title: Anti-LcrV antibody inhibits delivery of Yops by Yersinia pestis KIM5 by directly promoting phagocytosis.
abstract: LcrV of Yersinia pestis is a major protective antigen proposed for inclusion in subunit plague vaccines. One way that anti-LcrV antibody is thought to protect is by inhibiting the delivery of toxins called Yops to host cells. The present study characterizes the relation between this inhibition and the phagocytosis of the bacteria. J774A.1 cells were infected with Y. pestis KIM5 in the presence of a protective polyclonal anti-LcrV antibody or a nonprotective polyclonal anti-YopM antibody, and delivery of YopH and YopE into the cytoplasm was assayed by immunoblotting. The ability to inhibit the delivery of these Yops depended upon having antibody bound to the cell surface; blocking conditions that prevented the binding of antibody to Fc receptors prevented the inhibition of Yop delivery. Anti-LcrV antibody also promoted phagocytosis of the yersiniae, whereas F(ab')(2) fragments did not. Further, anti-LcrV antibody could not inhibit the delivery of Yops into cells that were unable to phagocytose due to the presence of cytochalasin D. However, Yops were produced only by extracellular yersiniae. We hypothesize that anti-LcrV antibody does not directly inhibit Yop delivery but instead causes phagocytosis, with consequent inhibition of Yop protein production in the intracellular yersiniae. The prophagocytic effect of anti-LcrV antibody extended to mouse polymorphonuclear neutrophils (PMNs) in vitro, and PMNs were shown to be critical for protection: when PMNs in mice were ablated, the mice lost all ability to be protected by anti-LcrV antibody.
authors.affiliation: Department of Microbiology, Immunology, and Molecular Genetics, University of Kentucky, Lexington, KY 40536-0298, USA.
authors.forename: Clarissa
authors.initials: C
authors.lastname: Cowan
authors.forename: Alexander V
authors.initials: AV
authors.lastname: Philipovskiy
authors.forename: Christine R
authors.initials: CR
authors.lastname: Wulff-Strobel
authors.forename: Zhan
authors.initials: Z
authors.lastname: Ye
authors.forename: Susan C
authors.initials: SC
authors.lastname: Straley
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000494:pharmacology*; D000942:Antigens, Bacterial / Q000276:immunology*; D001425:Bacterial Outer Membrane Proteins / Q000037:antagonists & inhibitors* / Q000276:immunology / Q000378:metabolism; D006367:HeLa Cells; D006801:Humans; D007140:Immunoglobulin Fab Fragments / Q000276:immunology / Q000494:pharmacology; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins / Q000037:antagonists & inhibitors* / Q000378:metabolism; D051379:Mice; D009504:Neutrophils / Q000276:immunology / Q000382:microbiology; D010587:Phagocytosis / Q000276:immunology*; D010930:Plague / Q000276:immunology* / Q000378:metabolism; D052899:Pore Forming Cytotoxic Proteins; D015010:Yersinia pestis / Q000276:immunology* / Q000378:metabolism
chemical_list: D000907:Antibodies, Bacterial; D000942:Antigens, Bacterial; D001425:Bacterial Outer Membrane Proteins; D007140:Immunoglobulin Fab Fragments; C043908:LcrV protein, Yersinia; D049990:Membrane Transport Modulators; D026901:Membrane Transport Proteins; D052899:Pore Forming Cytotoxic Proteins; C094424:YscC protein, Yersinia pestis; C060772:yopM protein, Yersinia