title: Equid herpesvirus type 1 activates platelets.
abstract: Equid herpesvirus type 1 (EHV-1) causes outbreaks of abortion and neurological disease in horses. One of the main causes of these clinical syndromes is thrombosis in placental and spinal cord vessels, however the mechanism for thrombus formation is unknown. Platelets form part of the thrombus and amplify and propagate thrombin generation. Here, we tested the hypothesis that EHV-1 activates platelets. We found that two EHV-1 strains, RacL11 and Ab4 at 0.5 or higher plaque forming unit/cell, activate platelets within 10 minutes, causing α-granule secretion (surface P-selectin expression) and platelet microvesiculation (increased small events double positive for CD41 and Annexin V). Microvesiculation was more pronounced with the RacL11 strain. Virus-induced P-selectin expression required plasma and 1.0 mM exogenous calcium. P-selectin expression was abolished and microvesiculation was significantly reduced in factor VII- or X-deficient human plasma. Both P-selectin expression and microvesiculation were re-established in factor VII-deficient human plasma with added purified human factor VIIa (1 nM). A glycoprotein C-deficient mutant of the Ab4 strain activated platelets as effectively as non-mutated Ab4. P-selectin expression was abolished and microvesiculation was significantly reduced by preincubation of virus with a goat polyclonal anti-rabbit tissue factor antibody. Infectious virus could be retrieved from washed EHV-1-exposed platelets, suggesting a direct platelet-virus interaction. Our results indicate that EHV-1 activates equine platelets and that α-granule secretion is a consequence of virus-associated tissue factor triggering factor X activation and thrombin generation. Microvesiculation was only partly tissue factor and thrombin-dependent, suggesting the virus causes microvesiculation through other mechanisms, potentially through direct binding. These findings suggest that EHV-1-induced platelet activation could contribute to the thrombosis that occurs in clinically infected horses and provides a new mechanism by which viruses activate hemostasis.
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: Tracy
authors.initials: T
authors.lastname: Stokol
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: Wee Ming
authors.initials: WM
authors.lastname: Yeo
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: Deborah
authors.initials: D
authors.lastname: Burnett
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: Nicole
authors.initials: N
authors.lastname: DeAngelis
authors.affiliation: Institut für Virologie, Freie Universität Berlin, Berlin, Germany.
authors.forename: Teng
authors.initials: T
authors.lastname: Huang
authors.affiliation: Institut für Virologie, Freie Universität Berlin, Berlin, Germany.
authors.forename: Nikolaus
authors.initials: N
authors.lastname: Osterrieder
authors.affiliation: Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, New York, United States of America.
authors.forename: James
authors.initials: J
authors.lastname: Catalfamo
mesh_terms: D000022:Abortion, Spontaneous / Q000378:metabolism / Q000821:virology; D000818:Animals; D001792:Blood Platelets / Q000378:metabolism* / Q000821:virology*; D005170:Factor X / Q000378:metabolism; D005260:Female; D006566:Herpesviridae Infections / Q000378:metabolism* / Q000821:virology; D004861:Herpesvirus 1, Equid / Q000378:metabolism*; D006736:Horses / Q000378:metabolism* / Q000821:virology*; D019007:P-Selectin / Q000378:metabolism; D010920:Placenta / Q000378:metabolism / Q000821:virology; D015539:Platelet Activation / Q000502:physiology*; D011247:Pregnancy; D011817:Rabbits; D013917:Thrombin / Q000378:metabolism; D013927:Thrombosis / Q000378:metabolism / Q000821:virology; D014759:Viral Envelope Proteins / Q000378:metabolism
chemical_list: D019007:P-Selectin; D014759:Viral Envelope Proteins; C038535:glycoprotein gC, herpes simplex virus type 1; D005170:Factor X; D013917:Thrombin