title: Rabbit monoclonal antibodies directed at the T3SS effector protein YopM identify human pathogenic Yersinia isolates.
abstract: The Yersinia outer protein M (YopM) is a type 3 secretion system (T3SS)-dependent effector protein of Yersinia enterocolitica, Yersinia pseudotuberculosis and Yersinia pestis. Although YopM is indispensable for full virulence, its molecular functions still remain largely elusive. Recently, we could identify the recombinant YopM (rYopM) protein derived from the Y. enterocolitica strain 8081 (JB580) as a cell-penetrating protein, which down-regulates the expression of various pro-inflammatory cytokines including TNFα. In this study, we have generated rabbit monoclonal anti-YopM antibodies (RabMabs). RabMabs were characterized by SDS-PAGE and Western blotting using various truncated versions of rYopM to identify epitope-containing domains. RabMabs recognizing either the N- or C-terminus of YopM were characterized further and validated using a collection of 61 pathogenic and non-pathogenic Yersinia strains as well as exemplary strains of major intestinal bacterial pathogens such as Salmonella enterica ssp. enterica, Shigella flexneri and intestinal pathogenic Escherichia coli. RabMab 41.3 directed at the N-terminus of YopM of Y. enterocolitica strain 8081 recognized all YopM-expressing pathogenic Yersinia strains analyzed in this study but failed to recognize non-pathogenic isolates. Thus, RabMab 41.3 might be applicable for the detection of pathogenic Yersinia strains.
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany. Electronic address: rueterc@uni-muenster.de.
authors.forename: Christian
authors.initials: C
authors.lastname: Rüter
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Mariana Ruiz
authors.initials: MR
authors.lastname: Silva
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Grabowski
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Marie-Luise
authors.initials: ML
authors.lastname: Lubos
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Julia
authors.initials: J
authors.lastname: Scharnert
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Marija
authors.initials: M
authors.lastname: Poceva
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany.
authors.forename: Dominik
authors.initials: D
authors.lastname: von Tils
authors.affiliation: Division of Enteropathogenic Bacteria and Salmonella, National Reference Center for Salmonella and other Bacterial Enteric Pathogens, Robert-Koch Institute, Germany.
authors.forename: Antje
authors.initials: A
authors.lastname: Flieger
authors.affiliation: Department of Bacteriology, Max von Pettenkofer Institut, LMU München, Germany.
authors.forename: Jürgen
authors.initials: J
authors.lastname: Heesemann
authors.affiliation: Department of Molecular Genetics and Microbiology and Center for Infectious Diseases, Stony Brook University, New York, USA.
authors.forename: James B
authors.initials: JB
authors.lastname: Bliska
authors.affiliation: Institute of Infectiology - Center for Molecular Biology of Inflammation (ZMBE), University of Münster, Germany. Electronic address: infekt@uni-muenster.de.
authors.forename: M Alexander
authors.initials: MA
authors.lastname: Schmidt
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology / Q000302:isolation & purification; D001425:Bacterial Outer Membrane Proteins / Q000276:immunology*; D005260:Female; D011817:Rabbits; D012680:Sensitivity and Specificity; D015008:Yersinia enterocolitica / Q000276:immunology / Q000302:isolation & purification*; D015010:Yersinia pestis / Q000276:immunology / Q000302:isolation & purification*; D015011:Yersinia pseudotuberculosis / Q000276:immunology / Q000302:isolation & purification*
chemical_list: D000911:Antibodies, Monoclonal; D001425:Bacterial Outer Membrane Proteins; C060772:yopM protein, Yersinia
keywords: Identification of pathogenic Yersinia; N- and C-terminus; RabMabs; Rabbit monoclonal antibodies; YopM