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(Arteriosclerosis, Thrombosis, and Vascular Biology. 1997;17:423-428.)
© 1997 American Heart Association, Inc.

Articles

Early Activation Signals in Endothelial Cells

Stimulation by Cytokines

Martino Introna; ; Alberto Mantovani

From the Dipartimento di Immunologia e Biologia Cellulare, Istituto di Ricerche Farmacologiche "Mario Negri," via Eritrea 62, 20157 Milano (M.I., A.M.), and Sezione di Patologia Generale, Università di Brescia, Brescia (A.M.), Italy.

Correspondence to Martino Introna, MD, Laboratory of Molecular Immunohematology, Department of Immunology and Cell Biology, Istituto di Ricerche Farmacologiche "Mario Negri," via Eritrea 62, 20157 Milano, Italy. E-mail Martino{at}IRFMN.MNEGRI.IT

	Abstract

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
Abstract With limitation to the "proinflammatory program" induced in endothelial cells by exposure to interleukin-1, tumor necrosis factor, and interleukin-6, we review the available data on the signaling for these three cytokines, from receptor engagement to induction of gene transcription. Only a few molecular pathways have been characterized so far, and key issues in endothelial biology, such as endothelial specificity of gene expression and heterogeneity of different endothelial populations, remain largely unexplored.

Key Words: endothelial cells • cytokines • molecular signaling • gene expression

	Introduction

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
In the past several years, the concept that ECs, rather than being a passive barrier between blood elements and tissues, represent a metabolically active organ has been consolidated. In particular, emphasis has been placed on the role played by ECs in determining and maintaining the inflammatory reactions that occur in tissues.^{1 2 3 4} All of the crucial aspects that characterize inflammation, from the release of chemoattractants for different leukocyte populations to the expression and functional activation of adhesion molecules of different classes and from the induction of prothrombotic activity on the luminal surface to the transmigration and functional activation of the motile elements, are mainly determined by the early metabolic response of ECs to inflammatory signals.^{1 2 3 4}

To organize this vast amount of information, several years ago we defined the complex series of molecular events that occur in ECs upon exposure to the classic proinflammatory cytokines (eg, IL-1 and TNF) as the induction of the "prothrombotic-proinflammatory program."^{1 4} In this review, we will concentrate on only selected, recent aspects of molecular events that take place in ECs and limit our discussion to IL-1, TNF, and IL-6 stimulation as paradigms of proinflammatory cytokines. Furthermore, we will not recapitulate the list of the many known genes or activities that have been found to be "induced" upon activation but rather concentrate on what has recently been learned about the mechanisms whereby some of these genes or activities are induced. In particular, this review will consider only those genes whose transcriptional regulation has been directly studied in ECs. Finally, in agreement with the scope of this review, the discussion will focus on those aspects that appear to be the most promising for future investigation.

	Receptors and Associated Proteins

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
ECs express both the p55 and the p75 TNFR. After its binding to TNF, p75 is internalized and transported to the lysosomes via coated pits and coated vesicles, whereas p55 proceeds mainly to the Golgi apparatus and cytoplasmic vacuoles.⁵ Most studies agree on the predominance of the p55 molecule (ie, TNFR I) for signaling,^{6 7 8} but recently a specific role for p75 (TNFR II) has been proposed, since it has been shown to be engaged by membrane TNF⁹ and therefore could be involved in signaling after cell-cell interactions.^{9 10 11 12}

Although much information has been accumulated on molecules that physically associate with the two TNFRs, particularly for signaling the apoptotic response (for a review, see References 13¹³ and 14¹⁴ ) in other cell systems, no such data on the molecular interactions that exist in ECs after TNFR occupancy by TNF are available.

ECs express the type I IL-1 receptor only,¹⁵ a property peculiar to perhaps this cell type in that most other cell types tested so far also possess variable amounts of the "decoy" type II IL-1 receptor.¹⁶ So far as signaling or receptor-associated molecules are concerned, the IIL-1RAcP, which cooperates functionally in IL-1 signaling, is expressed at low levels in ECs (S. Saccani et al, unpublished results, 1997).

Original studies on IL-6 showed that this cytokine did not affect several EC functions,¹⁷ a conclusion repeatedly confirmed despite indications for its involvement in angiogenesis and vascular tumor formation. Observations in IL-6–knockout mice prompted us to reexamine IL-6 interactions with ECs, and we found that ECs express the gp130 chain but not the IL-6 R-chain. Furthermore, soluble IL-6 R-chain alone and IL-6 alone did not affect EC function, but the two together induced chemokine production in ECs (M. Romano et al, in press).

