title: Unusual, virulence plasmid-dependent growth behavior of Yersinia enterocolitica in three-dimensional collagen gels.
abstract: As a first approach to establishing a three-dimensional culture infection model, we studied the growth behavior of the extracellular pathogen Yersinia enterocolitica in three-dimensional collagen gels (3D-CoG). Surprisingly, we observed that plasmidless Y. enterocolitica was motile in the 3D-CoG in contrast to its growth in traditional motility agar at 37 degrees C. Motility at 37 degrees C was abrogated in the presence of the virulence plasmid pYV or the exclusive expression of the pYV-located Yersinia adhesion gene yadA. YadA-producing yersiniae formed densely packed (dp) microcolonies, whereas pYVDelta yadA-carrying yersiniae formed loosely packed microcolonies at 37 degrees C in 3D-CoG. Furthermore, we demonstrated that the packing density of the microcolonies was dependent on the head domain of YadA. Moreover, dp microcolony formation did not depend on the capacity of YadA to bind to collagen fibers, as demonstrated by the use of yersiniae producing collagen nonbinding YadA. By using a yopE-gfp reporter, we demonstrated Ca(2+)-dependent expression of this pYV-localized virulence gene by yersiniae in 3D-CoG. In conclusion, this study revealed unique plasmid-dependent growth behavior of yersiniae in a three-dimensional matrix environment that resembles the behavior of yersiniae (e.g., formation of microcolonies) in infected mouse tissue. Thus, this 3D-CoG model may be a first step to a more complex level of in vitro infection models that mimic living tissue, enabling us to study the dynamics of pathogen-host cell interactions.
authors.affiliation: Max von Pettenkofer Institute for Hygiene and Medical Microbiology, Ludwig Maximilians University, D-80336 Munich, Germany. freund@mvp.uni-muenchen.de
authors.forename: Sandra
authors.initials: S
authors.lastname: Freund
authors.forename: Beate
authors.initials: B
authors.lastname: Czech
authors.forename: Konrad
authors.initials: K
authors.lastname: Trülzsch
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Ackermann
authors.forename: Jürgen
authors.initials: J
authors.lastname: Heesemann
mesh_terms: D018829:Adhesins, Bacterial / Q000235:genetics / Q000378:metabolism; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics / Q000378:metabolism; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D003094:Collagen / Q000378:metabolism*; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels / Q000378:metabolism*; D046529:Microscopy, Electron, Transmission; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014774:Virulence / Q000235:genetics; D015008:Yersinia enterocolitica / Q000235:genetics* / Q000472:pathogenicity / Q000648:ultrastructure
chemical_list: D018829:Adhesins, Bacterial; D001425:Bacterial Outer Membrane Proteins; D001426:Bacterial Proteins; D005782:Gels; C064832:YadA protein, Yersinia; C063239:yopE protein, Yersinia; D003094:Collagen