Histamine Induces Nuclear Factor of Activated T Cell-Mediated Transcription and Cyclosporin A-Sensitive Interleukin-8 mRNA Expression in Human Umbilical Vein Endothelial Cells
- 1Department of Pharmacology and Program in Molecular Therapeutics and Toxicology, Graduate Division of Biological and Biomedical Sciences (X.W., T.J.M.), 2Emory University School of Medicine, Atlanta, Georgia 30322
Abstract
The nuclear factor of activated T cells (NFAT) mediates a cyclosporin A (CsA)- and FK506-suppressible transcriptional program in lymphocytes after antigen-stimulated phospholipase C activation. Nonlymphoid cells also express NFAT isoforms, raising the possibility that these isoforms can be regulated by other extracellular stimuli. This study sought to determine whether histamine can trigger NFAT-mediated transcription in human umbilical vein endothelial cells (HUVEC), using a retrovirus-based luciferase reporter driven by a well characterized, NFAT-specific enhancer. Luciferase levels are induced up to 60-fold over basal levels after costimulation of HUVEC with Ca2+-mobilizing drugs and a phorbol ester, a response that is 20-fold greater than that observed when HUVEC are stimulated with either drug alone. These synergistic responses are inhibited in cells treated with CsA. CsA and FK506 also inhibit the luciferase response to histamine, indicating that histamine can induce NFAT-mediated transcription in HUVEC. To identify candidate genes in HUVEC that might be regulated by NFAT, the expression of several chemokine mRNAs was measured after histamine treatment. Of the mRNAs tested, only those encoding monocyte chemotactic protein-1 (∼2-fold over basal) and interleukin-8 (∼6-fold over basal) are induced by histamine; both of these responses are suppressed by CsA and FK506. The H1histamine receptor antagonist chlorpheniramine, but not the H2 receptor antagonist ranitidine, blocks the effects of histamine in this preparation. These data provide the first evidence for a physiological inducer of NFAT-mediated transcription in endothelial cells and support the hypothesis that NFAT participates in H1 histamine receptor-induced interleukin-8 gene expression.
Footnotes
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Send reprint requests to: Dr. T. J. Murphy, Department of Pharmacology, 5031 O. W. Rollins Research Building, Emory University School of Medicine, Atlanta, GA 30322. E-mail:tmurphy{at}pharm.emory.edu
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This work was supported by National Institutes of Health Grants HL52810, NS32706, and AR42687. T.J.M. is an Established Investigator of the American Heart Association. An abstract of this work was presented at the Experimental Biology Meeting 1998 (San Francisco, CA).
- Abbreviations:
- NFκB
- nuclear factor κB
- CsA
- cyclosporin A
- NFAT
- nuclear factor of activated T cells
- HUVEC
- human umbilical vein endothelial cell(s)
- MCP-1
- monocyte chemotactic protein-1
- IL
- interleukin
- EGF
- epidermal growth factor
- PDGF-BB
- platelet-derived growth factor BB
- MAP
- mitogen-activated protein
- GM-CSF
- granulocyte/macrophage-colony-stimulating factor
- LTR
- long terminal repeat
- DMEM
- Dulbecco’s modified Eagle medium
- SDS
- sodium dodecyl sulfate
- HBSS
- Hanks’ balanced salt solution
- GAPDH
- glyceraldehyde phosphate dehydrogenase
- PMA
- phorbol myristate acetate
- HEPES
- 4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid
- AP1
- activator protein 1
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- Received January 12, 1998.
- Accepted May 7, 1998.
- The American Society for Pharmacology and Experimental Therapeutics