	Signaling

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
Few reports have explored the role of tyrosine phosphorylation in the cytokine activation of ECs. It has been reported that the TNF-induced signal-transduction pathway leading to plasminogen activator inhibitor-1 transcription involves a genistein-sensitive step that is not involved in the induction of urokinase-type plasminogen activator by TNF.¹⁸ Whereas a role for tyrosine phosphorylation has been demonstrated for different cytokines in ECs (References 19¹⁹ and 20²⁰ and below), it was also recently claimed for two adhesion molecules (VCAM-1 and E-selectin) that are induced by TNF.²¹

In ECs TNF has been shown to activate the three MAP kinases: p42ERK, p38, and p54JNK, which are the most downstream effectors of the MAP kinase cascade.²² Additional data point out JNK in particular, which leads to ATF-2/c-Jun heterodimer activation.²³ This complex, together with CREB proteins and ATF-2 homodimers, plays a role in the transcription of the E-selectin promoter^{24 25} and cooperates with NF-B on this promoter (see below). All members of this group belong to the basic-Leu zipper family of transcription factors and require phosphorylation for functional activation and subsequent dimerization. Previous studies had shown that pharmacologically relevant elevations of cAMP levels in ECs perturbed E-selectin expression,²⁶ and a role for phosphokinase A–mediated phosphorylation had been hypothesized in studies of forskolin-mediated activation of ECs.²⁷ More recent studies have shown that cAMP decreases ATF-2/c-Jun heterodimers while increasing ATF-2 homodimers, an action opposite to that of TNF.²³ It is known that each of three MAP kinases requires the phosphorylation of at least two other kinases farther upstream in the cascade, MAPKK and MAPKKK. However, the link between the TNFR and activation of the first kinase in each cascade in ECs is not yet understood.

Recently, a role for ceramide has been hypothesized in the activation of Raf-1 (one of the MAPKKKs) that occurs after TNF stimulation and leads to activation of ERK.²² Ceramide is known to be a product of activation of the "sphingomyelin pathway" (hydrolysis of sphingomyelin to ceramide). In this case also, however, the molecular mechanism by which the TNF/IL-1 receptors activate this pathway in EC has yet to be unraveled. Moreover, ceramide does not seem to be sufficient to induce a biological response like adhesion molecule expression, but it can amplify induction by IL-1.^{22 28} Ceramide may have some cooperative role in the canonical NF-B activation induced by IL-1 and TNF (see below), but it appears to lead to the formation of complexes (p50/p65 heterodimers) different from those induced by IL-1 in the same cells (p50 homodimers or p50/other subunit heterodimers as well as p50/p65 heterodimers).²⁸ Interestingly, a hypothesis for hematopoietic cells has recently been proposed, which suggests that two different sphingomyelinases act in a different way, the neutral enzyme in the plasma membrane being responsible for the ceramide production that leads to MAP kinase activation, and the acidic one in the lysosomes producing the ceramide (through intermediate diacylglycerol production) responsible for NF-B activation.²⁹ The FAN protein (factor associated with neutral sphingomyelinase activation) has been cloned; it couples the p55 TNFR with neutral sphingomyelinase.³⁰ Whether different pathways of ceramide production are also present in ECs and which role these may play in these cells should therefore become an interesting subject of investigation.

The JaK/STAT pathway is only now being explored in ECs. In particular, after previous reports that IL-4 induced VCAM-1 expression through tyrosine phosphorylation of several cellular substrates (including phosphorylation of its own receptor¹⁹ ), more recently IL-13 has also been shown to activate a tyrosine kinase that phosphorylates the 140-kD IL-4 R chain³¹ ; furthermore, both cytokines activate JaK2 phosphorylation and the STAT6 transcription factor,³¹ although activation of JaK 3 has also been reported in ECs after IL-4 stimulation.³² Indeed, IL-4 and IL-13 have been repeatedly reported to induce similar biological responses^{4 17} in ECs and may therefore share some receptor component, although ECs have been found to be negative for the -chain, the common chain of IL-4, -2, -7, -9, and -15 receptors,^{31 33} thus implying that some other molecular component is shared between IL-4 and IL-13 receptors in ECs. We very recently observed that granulocyte macrophage-colony stimulating factor, which induces expression of a functional program in ECs related to angiogenesis,³⁴ is also able to activate JaK2 kinase through a physical association with the ß-chain of the receptor (R. Soldi et al, unpublished data).

Finally, following the demonstration that IL-6/IL-6 R complexes could activate ECs (see above), we observed STAT3 phosphorylation under the same experimental conditions (X. Romano et al, in press). These observations may now pave the way for more extensive characterization of the role played by different members of the JaK and STAT families in the different metabolic programs that are inducible in ECs.^{1 4}

	Activation of NF-B

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
Despite initial conflicting results, it is now clear that an increase in NF-B activity follows IL-1/TNF stimulation of ECs. Indeed, "activated" NF-B activity has been detected in ECs in atherosclerotic plaques in vivo.³⁵ NF-B activation takes place by posttranscriptional modification, and two major mechanisms are likely to play a role in its activation, namely, phosphorylation of the inhibitory molecule IB and free radical–dependent oxidation. It is well established that NF-B elements are tightly controlled by several inhibitors belonging to the IB family.³⁶ In ECs, it has been demonstrated that TNF exposure induces Ser phosphorylation and subsequent proteolytic degradation of IB-³⁷ ; very recently, two kinases of 36 and 41 kD have been identified in the cytoplasm of ECs that specifically bind to and phosphorylate the IB- subunit.³⁸ Similarly, several Ser protease inhibitors have been shown to inhibit IB- degradation and to consequently block TNF-induced upregulation of adhesion molecules.³⁹ More recently, persistent activation of NF-B has been observed in ECs after rapid and persistent degradation of IB-ß.⁴⁰ These last results do not explain why the more classic TNF/IL-1–inducible genes show only a transient induction of expression. As strong evidence for the role played by IB-, ectopic expression of this molecule in ECs induces marked inhibition of several molecules, such as VCAM-1; IL-1, -6, and -8; and tissue factor.⁴¹

