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Inhibition of endothelial cell adhesion molecule expression with antisense oligonucleotides.

C F Bennett, T P Condon, S Grimm, H Chan and M Y Chiang
J Immunol April 1, 1994, 152 (7) 3530-3540;
C F Bennett
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T P Condon
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S Grimm
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H Chan
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M Y Chiang
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Abstract

In response to inflammatory stimuli, expression of a group of proteins that bind circulating leukocytes (endothelial-leukocyte adhesion molecules) are induced on the luminal surface of vascular endothelium. A series of phosphorothioate oligonucleotides 18 to 21 bases in length were designed and synthesized to hybridize selectively to the mRNA, which encodes three such endothelial-leukocyte adhesion molecules; human intercellular adhesion molecule-1 (ICAM-1), vascular cell adhesion molecule-1 (VCAM-1), and E-selectin. Antisense oligonucleotides were identified that selectively inhibited ICAM-1, VCAM-1, and E-selectin expression in HUVEC. Oligonucleotides that hybridized to the 3'-untranslated region of either ICAM-1, VCAM-1, or E-selectin mRNAs promoted a selective reduction in the respective mRNA levels. In contrast, oligonucleotides that hybridized to 5'-untranslated sequences did not significantly reduce target mRNA levels, although they did promote a reduction in protein expression. With the use of flow cytometry to measure cell surface expression, ICAM-1 and E-selectin were selectively inhibited by their respective antisense oligonucleotide. At low concentrations of oligonucleotides, only VCAM-1 antisense oligonucleotides inhibited VCAM-1 expression. However, at an oligonucleotide concentration of 50 nM or greater, phosphorothioate oligonucleotides not predicted to hybridize to VCAM-1 mRNA also reduced VCAM-1 expression. The sequence-independent inhibition of VCAM-1 expression by phosphorothioate oligonucleotides could be the result of a perturbation in the transcriptional regulation of the VCAM-1 gene. ICAM-1, VCAM-1, and E-selectin antisense oligonucleotides reduced adhesion of HL-60 cells to TNF-activated HUVEC. These data demonstrate that phosphorothioate oligonucleotides are capable of selectively inhibiting the expression of ICAM-1, VCAM-1, and E-selectin in HUVEC.

  • Copyright © 1994 by American Association of Immunologists

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The Journal of Immunology
Vol. 152, Issue 7
1 Apr 1994
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Inhibition of endothelial cell adhesion molecule expression with antisense oligonucleotides.
C F Bennett, T P Condon, S Grimm, H Chan, M Y Chiang
The Journal of Immunology April 1, 1994, 152 (7) 3530-3540;

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Inhibition of endothelial cell adhesion molecule expression with antisense oligonucleotides.
C F Bennett, T P Condon, S Grimm, H Chan, M Y Chiang
The Journal of Immunology April 1, 1994, 152 (7) 3530-3540;
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Print ISSN 0022-1767        Online ISSN 1550-6606