. R.; Kuijf, Mark L.; Li, Jianjun
  • Infection and Immunity, Vol. 75, Issue 3
  • DOI: 10.1128/IAI.00872-06

Pseudaminic Acid on Campylobacter jejuni Flagella Modulates Dendritic Cell IL-10 Expression via Siglec-10 Receptor: A Novel Flagellin-Host Interaction
journal, May 2014

  • Stephenson, Holly N.; Mills, Dominic C.; Jones, Hannah
  • The Journal of Infectious Diseases, Vol. 210, Issue 9
  • DOI: 10.1093/infdis/jiu287

A re-examination of the role of Campylobacter fetus subspecies coli in enteric disease of the pig
journal, August 1981


A direct-sensing galactose chemoreceptor recently evolved in invasive strains of Campylobacter jejuni
journal, October 2016

  • Day, Christopher J.; King, Rebecca M.; Shewell, Lucy K.
  • Nature Communications, Vol. 7, Issue 1
  • DOI: 10.1038/ncomms13206

Epithelial cells secrete the chemokine interleukin-8 in response to bacterial entry.
journal, January 1993


The Reactome Pathway Knowledgebase
journal, November 2017

  • Fabregat, Antonio; Jupe, Steven; Matthews, Lisa
  • Nucleic Acids Research, Vol. 46, Issue D1
  • DOI: 10.1093/nar/gkx1132

The Intestinal Microbiota Influences Campylobacter jejuni Colonization and Extraintestinal Dissemination in Mice
journal, May 2015

  • O'Loughlin, Jason L.; Samuelson, Derrick R.; Braundmeier-Fleming, Andrea G.
  • Applied and Environmental Microbiology, Vol. 81, Issue 14
  • DOI: 10.1128/AEM.00281-15

MPLEx: a Robust and Universal Protocol for Single-Sample Integrative Proteomic, Metabolomic, and Lipidomic Analyses
journal, May 2016


Role of the small Rho GTPases Rac1 and Cdc42 in host cell invasion of Campylobacter jejuni
journal, October 2007


Naive Bayesian Classifier for Rapid Assignment of rRNA Sequences into the New Bacterial Taxonomy
journal, June 2007

  • Wang, Q.; Garrity, G. M.; Tiedje, J. M.
  • Applied and Environmental Microbiology, Vol. 73, Issue 16
  • DOI: 10.1128/AEM.00062-07

MRP-8 and MRP-14, two abundant Ca 2+ -binding proteins of neutrophils and monocytes
journal, February 1993

  • Hessian, Paul A.; Edgeworth, Jonathan; Hogg, Nancy
  • Journal of Leukocyte Biology, Vol. 53, Issue 2
  • DOI: 10.1002/jlb.53.2.197

Campylobacter jejuni FlpA Binds Fibronectin and Is Required for Maximal Host Cell Adherence
journal, October 2009

  • Konkel, M. E.; Larson, C. L.; Flanagan, R. C.
  • Journal of Bacteriology, Vol. 192, Issue 1
  • DOI: 10.1128/JB.00969-09

Chemically Etched Open Tubular and Monolithic Emitters for Nanoelectrospray Ionization Mass Spectrometry
journal, November 2006

  • Kelly, Ryan T.; Page, Jason S.; Luo, Quanzhou
  • Analytical Chemistry, Vol. 78, Issue 22, p. 7796-7801
  • DOI: 10.1021/ac061133r

Mucosal Lipocalin 2 Has Pro-Inflammatory and Iron-Sequestering Effects in Response to Bacterial Enterobactin
journal, October 2009


Campylobacter jejuni Colonization of Mice with Limited Enteric Flora
journal, September 2006

  • Chang, Christopher; Miller, Jeff F.
  • Infection and Immunity, Vol. 74, Issue 9
  • DOI: 10.1128/IAI.01094-05

Protective and pathogenic functions of macrophage subsets
journal, October 2011

  • Murray, Peter J.; Wynn, Thomas A.
  • Nature Reviews Immunology, Vol. 11, Issue 11
  • DOI: 10.1038/nri3073

Characterization and remediation of sample index swaps by non-redundant dual indexing on massively parallel sequencing platforms
journal, May 2018


Characterization of the oxidative stress stimulon and PerR regulon of Campylobacter jejuni
journal, January 2009


Cutting Edge: Bacterial Flagellin Activates Basolaterally Expressed TLR5 to Induce Epithelial Proinflammatory Gene Expression
journal, August 2001


The food-borne pathogen Campylobacter jejuni responds to the bile salt deoxycholate with countermeasures to reactive oxygen species
journal, November 2017

  • Negretti, Nicholas M.; Gourley, Christopher R.; Clair, Geremy
  • Scientific Reports, Vol. 7, Issue 1
  • DOI: 10.1038/s41598-017-15379-5