Interestingly, many data suggest a role for reactive oxygen intermediates as common and critical denominators for various activating signals leading to NF-B activation. In particular, many different antioxidants, such as NAC, DTCs, vitamin E derivatives, metal chelators, and pyrrolidine DTCs, have been shown to effectively block NF-B activation and subsequent gene induction. This issue has been scarcely explored in ECs: -Tocopherol was shown to inhibit cytokine-induced expression of E-selectin, but the authors failed to detect any change in NF-B activity on the promoter.⁴² On the contrary, other agents like pyrrolidine DTC and NAC were shown to inhibit expression of VCAM-1, possibly through a reduction in the binding of NF-B proteins to promoter elements, but interestingly in the same study, no effect was reported with respect to E-selectin or ICAM.⁴³ More recently, pyrrolidine DTC has been shown able to abrogate TF expression after activation by different cytokines. including IL-1 and TNF.⁴⁴

All in all, proteolytic degradation of IB plays a pivotal role in NF-B activation in ECs, and the necessity of the proteosome pathway has been documented, showing functional inhibition of TNF-induced EC activation by peptide aldehyde inhibitors of the proteosome.⁴⁵

	Inhibition of NF-B

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
After cell activation while IB- protein levels fall rapidly,^{37 46} its RNA levels are dramatically upregulated, since it is the gene under NF-B transcriptional control. The newly made protein should then resequester NF-B in the cytoplasm. Interestingly, IB- was found among the IL-1 "immediate early inducible" genes cloned from IL-1–stimulated ECs and described to be rapidly upregulated even in the absence of protein synthesis.^{47 48 49} The circuit of IB protein degradation followed by IB gene induction may provide a model for the transient induction of many cytokine-inducible genes. More recently, regulated expression of the NF-B and IB- system has been demonstrated in vivo in an animal model of arterial injury.⁵⁰

Yet another protein, the zinc finger transcription factor A20, originally cloned as an "immediate early" response gene in TNF-stimulated ECs⁵¹ that was also shown to protect cells from TNF-induced cytotoxicity,⁵² has recently been shown to inhibit NF-B activity in ECs⁵³ by a mechanism that has yet to be explored.

	Which NF-B Complex?

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
NF-B refers to a family of transcription factors that dimerize with various affinities for the consensus sequence GGGRNNYYCC. The canonical NF-B complex consists of a p65/p50 heterodimer, although a certain EC specificity is emerging for the p65/c-Rel heterodimer (see below). Furthermore, many genes important to EC biology contain variant NF-B HGGRNNYYCC binding sites (see below). Data are also accumulating on the "fine tuning" that regulates the different compositions of NF-B complexes after EC activation. One example has already been given of the different and opposite effects of cAMP and TNF on the relative concentrations of the dimers.⁵⁴ Another example suggests that the EC specificity of VCAM-1 expression may be related to the type of complex induced by TNF. Indeed, these authors showed that the canonical p65/p50 heterodimer does not activate the VCAM-1 promoter, possibly due to negative interference of p50. Furthermore, they suggest that dimers containing p65 but not p50 are mainly responsible for activation and suggest that formation of these still incompletely characterized active dimers may be specific for ECs.⁵⁵ A schematic representation of IL-1, TNF, and IL-6 signaling in ECs is shown in the Figure.

View larger version (30K): [in this window] [in a new window]   Figure 1. IL-1, TNF, and IL-6 signaling in ECs. TNF receptors are dimers; each monomer is characterized by four immunoglobulin-like domains. TNF is active mainly as a trimer. IL-1RI also belongs to the immunoglobulin superfamily. The extracellular region of the IL-6R chain comprises one immunoglobulin-like domain and the cytokine receptor family module. The extracellular region of gp130 comprises four fibronectin type III domains and one cytokine receptor family module. Signaling is described in the text. ROI indicates reactive oxygen intermediates.

	Gene Promoters

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
To date, the best characterized promoter in ECs is that for the E-selectin gene. Canonical p50/p65 NF-B heterodimers are able to bind to three distinct sites in the E-selectin promoter,^{24 56 57} now identified as PDI, PDIII, and PDIV.⁵⁸ An additional element is a CRE/ATF–like binding site that has recently been localized within PDII and binds a phosphorylated ATF-2 dimer (see above). The latter functionally cooperates with NF-B activity in stimulated ECs.^{25 59} Finally, in all four binding sites, the presence of the high-mobility group protein I has also been described, which may⁶⁰ facilitate the binding of NF-B and ATF factors. In general, the overall organization of the promoter is very reminiscent of the interferon-beta promoter, and a similar stereospecific model has therefore been hypothesized.⁵⁸

