Cytokine Gene Expression in Human LPS- and IFNγ-stimulated Mononuclear Cells Is Inhibited by Heparin

Monica Attanasio; Anna Maria Gori; Betti Giusti; Guglielmina Pepe; Paolo Comeglio; Tamara Brunelli; Domenico Prisco; Rosanna Abbate; Gian Franco Gensini; Gian Gastone Neri Serneri

doi:10.1055/s-0037-1615102

Thrombosis and Haemostasis, Inhaltsverzeichnis

Thromb Haemost 1998; 79(05): 959-962
DOI: 10.1055/s-0037-1615102

Review Article

Schattauer GmbH

Cytokine Gene Expression in Human LPS- and IFNγ-stimulated Mononuclear Cells Is Inhibited by Heparin

Monica Attanasio

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Anna Maria Gori

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Betti Giusti

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Guglielmina Pepe

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

²From the Dipartimento di Biologia, University of Rome “Tor Vergata”, Rome, Italy

,

Paolo Comeglio

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Tamara Brunelli

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Domenico Prisco

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Rosanna Abbate

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Gian Franco Gensini

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

,

Gian Gastone Neri Serneri

¹From the Istituto di Clinica Medica Generale e Cardiologia, University of Florence, Florence

› Institutsangaben

Abstract

Summary

In addition to its well-understood anticoagulant activity, heparin is known to modulate a variety of biological functions including immuno-logic responses. In order to investigate whether heparin influences the humoral immunity by interacting with cellular elements and affecting gene expression in blood circulating cells, we studied the effect of heparin on IL-1β, IL-6 and TNFα mRNAs in human lipopolysaccharide(LPS)- or interferon-γ(IFNγ)-stimulated mononuclear cells. The study of mRNA was carried out by an initial PCR screening followed by a Northern blot quantitative analysis. Heparin (0.5 U/ml) turned out to inhibit all three cytokine gene expressions. The mRNA decrease was 37 ± 6% for IL-1 β, 53 ± 3% for IL-6 and 47 ± 4% for TNF α with LPS stimulus. No differences could be observed in the inhibitory effect of heparin on IFNγ-stimulated cells. This effect of heparin was confirmed in a subset of experiments performed on purified monocytes. These results suggest an important immunosuppressive effect of heparin on cell-mediated immune responses.

