Suppression of human monocyte tissue factor synthesis by antisense oligodeoxynucleotide.
Tissue factor (TF) is a cellular receptor and cofactor for factor V11/V11a which initiates the blood coagulation cascade. An understanding of TF cell biology is therefore of critical importance to the pathophysiology of many disorders. We have utilized an antisense oligodeoxynucleotide (aODN), directed against human blood monocyte TF mRNA, to investigate the feasibility of inhibiting the synthesis of TF. CD14 receptor-mediated endocytosis was used as a means of delivering TF aODN to monocytes. This DNA carrier system consists of the fab portion of anti-CD14 antibody covalently linked to poly (L-lysine). Co-treatment of monocytes with TF aODN and endotoxin resulted in 80.4 +/- 2.2% suppression of TF activity when compared with control endotoxin stimulated cells. Control experiments with TF sense ODN, mismatched aODN, and an irrelevant aODN were performed to exclude nonspecific inhibitory effects. The cytotoxicity of the DNA carrier complex was also evaluated. These results demonstrate that this TF mRNA antisense ODN specifically suppressed the synthesis of biologically active monocyte TF and that antisense ODNs might therefore represent a useful tool in the investigation of monocyte/macrophage TF function.
Stephens
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Department of Paediatrics, Imperial College School of Medicine at St. Mary's, London, UK.
Rivers
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RP
D015152:Blotting, Northern; D005167:Factor VII / Q000378:metabolism; D015942:Factor VIIa / Q000378:metabolism; D015951:Factor Xa / Q000378:metabolism; D006801:Humans; D009000:Monocytes / Q000378:metabolism*; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D013873:Thionucleotides / Q000494:pharmacology*; D013925:Thromboplastin / Q000096:biosynthesis* / Q000235:genetics
D016376:Oligonucleotides, Antisense; D012333:RNA, Messenger; D013873:Thionucleotides; D005167:Factor VII; D013925:Thromboplastin; D015942:Factor VIIa; D015951:Factor Xa