Subcutaneous Administration of Low-Molecular-Weight Heparin to Horses Inhibits Ex Vivo Equine Herpesvirus Type 1-Induced Platelet Activation.
Abstract: Equine herpesvirus type 1 (EHV-1) is a major cause of infectious respiratory disease, abortion and neurologic disease. Thrombosis in placental and spinal vessels and subsequent ischemic injury in EHV-1-infected horses manifests clinically as abortion and myeloencephalopathy. We have previously shown that addition of heparin anticoagulants to equine platelet-rich plasma (PRP) can abolish ex vivo EHV-1-induced platelet activation. The goal of this study was to test whether platelets isolated from horses treated with unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) were resistant to ex vivo EHV-1-induced activation. In a masked, block-randomized placebo-controlled cross-over trial, 9 healthy adult horses received 4 subcutaneous injections at q. 12 h intervals of one of the following treatments: UFH (100 U/kg loading dose, 3 maintenance doses of 80 U/kg), 2 doses of LMWH (enoxaparin) 80 U/kg 24 h apart with saline at the intervening 12 h intervals, or 4 doses of saline. Blood samples were collected before treatment and after 36 h, 40 h (4 h after the last injection) and 60 h (24 h after the last injection). Two strains of EHV-1, Ab4 and RacL11, were added to PRP ex vivo and platelet membrane expression of P selectin was measured as a marker of platelet activation. Drug concentrations were monitored in a Factor Xa inhibition (anti-Xa) bioassay. We found that LMWH, but not UFH, inhibited platelet activation induced by low concentrations (1 × 106 plaque forming units/mL) of both EHV-1 strains at 40 h. At this time point, all horses had anti-Xa activities above 0.1 U/ml (range 0.15-0.48 U/ml) with LMWH, but not UFH. By 60 h, a platelet inhibitory effect was no longer detected and anti-Xa activity had decreased (range 0.03 to 0.07 U/ml) in LMWH-treated horses. Neither heparin inhibited platelet activation induced by high concentrations (5 × 106 plaque forming units/mL) of the RacL11 strain. We found substantial between horse variability in EHV-1-induced platelet activation at baseline and after treatment. Minor injection site reactions developed in horses given either heparin. These results suggest that LMWH therapy may prevent thrombotic sequelae of EHV-1, however further evaluation of dosage regimens is required.
Publication Date: 2018-05-28 PubMed ID: 29892605PubMed Central: PMC5985713DOI: 10.3389/fvets.2018.00106Google Scholar: Lookup
The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.
The research investigates the effect of low-molecular-weight heparin (LMWH) on horses infected with the equine herpesvirus type 1 (EHV-1), finding that administering LMWH can help inhibit the virus-induced activation of platelets, potentially preventing the serious thrombotic consequences of the infection. Further evaluations of dosage regimens are needed.
Study Design and Goals
- The study aimed to determine whether platelets taken from horses treated with unfractionated heparin (UFH) or low-molecular-weight heparin (LMWH) become resistant to the activation induced by EHV-1.
- Nine healthy adult horses were injected subcutaneously with either UFH, LMWH, or saline at regular intervals.
- Heparin is an anticoagulant previously shown to inhibit EHV-1-induced platelet activation.
Testing and Measurements
- Blood samples were collected before treatment and 36, 40, and 60 hours after treatment.
- Two strains of EHV-1 were added to the equine platelet-rich plasma (PRP), and platelet activation was measured through the presence of P selectin in the platelet membrane.
- The researchers also monitored drug concentrations through a Factor Xa inhibition assay (anti-Xa).
Results Summary
- The study found that LMWH, but not UFH, was able to inhibit platelet activation caused by low concentrations of the EHV-1 strains at the 40-hour mark. At this time, higher than normal anti-Xa activities were also registered in the LMWH group but not UFH.
- By 60 hours, the platelet inhibitory effect was no longer detected, and anti-Xa activity had also decreased in the LMWH group.
- Neither type of heparin was able to inhibit platelet activation induced by high concentrations of one of the strains, RacL11.
Conclusion and Implications
- The results demonstrate a significant variability between horses in terms of EHV-1-induced platelet activation both at baseline and post-treatment.
- Minor reactions were noted at the injection site in horses given either type of heparin.
- These findings suggest that LMWH might help prevent the thrombotic effects of EHV-1 infection and calls for more research to fine-tune the dosage regimen.
Cite This Article
APA
Stokol T, Serpa PBS, Brooks MB, Divers T, Ness S.
(2018).
Subcutaneous Administration of Low-Molecular-Weight Heparin to Horses Inhibits Ex Vivo Equine Herpesvirus Type 1-Induced Platelet Activation.
Front Vet Sci, 5, 106.
https://doi.org/10.3389/fvets.2018.00106 Publication
Researcher Affiliations
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
- Department of Population Medicine and Diagnostic Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
- Department of Clinical Sciences, College of Veterinary Medicine, Cornell University, Ithaca, NY, United States.
