title: Comparative proteomic analysis reveals activation of mucosal innate immune signaling pathways during cholera.
abstract: Vibrio cholerae O1 is a major cause of acute watery diarrhea in over 50 countries. Evidence suggests that V. cholerae O1 may activate inflammatory pathways, and a recent study of a Bangladeshi population showed that variants in innate immune genes play a role in mediating susceptibility to cholera. We analyzed human proteins present in the small intestine of patients infected with V. cholerae O1 to characterize the host response to this pathogen. We collected duodenal biopsy specimens from patients with acute cholera after stabilization and again 30 days after initial presentation. Peptides extracted from biopsy specimens were sequenced and quantified using label-free mass spectrometry and SEQUEST. Twenty-seven host proteins were differentially abundant between the acute and convalescent stages of infection; the majority of these have known roles in innate defense, cytokine production, and apoptosis. Immunostaining confirmed that two proteins, WARS and S100A8, were more abundant in lamina propria cells during the acute stage of cholera. Analysis of the differentially abundant proteins revealed the activation of key regulators of inflammation by the innate immune system, including Toll-like receptor 4, nuclear factor kappa-light-chain-enhancer of activated B cells, mitogen-activated protein kinases, and caspase-dependent inflammasomes. Interleukin-12β (IL-12β) was a regulator of several proteins that were activated during cholera, and we confirmed that IL-12β was produced by lymphocytes recovered from duodenal biopsy specimens of cholera patients. Our study shows that a broad inflammatory response is generated in the gut early after onset of cholera, which may be critical in the development of long-term mucosal immunity against V. cholerae O1.
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA cnellis@mgh.harvard.edu jbharris@mgh.harvard.edu.
authors.forename: Crystal N
authors.initials: CN
authors.lastname: Ellis
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Regina C
authors.initials: RC
authors.lastname: LaRocque
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Taher
authors.initials: T
authors.lastname: Uddin
authors.affiliation: ThermoFischer Scientific, Biomarker Research in Mass Spectrometry, Cambridge, Massachusetts, USA.
authors.forename: Bryan
authors.initials: B
authors.lastname: Krastins
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA.
authors.forename: Leslie M
authors.initials: LM
authors.lastname: Mayo-Smith
authors.affiliation: ThermoFischer Scientific, Biomarker Research in Mass Spectrometry, Cambridge, Massachusetts, USA.
authors.forename: David
authors.initials: D
authors.lastname: Sarracino
authors.affiliation: Center for Systems Biology, Department of Organismic and Evolutionary Biology, Harvard University, Cambridge, Massachusetts, USA Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
authors.forename: Elinor K
authors.initials: EK
authors.lastname: Karlsson
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Atiqur
authors.initials: A
authors.lastname: Rahman
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Tahmina
authors.initials: T
authors.lastname: Shirin
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Taufiqur R
authors.initials: TR
authors.lastname: Bhuiyan
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Fahima
authors.initials: F
authors.lastname: Chowdhury
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Ashraful Islam
authors.initials: AI
authors.lastname: Khan
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA Department of Immunology and Infectious Diseases, Harvard School of Public Health, Boston, Massachusetts, USA.
authors.forename: Edward T
authors.initials: ET
authors.lastname: Ryan
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA Department of Medicine, Harvard Medical School, Boston, Massachusetts, USA Department of Microbiology and Immunobiology, Harvard Medical School, Boston, Massachusetts, USA.
authors.forename: Stephen B
authors.initials: SB
authors.lastname: Calderwood
authors.affiliation: International Centre for Diarrhoeal Disease Research, Dhaka, Bangladesh.
authors.forename: Firdausi
authors.initials: F
authors.lastname: Qadri
authors.affiliation: Division of Infectious Diseases, Massachusetts General Hospital, Boston, Massachusetts, USA Department of Pediatrics, Harvard Medical School, Boston, Massachusetts, USA cnellis@mgh.harvard.edu jbharris@mgh.harvard.edu.
authors.forename: Jason B
authors.initials: JB
authors.lastname: Harris
mesh_terms: D000208:Acute Disease; D017209:Apoptosis / Q000276:immunology; D001706:Biopsy; D040501:Calgranulin A / Q000235:genetics / Q000276:immunology; D002771:Cholera / Q000235:genetics* / Q000276:immunology / Q000382:microbiology / Q000473:pathology; D003289:Convalescence; D004386:Duodenum / Q000276:immunology* / Q000382:microbiology / Q000473:pathology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D054884:Host-Pathogen Interactions; D006801:Humans; D018928:Immunity, Mucosal; D058847:Inflammasomes / Q000235:genetics / Q000276:immunology; D053762:Interleukin-12 Subunit p40 / Q000235:genetics / Q000276:immunology; D040901:Proteomics; D015398:Signal Transduction / Q000276:immunology*; D051197:Toll-Like Receptor 4 / Q000235:genetics / Q000276:immunology; D014369:Tryptophan-tRNA Ligase / Q000235:genetics / Q000276:immunology; D041241:Vibrio cholerae O1 / Q000254:growth & development / Q000276:immunology / Q000472:pathogenicity*
chemical_list: D040501:Calgranulin A; D058847:Inflammasomes; D053762:Interleukin-12 Subunit p40; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; D014369:Tryptophan-tRNA Ligase; C073175:WARS1 protein, human