Characterization of the VCAM-1 promoter has shown the presence of tandem NF-B elements, both of which are necessary for cytokine-mediated expression and are bound by classic p65/p50 heterodimers or p65 homodimers.^{58 61 62 63} More recently, other elements have been described, including one IRF-1 and one Sp-1 binding site.⁶⁴ Finally, the high-mobility group protein appears to facilitate NF-B and IRF-1 binding and seems to be a crucial element for cytokine inducibility of the gene.⁶⁵ More recently, an upstream element has been invoked as a potential negative regulator that could function as an IL-1–dependent transcriptional repressor (and does not respond to TNF) in human dermal microvascular ECs but be inactive in other ECs, such as human umbilical vein ECs.⁶⁶

The ICAM-1 promoter offers yet another example of cooperation between different factors on proximal sites: one atypical NF-B site mediates binding of p50/p65 heterodimers, c-Rel/p65, and p65 homodimers,^{67 68} and one -activated sequence element mediates binding to p91/ STAT1 in response to interferon gamma.⁶⁹ The functional role of this element has been highlighted in epithelial cells, whereas the modest induction of ICAM-1 in ECs by interferon gamma seems to imply the presence of tissue-specific negative regulatory factors.⁷⁰ Finally, one c/EBP element mediates binding to several factors (ie, C/EBP- and -ß).⁵⁸ Study of the interactions between different factors (as in the case of NF-B and c/EBP) could reveal additional elements responsible for ICAM-1 inducibility by different signals (eg, one AP-1 element and the ets transcription factor appear to be involved during ICAM-1 induction by exposure to H₂O₂⁷¹ ). Characterization of the tissue factor promoter has also begun. TNF inducibility seems to require cooperation between two AP-1 binding sites and one NF-B element^{44 72 73} and possibly also the Sp-1⁷⁴ factor. As for ICAM-1, this NF-B element is a variant sequence that is able to mediate c-Rel/p65 binding.⁶⁸

	Another Level of Integration

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
The important functional role described for CREB in TNF-mediated E-selectin transcription (see above) opens new questions that may be of great interest in future investigations. It is now known that the large adaptor molecule CBP and the closely related p300 can bind to phosphorylated CREB as well as to many other transcription factors, such as c-myb, AP-1, nuclear receptors, and others.⁷⁵ More importantly, it has been shown that CBP is a limiting factor in cells and may serve as an integrator of multiple signal transduction pathways within the cell.⁷⁵ Since several transcription factors have been demonstrated to play a role in ECs (see above) and may compete with CBP, it will be interesting to explore whether such a mechanism plays a role, for example, when ECs are stimulated by cytokines that induce very different metabolic programs.

	Conclusions

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
All in all, we have discussed that many molecules have now been identified at different levels and that they may all play a role in cytokine interactions on the EC surface and/or the ensuing biological response, but complete molecular pathways have yet to be characterized. Furthermore, key biological questions remain largely unexplored, such as the endothelial specificity of certain genes or the nature of EC heterogenicity, and we have indicated the few reports that address these questions. Finally, the plasticity shown by ECs in their diverse biological responses after stimulation with different groups of cytokines^{2 4} may require the systematic cloning and identification of other genes, possibly by the "subtracting/differential" techniques of screening.^{47 52 76}

	Selected Abbreviations and Acronyms

 

ATF = activating transcription factor c/EBP = CCAAT enhancer binding protein CBP = CREB binding protein CREB = cAMP responsive element binding EC(s) = endothelial cell(s) ERK = extracellular regulated kinase gp = glycoprotein IL = interleukin IRF = interferon regulatory factor JaK = Janus kinase JNK = jun NH2-terminal kinase KK(K) = kinase kinase (kinase) MAP = mitogen-activated protein NAC = N-acetyl-L-cysteine PD = positive regulatory domain PKA = protein kinase A R = receptor RAcP = receptor-associated protein STAT = signal transduction and activator of transcription TNF = tumor necrosis factor VCAM = vascular cell adhesion molecule

	Acknowledgments

 
This work was supported in part by grants provided by Consiglio Nazionale delle Ricerche (94;02828;CT04 to A.M.), by Istituto Superiore di Sanità (AIDS project to A.M.), and by the Italian Association for Cancer Research (to M.I.).

Received December 6, 1996; accepted December 18, 1996.

	References

Top Abstract Introduction Receptors and Associated... Signaling Activation of NF-{kappa}B Inhibition of NF-{kappa}B Which NF-{kappa}B Complex? Gene Promoters Another Level of Integration Conclusions References

 
1. Mantovani A, Dejana E. Cytokines as communication signals between leukocytes and endothelial cells. Immunol Today. 1989;10:370-375.[Medline] [Order article via Infotrieve]

2. Pober J, Cotran RS. Cytokines and endothelial cell biology. Physiol Rev. 1990;70:427-451.[Free Full Text]

3. Libby P, Hansson GK. Biology of disease: involvement of the immune system in human atherogenesis—current knowledge and unanswered questions. Lab Invest. 1991;64:5-15.[Medline] [Order article via Infotrieve]

4. Mantovani A, Bussolino F, Dejana E. Cytokine regulation of endothelial cell function. FASEB J. 1992;6:2591-2599.[Abstract]

5. Bradley JR, Thiru S, Pober JS. Disparate localization of 55-kd and 75-kd tumor necrosis factor receptors in human endothelial cells. Am J Pathol. 1995;146:27-32.[Abstract]