The focal complex of epithelial cells provides a signalling platform for interleukin-8 induction in response to bacterial pathogens: Pathogen-mediated IL-8 induction
journal, June 2014

  • Eucker, Tyson P.; Samuelson, Derrick R.; Hunzicker-Dunn, Mary
  • Cellular Microbiology, Vol. 16, Issue 9
  • DOI: 10.1111/cmi.12305

Identification of a fibronectin-binding domain within the Campylobacter jejuni CadF protein: C. jejuni fibronectin-binding protein
journal, July 2005


Campylobacter colitis: histological immunohistochemical and ultrastructural findings.
journal, September 1985

  • van Spreeuwel, J. P.; Duursma, G. C.; Meijer, C. J.
  • Gut, Vol. 26, Issue 9
  • DOI: 10.1136/gut.26.9.945

Avian Intestinal Mucus Modulates Campylobacter jejuni Gene Expression in a Host-Specific Manner
journal, January 2019


The SILVA ribosomal RNA gene database project: improved data processing and web-based tools
journal, November 2012

  • Quast, Christian; Pruesse, Elmar; Yilmaz, Pelin
  • Nucleic Acids Research, Vol. 41, Issue D1
  • DOI: 10.1093/nar/gks1219

The Polysaccharide Capsule of Campylobacter jejuni Modulates the Host Immune Response
journal, December 2012

  • Maue, Alexander C.; Mohawk, Krystle L.; Giles, David K.
  • Infection and Immunity, Vol. 81, Issue 3
  • DOI: 10.1128/IAI.01008-12

Campylobacter jejuni Infections: Update on Emerging Issues and Trends
journal, April 2001

  • Acheson, D.; Allos, B. M.
  • Clinical Infectious Diseases, Vol. 32, Issue 8
  • DOI: 10.1086/319760

Biochemical characterization of the neutrophil-specific antigen NB1
journal, February 1990


g:Profiler: a web server for functional enrichment analysis and conversions of gene lists (2019 update)
journal, May 2019

  • Raudvere, Uku; Kolberg, Liis; Kuzmin, Ivan
  • Nucleic Acids Research, Vol. 47, Issue W1
  • DOI: 10.1093/nar/gkz369

Experimental Campylobacter jejuni Infection in Humans
journal, March 1988

  • Black, R. E.; Levine, M. M.; Clements, M. L.
  • Journal of Infectious Diseases, Vol. 157, Issue 3
  • DOI: 10.1093/infdis/157.3.472

Interferon-gamma directly affects barrier function of cultured intestinal epithelial monolayers.
journal, February 1989

  • Madara, J. L.; Stafford, J.
  • Journal of Clinical Investigation, Vol. 83, Issue 2
  • DOI: 10.1172/JCI113938

Campylobacter jejuni-Stimulated Secretion of Interleukin-8 by INT407 Cells
journal, January 1999


Central role of IL-17/Th17 immune responses and the gut microbiota in the pathogenesis of intestinal fibrosis
journal, January 2014


Translocation of Campylobacter jejuni across Human Polarized Epithelial Cell Monolayer Cultures
journal, August 1992

  • Konkel, M. E.; Mead, D. J.; Hayes, S. F.
  • Journal of Infectious Diseases, Vol. 166, Issue 2
  • DOI: 10.1093/infdis/166.2.308

The Complete Campylobacter jejuni Transcriptome during Colonization of a Natural Host Determined by RNAseq
journal, August 2013


Campylobacter Enteritis: Clinical and Epidemiologic Features
journal, August 1979


Peripheral CD4+ T Cell Cytokine Responses Following Human Challenge and Re-Challenge with Campylobacter jejuni
journal, November 2014


Fibronectin-Facilitated Invasion of T84 Eukaryotic Cells by Campylobacter jejuni Occurs Preferentially at the Basolateral Cell Surface
journal, December 2002


Evaluation of Agar Plates for Direct Enumeration of Campylobacter spp. from Poultry Carcass Rinses
journal, June 2005

  • Oyarzabal, Omar A.; Macklin, Kenneth S.; Barbaree, James M.
  • Applied and Environmental Microbiology, Vol. 71, Issue 6
  • DOI: 10.1128/AEM.71.6.3351-3354.2005

The Campylobacter jejuni CiaD effector protein activates MAP kinase signaling pathways and is required for the development of disease
journal, January 2013

  • Samuelson, Derrick R.; Eucker, Tyson P.; Bell, Julia A.
  • Cell Communication and Signaling, Vol. 11, Issue 1
  • DOI: 10.1186/1478-811X-11-79

Newborn piglet model for campylobacteriosis.
journal, January 1993


Integrin activation by the lipid molecule 25-hydroxycholesterol induces a proinflammatory response
journal, April 2019