References
This article includes 36 references
- Holz CL, Nelli RK, Wilson ME, Zarski LM, Azab W, Baumgardner R. Equine disease surveillance: quarterly summary.. Vet Rec (2016) 179(5):115–8.
- Anagha G, Gulati BR, Riyesh T, Virmani N. Genetic characterization of equine herpesvirus 1 isolates from abortion outbreaks in India.. Arch Virol (2017) 162(1):157–63.
- Damiani AM, de Vries M, Reimers G, Winkler S, Osterrieder N. A severe equine herpesvirus type 1 (EHV-1) abortion outbreak caused by a neuropathogenic strain at a breeding farm in northern Germany.. Vet Microbiol (2014) 172(3-4):555–62.
- Walter J, Seeh C, Fey K, Bleul U, Osterrieder N. Clinical observations and management of a severe equine herpesvirus type 1 outbreak with abortion and encephalomyelitis.. Acta Vet Scand (2013) 55:19.
- Barbić L, Lojkić I, Stevanović V, Bedeković T, Starešina V, Lemo N. Two outbreaks of neuropathogenic equine herpesvirus type 1 with breed-dependent clinical signs.. Vet Rec (2012) 170(9):227.
- Goehring LS, van Winden SC, van Maanen C, Sloet van Oldruitenborgh-Oosterbaan MM. Equine herpesvirus type 1-associated myeloencephalopathy in The Netherlands: a four-year retrospective study (1999-2003).. Journal of Veterinary Internal Medicine / American College of Veterinary Internal Medicine (2006) 20(3):601–7.
- Lunn DP, Davis-Poynter N, Flaminio MJ, Horohov DW, Osterrieder K, Pusterla N. Equine herpesvirus-1 consensus statement.. J Vet Intern Med (2009) 23(3):450–61.
- Wilsterman S, Soboll-Hussey G, Lunn DP, Ashton LV, Callan RJ, Hussey SB. Equine herpesvirus-1 infected peripheral blood mononuclear cell subpopulations during viremia.. Vet Microbiol (2011) 149(1-2):40–7.
- Smith DJ, Iqbal J, Purewal A, Hamblin AS, Edington N. In vitro reactivation of latent equid herpesvirus-1 from CD5+/CD8+ leukocytes indirectly by IL-2 or chorionic gonadotrophin.. J Gen Virol (1998) 79 ( Pt 12):2997–3004.
- Smith DJ, Hamblin AS, Edington N. Infection of endothelial cells with equine herpesvirus-1 (EHV-1) occurs where there is activation of putative adhesion molecules: a mechanism for transfer of virus.. Equine Vet J (2001) 33(2):138–42.
- Goehring LS, Hussey GS, Ashton LV, Schenkel AR, Lunn DP. Infection of central nervous system endothelial cells by cell-associated EHV-1.. Vet Microbiol (2011) 148(2-4):389–95.
- Edington N, Bridges CG, Patel JR. Endothelial cell infection and thrombosis in paralysis caused by equid herpesvirus-1: equine stroke.. Arch Virol (1986) 90(1-2):111–24.
- Edington N, Smyth B, Griffiths L. The role of endothelial cell infection in the endometrium, placenta and foetus of equid herpesvirus 1 (EHV-1) abortions.. J Comp Pathol (1991) 104(4):379–87.
- Smith KC, Mumford JA, Lakhani K. A comparison of equid herpesvirus-1 (EHV-1) vascular lesions in the early versus late pregnant equine uterus.. J Comp Pathol (1996) 114(3):231–47.
- Goehring LS, Soboll Hussey G, Gomez Diez M, Benedict K, Maxwell LK, Morley PS. Plasma D-dimer concentrations during experimental EHV-1 infection of horses.. J Vet Intern Med (2013) 27(6):1535–42.
- Yeo WM, Osterrieder N, Stokol T. Equine herpesvirus type 1 infection induces procoagulant activity in equine monocytes.. Vet Res (2013) 44(1):16.
- Stokol T, Yeo WM, Burnett D, Deangelis N, Huang T, Osterrieder N. Equid herpesvirus type 1 activates platelets.. PLoS One (2015) 10(4):e0122640.
- Pryzdial EL, Sutherland MR, Ruf W. The procoagulant envelope virus surface: contribution to enhanced infection.. Thromb Res (2014) 133 Suppl 1(Suppl 1):S15–S17.
- Feige K, Schwarzwald CC, Bombeli T. Comparison of unfractioned and low molecular weight heparin for prophylaxis of coagulopathies in 52 horses with colic: a randomised double-blind clinical trial.. Equine Vet J (2003) 35(5):506–13.
- de La Rebière de Pouyade G, Grulke S, Detilleux J, Salciccia A, Verwilghen DR, Caudron I. Evaluation of low-molecular-weight heparin for the prevention of equine laminitis after colic surgery.. J Vet Emerg Crit Care (2009) 19(1):113–9.