6. Mackay F, Loetscher H, Stueber D, Gehr G, Lesslauer W. Tumor necrosis factor alpha (TNF-alpha)-induced cell adhesion to human endothelial cells is under dominant control of one TNF receptor type, TNF-R55. J Exp Med. 1993;177:1277-1286.[Abstract/Free Full Text]

7. Paleolog EM, Delasalle SA, Buurman WA, Feldmann M. Functional activities of receptors for tumor necrosis factor-alpha on human vascular endothelial cells. Blood. 1994;84:2578-2590.[Abstract/Free Full Text]

8. Slowik MR, De Luca LG, Fiers W, Pober JS. Tumor necrosis factor activates human endothelial cells through the p55 tumor necrosis factor receptor but the p75 receptor contributes to activation at low tumor necrosis factor concentration. Am J Pathol. 1993;143:1724-1730.[Abstract]

9. Grell M, Douni E, Wajant H, Löheden M, Clauss M, Maxeiner B, Georgopoulos S, Lesslauer W, Kollias G, Pfizenmaier K, Scheurich P. The transmembrane form of tumor necrosis factor is the prime activating ligand of the 80 kDa tumor necrosis factor receptor. Cell. 1995;83:793-802.[Medline] [Order article via Infotrieve]

10. Tartaglia LA, Pennica D, Goeddel DV. Ligand passing: the 75-kDa tumor necrosis factor (TNF) receptor recruits TNF for signaling by the 55-kDa TNF receptor. J Biol Chem. 1993;268:18542-18548.[Abstract/Free Full Text]

11. Schmid EF, Binder K, Grell M, Scheurich P, Pfizenmaier K. Both tumor necrosis factor receptors, TNFR60 and TNFR80, are involved in signaling endothelial tissue factor expression by juxtacrine tumor necrosis factor alpha. Blood. 1995;86:1836-1841.[Abstract/Free Full Text]

12. Lukacs NW, Strieter RM, Elner V, Evanoff HL, Burdick MD, Kunkel SL. Production of chemokines, interleukin-8 and monocyte chemoattractant protein-1, during monocyte endothelial cell interactions. Blood. 1995;86:2767-2773.[Abstract/Free Full Text]

13. Fraser A, Evan G. A license to kill. Cell. 1996;85:781-784.[Medline] [Order article via Infotrieve]

14. Bazzoni F, Beutler B. The tumor necrosis factor ligand and receptor families. N Engl J Med. 1996;334:1717-1725.[Free Full Text]

15. Colotta F, Sironi M, Borre A, Pollicino T, Bernasconi S, Boraschi D, Mantovani A. Type II interleukin-1 receptor is not expressed in cultured endothelial cells and is not involved in endothelial cell activation. Blood. 1993;81:1347-1351.[Abstract/Free Full Text]

16. Colotta F, Re F, Muzio M, Bertini R, Polentarutti N, Sironi M, Giri JG, Dower SK, Sims JE, Mantovani A. Interleukin-1 type II receptor: a decoy target for IL-1 that is regulated by IL-4. Science. 1993;261:472-475.[Abstract/Free Full Text]

17. Sironi M, Breviario F, Proserpio P, Biondi A, Vecchi A, Van Damme J, Dejana E, Mantovani A. IL-1 stimulates IL-6 production in endothelial cells. J Immunol. 1989;142:549-553.[Abstract]

18. van Hinsbergh VW, Vermeer M, Koolwijk P, Grimbergen J, Kooistra T. Genistein reduces tumor necrosis factor alpha-induced plasminogen activator inhibitor-1 transcription but not urokinase expression in human endothelial cells. Blood. 1994;84:2984-2991.[Abstract/Free Full Text]

19. Palmer-Crocker RL, Pober JS. IL-4 induction of VCAM-1 on endothelial cells involves activation of a protein tyrosine kinase. J Immunol. 1995;154:2838-2845.[Abstract]

20. Schieven GL, Kallestad JC, Brown TJ, Ledbetter JA, Linsley PS. Oncostatin M induces tyrosine phosphorylation in endothelial cells and activation of p62yes tyrosine kinase. J Immunol. 1992;149:1676-1682.[Abstract]

21. Weber C, Negrescu E, Erl W, Pietsch A, Frankenberger M, Ziegler Heitbrock HW, Siess W, Weber PC. Inhibitors of protein tyrosine kinase suppress TNF-stimulated induction of endothelial cell adhesion molecules. J Immunol. 1995;155:445-451.[Abstract]

22. Modur V, Zimmerman GA, Prescott SM, McIntyre TM. Endothelial cell inflammatory responses to tumor necrosis factor alpha: ceramide-dependent and -independent mitogen-activated protein kinase cascades. J Biol Chem. 1996;271:13094-13102.[Abstract/Free Full Text]

23. De Luca LG, Johnson DR, Whitley MZ, Tucker C, Pober JS. cAMP and tumor necrosis factor competitively regulate transcriptional activation through and nuclear factor binding to the cAMP-responsive element/activating transcription factor element of the endothelial leukocyte adhesion molecule-1 (E-selectin) promoter. J Biol Chem. 1994;269:19193-19196.[Abstract/Free Full Text]

