Generation and characterization of a monoclonal antibody against canine tissue factor

Stokol, Tracy; DeLeonardis, Christine; Dong, Lynn; Wagner, Bettina

doi:10.1016/j.vetimm.2015.07.001

Cited by 1 publication

(2 citation statements)

References 24 publications

(46 reference statements)

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“…Ideally, this test would have been performed directly by inhibition of canine TF with specific antibodies. 45 We previously tested several inhibitory anti-human TF antibodies including a monoclonal murine antibody (anti-human TF1) and a murine anti-canine TF monoclonal antibody generated in our own laboratory, 31 but none inhibited canine TF procoagulant activity in the CAT assay. The activity of FVIIa is enhanced 5,000-fold by binding of TF and membrane surfaces, which results in an allosteric change in FVIIa structure.…”

Section: Discussionmentioning

confidence: 99%

“…Detached cells (1 × 10 5 cells/reaction) were re-suspended in PBS with 1% bovine serum albumin d and 0.05% sodium azide (ie, reaction buffer) and were incubated with monoclonal mouse anti-dog TF antibody IgG (clone 133-2) 31 or mouse IgG isotype control q (each used at a final concentration of 20 μg/mL) for 30 minutes on ice. After washing in reaction buffer, a secondary fluorescent-conjugated goat anti-mouse IgG r (1:200 dilution) was added, followed by incubation for 30 minutes on ice.…”

Section: Methodsmentioning

confidence: 99%

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Evaluation of procoagulant tissue factor expression in canine hemangiosarcoma cell lines

Witter¹,

Gruber²,

Lean³

et al. 2017

ajvr

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OBJECTIVE To evaluate expression of procoagulant tissue factor (TF) by canine hemangiosarcoma cells in vitro. SAMPLES 4 canine hemangiosarcoma cell lines (SB-HSA [mouse-passaged cutaneous tumor], Emma [primary metastatic brain tumor], and Frog and Dal-1 [primary splenic tumors]) and 1 nonneoplastic canine endothelial cell line (CnAoEC). PROCEDURES TF mRNA and TF antigen expression were evaluated by quantitative real-time PCR assay and flow cytometry, respectively. Thrombin generation was measured in canine plasma and in coagulation factor–replete or specific coagulation factor–deficient human plasma by calibrated automated thrombography. Corn trypsin inhibitor and annexin V were used to examine contributions of contact activation and membrane-bound phosphatidylserine, respectively, to thrombin generation. RESULTS All cell lines expressed TF mRNA and antigen, with significantly greater expression of both products in SB-HSA and Emma cells than in CnAoEC. A greater percentage of SB-HSA cells expressed TF antigen, compared with other hemangiosarcoma cell lines. All hemangiosarcoma cell lines generated significantly more thrombin than did CnAoEC in canine or factor-replete human plasma. Thrombin generation induced by SB-HSA cells was significantly lower in factor VII-deficient plasma than in factor-replete plasma and was abolished in factor X–deficient plasma; residual thrombin generation in FVII-deficient plasma was abolished by incubation of cells with annexin V. Thrombin generation by SB-HSA cells was unaffected by the addition of corn trypsin inhibitor. CONCLUSIONS AND CLINICAL RELEVANCE Hemangiosarcoma cell lines expressed procoagulant TF in vitro. Further research is needed to determine whether TF can be used as a biomarker for hemostatic dysfunction in dogs with hemangiosarcoma.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%