3-deazaadenosine inhibits leukocyte adhesion and ICAM-1 biosynthesis in tumor necrosis factor-stimulated human endothelial cells.
Previous reports demonstrate that cultured human umbilical vein endothelial cells (HEC) treated with TNF and other inflammatory mediators show an increased capacity to adhere human neutrophils. This increase is associated with the up-regulation of intercellular adhesion molecule 1 (ICAM-1) and other adhesion molecules on the HEC surface. We have found that 200 microM 3-deazaadenosine (c3Ado) prevented this TNF-induced increase in HEC adhesiveness. This effect resulted from interactions of c3Ado with HEC and not with polymorphonuclear neutrophils. Transport of c3Ado into the HEC was required for its activity, as evidenced by antagonism with the nucleoside transport inhibitor, nitrobenzylthioinosine. Treatment of HEC with c3Ado led to the intracellular buildup of S-adenosylhomocysteine and to the metabolic formation of S-3-deazaadenosylhomocysteine and 3-deazaadenosine 5'-triphosphate, events that appeared not to contribute to c3Ado activity. Exogenous L-homocysteine potentiated c3Ado activity, and this potentiation was prevented by the S-adenosylhomocysteine hydrolase inhibitor, periodate-oxidized adenosine. By using the mAb RR1/1, we have determined that c3Ado also inhibited the TNF-induced expression of ICAM-1 on the surface of the HEC, as well as cytosol-associated ICAM-1. Northern blot and in vitro translation analyses of poly(A+) RNA from c3Ado-treated HEC revealed that this nucleoside analog selectively decreased steady-state levels of ICAM-1 mRNA. The capacity of c3Ado to selectively inhibit HEC adhesiveness, ICAM-1 production, and steady-state levels of ICAM-1 mRNA may contribute to the drug's activity as an anti-inflammatory agent.
Jurgensen
C H
CH
Experimental Therapy Division, Wellcome Research Laboratories, Research Triangle Park, NC 27709.
Huber
B E
BE
Zimmerman
T P
TP
Wolberg
G
G
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents / Q000494:pharmacology*; D015152:Blotting, Northern; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D015815:Cell Adhesion Molecules / Q000096:biosynthesis* / Q000235:genetics; D002462:Cell Membrane / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D015180:Electrophoresis, Gel, Two-Dimensional; D004730:Endothelium, Vascular / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D018799:Intercellular Adhesion Molecule-1; D009504:Neutrophils / Q000166:cytology; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000235:genetics; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D014372:Tubercidin / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*
D000617:Aminoglycosides; D000900:Anti-Bacterial Agents; D015815:Cell Adhesion Molecules; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; C018258:3-deazaadenosine; D018799:Intercellular Adhesion Molecule-1; D012435:S-Adenosylhomocysteine; D014372:Tubercidin