24. Whitley MZ, Thanos D, Read MA, Maniatis T, Collins T. A striking similarity in the organization of the E-selectin and beta interferon gene promoters. Mol Cell Biol. 1994;14:6464-6475.[Abstract/Free Full Text]

25. Kaszubska W, van Huijsduijnen RH, Ghersa P, DeRaemy Schenk AM, Chen BP, Hai T, DeLamarter JF, Whelan J. Cyclic AMP-independent ATF family members interact with NF-kappa B and function in the activation of the E-selectin promoter in response to cytokines. Mol Cell Biol. 1993;13:7180-7190.[Abstract/Free Full Text]

26. Pober JS, Slowik MR, DeLuca LG, Ritchie AJ. Elevated cyclic AMP inhibits endothelial cell synthesis and expression of TNF-induced endothelial leukocyte adhesion molecule-1, and vascular cell adhesion molecule-1, but not intercellular adhesion molecule-1. J Immunol. 1993;150:5114-5123.[Abstract]

27. Ghersa P, Hooft van Huijsduijnen R, Whelan J, Cambet Y, Pescini R, DeLamarter JF. Inhibition of E-selectin gene transcription through a cAMP-dependent protein kinase pathway. J Biol Chem. 1994;269:29129-29137.[Abstract/Free Full Text]

28. Masamune A, Igarashi Y, Hakomori S. Regulatory role of ceramide in interleukin (IL)-1 beta-induced E-selectin expression in human umbilical vein endothelial cells: ceramide enhances IL-1 beta action, but is not sufficient for E-selectin expression. J Biol Chem. 1996;271:9368-9375.[Abstract/Free Full Text]

29. Wiegmann K, Schütze S, Machleidt T, Witte D, Krönke M. Functional dichotomy of neutral and acidic sphingomyelinases in tumor necrosis factor signaling. Cell. 1994;78:1005-1015.[Medline] [Order article via Infotrieve]

30. Adam-Klages S, Adam D, Wiegmann K, Struve S, Kolanus W, Schneider-Mergener J, Krönke M. FAN, a novel WD-repeat protein couples the p55 TNF-receptor to neutral sphingomyelinase. Cell. 1996;86:937-947.[Medline] [Order article via Infotrieve]

31. Palmer-Crocker RL, Hugles CCW, Pober JS. IL-4 and IL-13 activate the JAK2 tyrosine kinase and Stat6 in cultured human vascular endothelial cells through a common pathway that does not involve the gammaC chain. J Clin Invest. 1996;98:604-609.[Medline] [Order article via Infotrieve]

32. Verbsky JW, Bach EA, Fang YF, Yang L, Randolph DA, Fields LE. Expression of Janus kinase 3 in human endothelial and other non-lymphoid and non-myeloid cells. J Biol Chem. 1996;271:13976-13980.[Abstract/Free Full Text]

33. Schnyder B, Lugli S, Feng N, Etter H, Lutz RA, Ryffel B, Sugamura K, Wunderli-Allenspach H, Moser R. IL-4 and IL-13 bind to a shared heterodimeric complex on endothelial cells mediating vascular cell adhesion molecule-1 induction in the absence of the common gamma chain (gamma-c). Blood. 1996;87:4286-4295.[Abstract/Free Full Text]

34. Bussolino F, Bocchietto E, Silvagno F, Soldi R, Arese M, Mantovani A. Actions of molecules which regulate hemopoiesis on endothelial cells: memoirs of common ancestors? Pathol Res Pract. 1994;190:834-839.[Medline] [Order article via Infotrieve]

35. Brand K, Page S, Rogler G, Bartsch A, Brandl R, Knuechel R, Page M, Kaltschmidt C, Baeuerle PA, Neumeier D. Activated transcription factor nuclear factor-kappa B is present in the atherosclerotic lesion. J Clin Invest. 1996;97:1715-1722.[Medline] [Order article via Infotrieve]

36. Miyaloto S, Maki M, Schmitt MJ, Hatanaka M, Verna IM. Tumor necrosis factor alpha-induced phosphorylation of IkBalpha is signal for its degradation but not dissociation from NF-kB. Proc Natl Acad Sci U S A. 1994;91:12740-12744.[Abstract/Free Full Text]

37. Read MA, Whitley MZ, Williams AJ, Collins T. NF-kappa B and I kappa B alpha: an inducible regulatory system in endothelial activation. J Exp Med. 1994;179:503-512.[Abstract/Free Full Text]

38. Bennett BL, Lacson RG, Chen CC, Cruz R, Wheeler JS, Kletzien RF, Tomasselli AG, Heinrikson RL, Manning AM. Identification of signal-induced IkB- kinases in human endothelial cells. J Biol Chem. 1996;271:19680-19688.[Abstract/Free Full Text]

39. Chen CC, Rosenbloom CL, Anderson DC, Manning AM. Selective inhibition of E-selectin, vascular cell adhesion molecule-1, and intercellular adhesion molecule-1 expression by inhibitors of I kappa B-alpha phosphorylation. J Immunol. 1995;155:3538-3545.[Abstract]

40. Johnson DR, Douglas I, Jahnke A, Ghosh S, Pober JS. A sustained reduction in IKappaB-beta may contribute to persistent NF-kappaB activation in human endothelial cells. J Biol Chem. 1996;271:16317-16322.[Abstract/Free Full Text]

41. Wrighton CJ, Hofer Warbinek R, Moll T, Eytner R, Bach FH, de Martin R. Inhibition of endothelial cell activation by adenovirus-mediated expression of IkappaBalpha, an inhibitor of the transcription factor NF-kappaB. J Exp Med. 1996;183:1013-1022.[Abstract/Free Full Text]

42. Faruqi R, de la Motte C, DiCorleto PE. Alpha-tocopherol inhibits agonist-induced monocytic cell adhesion to cultured human endothelial cells. J Clin Invest. 1994;94:592-600.

