RESUMEN
OBJECTIVE: To determine the efficacy of soluble pig tissue factor pathway inhibitor fusion immunoglobulin (TFPI-Ig) in blocking pig to human xenogeneic blood coagulation. RESULTS: To generate pig TFPI-Ig or human TFPI-Ig, expression vector containing cDNA encoding pig TFPIα or human TFPIα combined with human constant Ig heavy chain region was cloned and introduced into CHO cells. After purification of pig TFPI-Ig and human TFPI-Ig, the inhibition of each recombinant protein on pig tissue factor (TF)-mediated blood coagulation was examined in human plasma. Compared to human TFPI-Ig, pig TFPI-Ig inhibited pig TF activity and thrombin generation in human plasma more efficiently at certain concentrations. CONCLUSIONS: Pig TFPI-Ig will be be useful as a therapeutic protein to treat pig to human xenogeneic blood coagulation.
Asunto(s)
Inmunoglobulinas/genética , Lipoproteínas/genética , Plasma/química , Proteínas Recombinantes/farmacología , Tromboplastina/antagonistas & inhibidores , Animales , Coagulación Sanguínea/efectos de los fármacos , Células CHO , Cricetulus , Células HeLa , Humanos , Inmunoglobulinas/metabolismo , Lipoproteínas/metabolismo , PorcinosRESUMEN
The CD90 (Thy-1) is a glycosylphosphatidylinositol (GPI)-anchored glycoprotein that transfers signals involved in many biological events including cell activation, cell migration, cell adhesion, and tumor suppression. In this study, we cloned pig CD90 cDNA and determined its complete cDNA sequence. Pig CD90 cDNA contained an open reading frame (486 bp) encoding 161 amino acids with three putative N-glycosylation sites and four well-conserved cysteine residues, which form a possible disulfide bond within the extracellular domain among mammalian species. Pig CD90 mRNA was detected in various tissues, indicating the multicellular functions of CD90 in pigs. Flow cytometry analyses demonstrated that anti-human CD90 antibody recognizes a pig CD90 on the cell surface. Moreover, immunohistochemistry analysis revealed that CD90 expression is widely diffused in several pig tissues. Further studies will be necessary to define the functional contribution of CD90 during specific infectious diseases in pigs.