Molecular Dissection of the Campylobacter jejuni CadF and FlpA Virulence Proteins in Binding to Host Cell Fibronectin
journal, March 2020


Identification of a Campylobacter jejuni -secreted protein required for maximal invasion of host cells
journal, August 2009


DADA2: High-resolution sample inference from Illumina amplicon data
journal, May 2016

  • Callahan, Benjamin J.; McMurdie, Paul J.; Rosen, Michael J.
  • Nature Methods, Vol. 13, Issue 7
  • DOI: 10.1038/nmeth.3869

The Perseus computational platform for comprehensive analysis of (prote)omics data
journal, June 2016

  • Tyanova, Stefka; Temu, Tikira; Sinitcyn, Pavel
  • Nature Methods, Vol. 13, Issue 9
  • DOI: 10.1038/nmeth.3901

CheY‐mediated modulation of Campylobacter jejuni virulence
journal, March 1997


Characterization of a porcine intestinal epithelial cell line for in vitro studies of microbial pathogenesis in swine
journal, October 2005

  • Schierack, Peter; Nordhoff, Marcel; Pollmann, Marion
  • Histochemistry and Cell Biology, Vol. 125, Issue 3
  • DOI: 10.1007/s00418-005-0067-z

Increased expression of surface activation markers on neutrophils following migration into the nasal lumen
journal, August 2003


How Intestinal Bacteria Cause Disease
journal, March 1999

  • Guerrant, Richard L.; Steiner, Ted S.; Lima, Aldo A. M.
  • The Journal of Infectious Diseases, Vol. 179, Issue s2
  • DOI: 10.1086/513845

Chemotactic behavior of Campylobacter jejuni.
journal, January 1988


Human Intestinal Barrier Function in Health and Disease
journal, January 2016

  • König, Julia; Wells, Jerry; Cani, Patrice D.
  • Clinical and Translational Gastroenterology, Vol. 7, Issue 10
  • DOI: 10.1038/ctg.2016.54

Quantifying and Understanding Well-to-Well Contamination in Microbiome Research
journal, June 2019


Acute diarrhoea: Campylobacter colitis and the role of rectal biopsy.
journal, October 1979

  • Price, A. B.; Jewkes, J.; Sanderson, P. J.
  • Journal of Clinical Pathology, Vol. 32, Issue 10
  • DOI: 10.1136/jcp.32.10.990

The Campylobacter jejuni CiaC virulence protein is secreted from the flagellum and delivered to the cytosol of host cells
journal, January 2012

  • Neal-McKinney, Jason M.; Konkel, Michael E.
  • Frontiers in Cellular and Infection Microbiology, Vol. 2
  • DOI: 10.3389/fcimb.2012.00031

Myeloperoxidase: a front-line defender against phagocytosed microorganisms
journal, October 2012

  • Klebanoff, S. J.; Kettle, A. J.; Rosen, H.
  • Journal of Leukocyte Biology, Vol. 93, Issue 2
  • DOI: 10.1189/jlb.0712349

Human S100A9 potentiates IL-8 production in response to GM-CSF or fMLP via activation of a different set of transcription factors in neutrophils
journal, May 2014


Serine phosphorylation of cortactin is required for maximal host cell invasion by Campylobacter jejuni
journal, January 2013

  • Samuelson, Derrick R.; Konkel, Michael E.
  • Cell Communication and Signaling, Vol. 11, Issue 1
  • DOI: 10.1186/1478-811X-11-82

Proinflammatory Activities of S100: Proteins S100A8, S100A9, and S100A8/A9 Induce Neutrophil Chemotaxis and Adhesion
journal, March 2003


The MaxQuant computational platform for mass spectrometry-based shotgun proteomics
journal, October 2016


Absorption of bile salts from the small intestine in vivo
journal, January 1962


The Host Antimicrobial Protein Calgranulin C Participates in the Control of Campylobacter jejuni Growth via Zinc Sequestration
journal, April 2018

  • Shank, Janette M.; Kelley, Brittni R.; Jackson, Joseph W.
  • Infection and Immunity, Vol. 86, Issue 6
  • DOI: 10.1128/IAI.00234-18

Reconstitution of a Functional Toll-like Receptor 5 Binding Site in Campylobacter jejuni Flagellin
journal, February 2010

  • de Zoete, Marcel R.; Keestra, A. Marijke; Wagenaar, Jaap A.
  • Journal of Biological Chemistry, Vol. 285, Issue 16
  • DOI: 10.1074/jbc.M109.070227

Attachment, Invasion, and Translocation of Campylobacter jejuni in Pig Small-Intestinal Epithelial Cells
journal, May 2010

  • Pogačar, Maja Šikić; Klančnik, Anja; Možina, Sonja Smole
  • Foodborne Pathogens and Disease, Vol. 7, Issue 5
  • DOI: 10.1089/fpd.2009.0301