43. Marui N, Offermann MK, Swerlick R, Kunsch C, Rosen CA, Ahmad M, Alexander RW, Medford RM. Vascular cell adhesion molecule-1 (VCAM-1) gene transcription and expression are regulated through an antioxidant-sensitive mechanism in human vascular endothelial cells. J Clin Invest. 1993;92:1866-1874.

44. Orthner CL, Rodgers GM, Fitzgerald LA. Pyrrolidine dithiocarbamate abrogates tissue factor (TF) expression by endothelial cells: evidence implicating nuclear factor-kappa B in TF induction by diverse agonists. Blood. 1995;86:436-443.[Abstract/Free Full Text]

45. Read MA, Neish AS, Luscinskas FW, Palombella VJ, Maniatis T, Collins T. The proteosome pathway is required for cytokine-induced endothelial-leukocytes adhesion molecule expression. Immunity. 1995;2:493-506.[Medline] [Order article via Infotrieve]

46. de Martin R, Vanhove B, Cheng Q, Hofer E, Csizmadia V, Winkler H, Bach FH. Cytokine-inducible expression in endothelial cells of an I kappa B alpha-like gene is regulated by NF kappa B. EMBO J. 1993;12:2773-2779.[Medline] [Order article via Infotrieve]

47. Breviario F, d'Aniello EM, Golay J, Peri G, Bottazzi B, Bairoch A, Saccone S, Marzella R, Predazzi V, Rocchi M, Della Valle G, Dejana E, Mantovani A, Introna M. Interleukin-1-inducible genes in endothelial cells: cloning of a new gene related to C-reactive protein and serum amyloid P component. J Biol Chem. 1992;267:22190-22197.[Abstract/Free Full Text]

48. Groves RW, Giri J, Sims J, Dower SK, Kupper TS. Inducible expression of type 2 IL-1 receptors by cultured human keratinocytes: implication for IL-1-mediated processes in epidermis. J Immunol. 1995; 154:4065-4072.

49. d'Aniello EM, Breviario F, Martin Padura I, Lampugnani MG, Dejana E, Mantovani A, Introna M. Interleukin-1 and tumor necrosis factor induce transient expression of an inhibitor of nuclear factor kB in endothelial cells. Endothelium. 1993;1:161-165.

50. Lindner V, Collins T. Expression of NF-kB and IkB-alpha by aortic endothelium in an arterial injury model. Am J Pathol. 1996;148:427-438.[Abstract]

51. Dixit VM, Green S, Sarma V, Holzman LB, Wolf FW, O'Rourke K, Ward PA, Prochownik EV, Marks RM. Tumor necrosis factor-alpha induction of novel gene products in human endothelial cells including a macrophage-specific chemotaxin. J Biol Chem. 1990;265:2973-2978.[Abstract/Free Full Text]

52. Opipari AWJ, Hu HM, Yabkowitz R, Dixit VM. The A20 zinc finger protein protects cells from tumor necrosis factor cytotoxicity. J Biol Chem. 1992;267:12424-12427.[Abstract/Free Full Text]

53. Cooper JT, Stroka DM, Brostjan C, Palmetshofer A, Bach FH, Ferran C. A20 blocks endothelial cell activation through a NF-kappaB-dependent mechanism. J Biol Chem. 1996;271:18068-18073.[Abstract/Free Full Text]

54. Miyagishi R, Kikuchi S, Fukazawa T, Tashiro K. Macrophage inflammatory protein-1 alpha in the cerebrospinal fluid of patients with multiple sclerosis and other inflammatory neurological diseases. J Neurol Sci. 1995;129:223-227.[Medline] [Order article via Infotrieve]

55. Ahmad M, Marui N, Alexander RW, Medford RM. Cell type-specific transactivation of the VCAM-1 promoter through an NF-kappa B enhancer motif. J Biol Chem. 1995;270:8976-8983.[Abstract/Free Full Text]

56. Schindler U, Baichwal VR. Three NF-kappa B binding sites in the human E-selectin gene required for maximal tumor necrosis factor alpha-induced expression. Mol Cell Biol. 1994;14:5820-5831.[Abstract/Free Full Text]

57. Hooft van Huijsduijnen R, Pescini R, DeLamarter JF. Two distinct NF-kappa B complexes differing in their larger subunit bind the E-selectin promoter kappa B element. Nucleic Acids Res. 1993;21:3711-3717.[Abstract/Free Full Text]

58. Collins T, Read MA, Neish AS, Whitley MZ, Thanos D, Maniatis T. Transcriptional regulation of endothelial cell adhesion molecules: NF-kappa B and cytokine-inducible enhancers. FASEB J. 1995;9:899-909.[Abstract]