Volltext

Referenzen

References
1 Arai K, Lee F, Miyajima A, Miyatake S, Arai N, Yokota T. Cytokines: coordinators of immune and inflammatory responses. Annu Rev Biochem 1990; 59: 783-836.
2 Schindler R, Mancilla J, Endres S, Ghorbani R, Clark SC, Dinarello CA. Correlation and interactions in the production of interleukin-6, IL-1 and TNF in human blood mononuclear cells: IL-6 suppresses IL-1 and TNF. Blood 1990; 75: 40-7.
3 Muller JM, Ziegler-Heitbrock HWL, Baeuerle PA. Nuclear factor kappa B, a mediator of Lipopolysaccharide effects. Immunobiol 1993; 187: 233-56.
4 Blajchman MA, Young E, Ofosu FA. Effects of unfractionated heparin, dermatan sulfate and low molecular weight heparin on vessel wall permeability in rabbits. Ann N Y Acad Sci 1989; 556: 245-54.
5 Clowes AW, Karnowsky MJ. Suppression by heparin of smooth muscle cell proliferation in injured arteries. Nature 1977; 265: 625-6.
6 Castellot Jr JJ, Favreau LV, Karnovsky MJ, Rosemberg RD. Inhibition of vascular smooth muscle cell growth by endothelial cell-derived heparin: possible role of a platelet endoglycosidase. J Biol Chem 1982; 257: 11256-60.
7 Folkman J. Regulation of angiogenesis; a new function of heparin. Biochem Pharmacol 1985; 34: 905-9.
8 Vannucchi S, Pasquali F, Porciatti F, Chiarugi V, Magnelli L, Bianchini P. Binding, internalization and degradation of heparin and heparin fragments by cultured endotelial cells. Thromb Res 1988; 49: 373-83.
9 Glimelius B, Busch C, Hook M. Binding of heparin on the surface of cultured human endothelial cells. Thromb Res 1978; 12 (05) 773-82.
10 Leung L, Saigo K, Grant D. Heparin binds to human monocytes and modulates their pro- coagulant activities and secretory phenotypes. Effect of histidine-rich glycoprotein. Blood 1989; 73: 177-82.
11 Oh TH, Naidoo SS, Jaques LB. The uptake and disposition of S35 heparin by macrophages in vitro. J Reticuloendothel 1973; 13: 134-42.
12 Gallagher JT, Lyon M. Degradation of internalized proteoglycan. In: Heparin, chemical and biological properties, clinical applications. Lane DA, Lindahl U. eds. London: Arnold; 1989. pp 142-3.
13 Au YPT, Kenagy RD, Clowes AW. Heparin selectively inhibits the transcription of tissue-type plasminogen activator in primate arterial smooth muscle cells during mitogenesis. J Biol Chem 1992; 267: 3438-44.
14 Au YPT, Montgomery KF, Clowes AW. Heparin inhibits collagenase gene expression mediated by phorbol ester-responsive element in primate arterial smooth muscle cells. Circulation Res 1992; 70: 1062-9.
15 Wright Jr TC, Pukac LA, Castellot Jr JJ, Karnovsky MJ, Levine RA, Kim-Park HY, Campisi J. Heparin suppresses the induction of c-fos and c-myc mRNA in murine fibroblasts by selective inhibition of a protein kinase C-dependent pathway. Proc Natl Acad Sci USA 1989; 86: 3199-203.
16 Rogers C, Welt FGP, Karnovsky MJ, Edelman ER. Monocyte recruitment and neointimal hyperplasia in rabbits. Coupled inhibitory effects of heparin. Arterioscler Thromb Vasc Biol 1996; 16: 1312-8.
17 Pepe G, Giusti B, Attanasio M, Gori AM, Comeglio P, Martini F, Gensini GF, Abbate R, Neri Serneri GG. Tissue Factor and Plasminogen Activator Inhibitor Type 2 expression in human stimulated monocytes is inhibited by heparin. Semin Thromb Hemost 1997; 23: 135-41.
18 Hogasen AKM, Abrahamsen TG. Heparin suppresses lipopolysaccharide-induced monocyte production of several cytokines, but simultaneously stimulates C3 production. Thromb Res 1995; 80: 179-84.
19 Boyum A. Isolation of lymphocytes, granulocytes and macrophages. Scand J Immunol 1976; 5: 9-15.
20 Bevilacqua MP, Amrani D, Mosesson MW, Bianco C. Receptors for cold-insoluble globulin (plasma fibronectin) on human monocytes. J Exp Med 1981; 153: 42-60.
21 Chomczynski P, Sacchi N. Single-step-method of RNA isolation by acid guanidium thiocyanate-phenol-chloroform extraction. Anal Biochem 1987; 162: 156-9.
22 Feinberg AP, Volgestein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem 1983; 132: 6-11.
23 Farrar MA, Schreiber RD. The molecular cell biology of interferon-γ and its receptor. Annu Rev Immunol 1993; 11: 571-611.
24 Frieri M, Metcalfe DD. Analysis of the effect of mast cell granules on lymphocyte blastogenesis in the absence and presence of mitogens: Identification of heparin as a granule-associated suppressor factor. J Immunol 1983; 131: 1942-7.
25 Currie GA. Effect of heparin on mixed lymphocytes cultures. Nature (Lond.) 1967; 215: 164-9.
26 Ramsden L, Rider C. Selective and differential binding of interleukin (IL)-1a, IL-1b, IL-2 and IL-6 to glycosaminoglycans. Eur J Immunol 1992; 22: 3027-31.
27 Nelson DS. Reaction to antigens in vivo of the peritoneal macrophages of guinea pigs with delayed-type hypersensitivity. Lancet 1963; ii: 175-80.
28 Cohen F, Benacerraf B, McCluskey RT, Ovary Z. Effect of anticoagulants on delayed hypersensitivity reactions. J Immunol 1967; 98: 351-7.
29 Tangelder GJ, Arfors K-E. Inhibition of leukocyte rolling in venules by protamine and sulfated polysaccharides. Blood 1991; 77 (07) 1565-71.
30 Matzner Y, Marx G, Drexler R, Eldor A. The inhibitory effect of heparin and related glycosaminoglycans on neutrophil chemotaxis. Thromb Haemost 1984; 52: 134-7.
31 Nelson MR, Cecconi O, Roberts WG, Aruffo A, Linhardt RJ, Bevilacqua MP. Heparin oligosaccarides bind L- and P-selectin and inhibit acute inflammation. Blood 1993; 82 (11) 3253-8.
32 Zhang Z, Fuentes NL, Fuller GM. Characterization of the IL 6 responsive elements in the γ fibrinogen gene promoter. J Biol Chem 1995; 270: 24287-91.
33 Engelberg H. Low-dose intermittent heparin therapy decreases plasma fibrinogen levels. Semin Thromb Hemost 1991; 17 (Suppl. 02) 219-23.
34 Salvioni A, Marenzi GC, Agostoni P, Grazi S, Guazzi MD. Influence of heparin on fibrinogen and D-dimer plasma levels in acute myocardial infarction treated with streptokinase. Eur Heart J 1994; 15 (05) 654-9.
35 Stouthard J M L, Levi M, Hack E, Veenhof C H N, Romijn HA, Sauerwein HP, van der Poll T. Interleukin-6 stimulates coagulation, not fibrinolysis, in humans. Thromb Haemost 1996; 76 (05) 738-42.