- Gerhards H, Kietzmann M. Dose-dependent plasma elimination of subcutaneously administered calcium heparin in horses.. J Vet Pharmacol Ther (1988) 11(1):77–83.
- Gerhards H, Eberhardt C. Plasma heparin values and hemostasis in equids after subcutaneous administration of low-dose calcium heparin.. Am J Vet Res (1988) 49(1):13–18.
- Schwarzwald CC, Feige K, Wunderli-Allenspach H, Braun U. Comparison of pharmacokinetic variables for two low-molecular-weight heparins after subcutaneous administration of a single dose to horses.. Am J Vet Res (2002) 63(6):868–73.
- Whelchel DD, Tennent-Brown BS, Giguère S, Epstein KL. Pharmacodynamics of multi-dose low molecular weight heparin in healthy horses.. Vet Surg (2013) 42(4):448–54.
- Duncan SG, Meyers KM, Reed SM. Reduction of the red blood cell mass of horses: toxic effect of heparin anticoagulant therapy.. Am J Vet Res (1983) 44(12):2271–6.
- Monreal L, Villatoro AJ, Monreal M, Espada Y, Anglés AM, Ruiz-Gopegui R. Comparison of the effects of low-molecular-weight and unfractioned heparin in horses.. Am J Vet Res (1995) 56(10):1281–5.
- Moore JN, Mahaffey EA, Zboran M. Heparin-induced agglutination of erythrocytes in horses.. Am J Vet Res (1987) 48(1):68–71.
- Gerhards H. Low dose calcium heparin in horses: plasma heparin concentrations, effects on red blood cell mass and on coagulation variables.. Equine Vet J (1991) 23(1):37–43.
- Epstein KL. Coagulopathies in horses.. Vet Clin North Am Equine Pract (2014) 30(2):437–52.
- Stokol T, Serpa PBS, Zahid MN, Brooks MB. Unfractionated and Low-Molecular-Weight Heparin and the Phosphodiesterase Inhibitors, IBMX and Cilostazol, Block Ex Vivo Equid Herpesvirus Type-1-Induced Platelet Activation.. Front Vet Sci (2016) 3:99.
- Maxwell LK, Bentz BG, Gilliam LL, Ritchey JW, Pusterla N, Eberle R. Efficacy of the early administration of valacyclovir hydrochloride for the treatment of neuropathogenic equine herpesvirus type-1 infection in horses.. Am J Vet Res (2017) 78(10):1126–39.
- Walter J, Seeh C, Fey K, Bleul U, Osterrieder N. Prevention of equine herpesvirus myeloencephalopathy - Is heparin a novel option? A case report.. Tierarztl Prax Ausg G Grosstiere Nutztiere (2016) 44(5):313–7.
- Scruggs JL, Flatland B, Mccormick KA, Reed A. Biological variation of thromboelastrography variables in 10 clinically healthy horses.. J Vet Emerg Crit Care (2016) 26(1):80–4.
- Rudez G, Meijer P, Spronk HM, Leebeek FW, Ten Cate H, Kluft C. Biological variation in inflammatory and hemostatic markers.. J Thromb Haemost (2009) 7(8):1247–55.
- Goodman LB, Loregian A, Perkins GA, Nugent J, Buckles EL, Mercorelli B. A point mutation in a herpesvirus polymerase determines neuropathogenicity.. PLoS Pathog (2007) 3(11):e160.
- Holz CL, Nelli RK, Wilson ME, Zarski LM, Azab W, Baumgardner R. Viral genes and cellular markers associated with neurological complications during herpesvirus infections.. J Gen Virol (2017) 98(6):1439–54.
Citations
This article has been cited 3 times.- Theuerkauf K, Obach-Schröck C, Staszyk C, Moritz A, Roscher KA. Activated platelets and platelet-leukocyte aggregates in the equine systemic inflammatory response syndrome. J Vet Diagn Invest 2022 May;34(3):448-457.
- Huang L, Chen G, Hu B, Liang S, Chu W, Chen L. Preventive application of low molecular weight heparin ameliorates peripherally inserted central catheter-related venous thrombosis. Int J Clin Exp Pathol 2020;13(3):403-410.
- Serpa PBS, Brooks MB, Divers T, Ness S, Birschmann I, Papich MG, Stokol T. Pharmacokinetics and Pharmacodynamics of an Oral Formulation of Apixaban in Horses After Oral and Intravenous Administration. Front Vet Sci 2018;5:304.
Use Nutrition Calculator
Check if your horse's diet meets their nutrition requirements with our easy-to-use tool Check your horse's diet with our easy-to-use tool
Talk to a Nutritionist
Discuss your horse's feeding plan with our experts over a free phone consultation Discuss your horse's diet over a phone consultation
Submit Diet Evaluation
Get a customized feeding plan for your horse formulated by our equine nutritionists Get a custom feeding plan formulated by our nutritionists