59. Pescini R, Kaszubska W, Whelan J, DeLamarter JF, Hooft van Huijsduijnen R. ATF-a0, a novel variant of the ATF/CREB transcription factor family, forms a dominant transcription inhibitor in ATF-a heterodimers. J Biol Chem. 1994;269:1159-1165.[Abstract/Free Full Text]

60. Lewis H, Kaszubska W, DeLamarter JF, Whelan J. Cooperativity between two NF-kappa B complexes, mediated by high-mobility-group protein I(Y), is essential for cytokine-induced expression of the E-selectin promoter. Mol Cell Biol. 1994;14:5701-5709.[Abstract/Free Full Text]

61. Neish AS, Williams AJ, Palmer HJ, Whitley MZ, Collins T. Functional analysis of the human vascular cell adhesion molecule 1 promoter. J Exp Med. 1992;176:1583-1593.[Abstract/Free Full Text]

62. Iademarco MF, McQuillan JJ, Rosen GD, Dean DC. Characterization of the promoter for vascular cell adhesion molecule-1 (VCAM-1). J Biol Chem. 1992;267:16323-16329.[Abstract/Free Full Text]

63. Shu HB, Agranoff AB, Nabel EG, Leung K, Duckett CS, Neish AS, Collins T, Nabel GJ. Differential regulation of vascular cell adhesion molecule 1 gene expression by specific NF-kappa B subunits in endothelial and epithelial cells. Mol Cell Biol. 1993;13:6283-6289.[Abstract/Free Full Text]

64. Neish AS, Khachigian LM, Park A, Baichwal VR, Collins T. Sp1 is a component of the cytokine-inducible enhancer in the promoter of vascular cell adhesion molecule-1. J Biol Chem. 1995;270:28903-28909.[Abstract/Free Full Text]

65. Neish AS, Read MA, Thanos D, Pine R, Maniatis T, Collins T. Endothelial interferon regulatory factor 1 cooperates with NF-kB as a transcriptional activator of vascular cell adhesion molecule 1. Mol Cell Biol. 1995;15:2558-2569.[Abstract]

66. Gille J, Swerlick RA, Lawley TJ, Caughman SW. Differential regulation of vascular cell adhesion molecule-1 gene transcription by tumor necrosis factor alpha and interleukin-1 alpha in dermal microvascular endothelial cells. Blood. 1996;87:211-217.[Abstract/Free Full Text]

67. Ledebur HC, Parks TP. Transcriptional regulation of the intercellular adhesion molecule-1 gene by inflammatory cytokines in human endothelial cells: essential roles of a variant NF-kappa B site and p65 homodimers. J Biol Chem. 1995;270:933-943.[Abstract/Free Full Text]

68. Parry GC, Mackman N. A set of inducible genes expressed by activated human monocytic and endothelial cells contain kappa B-like sites that specifically bind c-Rel-p65 heterodimers. J Biol Chem. 1994;269:20823-20825.[Abstract/Free Full Text]

69. Hou J, Baichwal V, Cao Z. Regulatory elements and transcription factors controlling basal and cytokine-induced expression of the gene encoding intercellular adhesion molecule 1. Proc Natl Acad Sci U S A. 1994;91:11641-11645.[Abstract/Free Full Text]

70. Look DC, Pelletier MR, Holtzman MJ. Selective interaction of a subset of interferon-gamma response element-binding proteins with the intercellular adhesion molecule-1 (ICAM-1) gene promoter controls the pattern of expression on epithelial cells. J Biol Chem. 1994;269:8952-8958.[Abstract/Free Full Text]

71. Roebuck KA, Rahman A, Lakshminarayanan V, Janakidevi K, Malik AB. H2O2 and tumor necrosis factor-alpha activate intercellular adhesion molecule 1 (ICAM-1) gene transcription through distinct cis-regulatory elements within the ICAM-1 promoter. J Biol Chem. 1995;270:18966-18974.[Abstract/Free Full Text]

72. Bierhaus A, Zhang Y, Deng Y, Mackman N, Quehenberger P, Haase M, Luther T, Muller M, Bohrer H, Greten J, et al. Mechanism of the tumor necrosis factor alpha-mediated induction of endothelial tissue factor. J Biol Chem. 1995;270:26419-26432.[Abstract/Free Full Text]

73. Mackman N. Regulation of the tissue factor gene. FASEB J. 1995;9:883-889.[Abstract]

74. Moll T, Czyz M, Holzmuller H, Hofer Warbinek R, Wagner E, Winkler H, Bach FH, Hofer E. Regulation of the tissue factor promoter in endothelial cells: binding of NF kappa B-, AP-1-, and Sp1-like transcription factors. J Biol Chem. 1995;270:3849-3857.[Abstract/Free Full Text]

75. Janknecht R, Hunter T. A growing coactivator network. Nature. 1996;383:22-23.[Medline] [Order article via Infotrieve]

76. Chu W, Burns DK, Swerlick RA, Presky DH. Identification and characterization of a novel cytokine-inducible nuclear protein from human endothelial cells. J Biol Chem. 1995;270:10236-10245.[Abstract/Free Full Text]

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