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Interleukin-2 receptor-specific fusion toxin inhibits barotrauma-induced arterial atherosclerosis

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      Abstract

      Immunocytochemical analyses of human plaques and experimental arterial lesions have implicated activated lymphocytes and monocytes in the pathogenesis of atherosclerosis, as demonstrated by the expression of interleukin-2 (IL-2) membrane receptors and major histocompatibility complex class II epitopes. The objective is to determine if targeting these cells with an IL-2 receptor-specific chimeric toxin, DAB486-IL-2, can inhibit experimental post-angioplasty vascular neointimal thickening. Twenty-two atherogenically modeled rabbits were treated in vivo with DAB486-IL-2 (0.1 mg/kg per day i.V.; n = 11) or placebo (n = 11) for 10 days following aortic balloon angioplasty (4 atm × 30 s each × 2 dilatations). In vitro 3H-leucine incorporation studies of mononuclear leukocyte and vascular smooth muscle cell protein synthesis inhibition by DAB486-IL-2 were also performed. Angioplasty sites were examined for evidence of hyperproliferative atherosclerotic narrowing by quantitative angiography and histomorphometry of neointimal cross-sectional area at baseline and 6 weeks after injury. In vitro Concanavalin-A stimulated rabbit mononuclear leukocyte protein synthesis was 50% inhibited by DAB486-IL-2 at a concentration (IC50) of 6 × 10−11 M. Rabbit vascular smooth muscle cells were ~ 150-fold less sensitive to DAB486-IL-2 (IC50 = 10−8 M). In vivo studies showed no change in angioplasty site angiographic minimum luminal diameter at 6 weeks in DAB486-IL-2 treated animals (from 2.96 ± 0.52 to 2.96 ± 0.48 mm; percent cross-sectional area reduction = 1 ± 10%; P = N.S.). In control animals, luminal diameter decreased from 2.79 ± 0.4 to 2.32 ± 0.52 mm at 6 weeks, and percent cross-sectional area was reduced by 34 ± 14% (P < 0.01 vs. placebo). Quantitative histomorphometric angioplasty segmental intimal cross-sectional area reduction of treated and placebo vessels also differed significantly (19 ± 16% vs. 31 ± 21%; P < 0.05). DAB486-IL-2 caused no adverse effects on animal survival, weight or hepatic transaminase levels. We conclude that post-angioplasty administration of the chimeric toxin DAB486-IL-2 inhibits angiographic narrowing and neointimal thickening in the atherogenic rabbit model. Although this IL-2 receptor-specific molecule was cytotoxic in vitro for activated mononuclear leukocytes and vascular smooth muscle cells, systemic toxicity did not occur in vivo at a dose comparable to that evaluated in clinical trials of this agent. Potential anti-proliferative effects of this chimeric toxin may be mediated by direct local inhibition of leukocyte-mediated inflammation, or through the indirect modification of vascular cell mitogenesis and cytokine release.

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      References

        • Ip J.H.
        • Fuster V
        • Badimon L
        • et al.
        Syndromes of accelerated atherosclerosis: roleof vascular injury and smooth cell proliferation.
        J Am Coll Cardiol. 1990; 15: 1667
        • Jonasson L
        • Holm J
        • Hansson G.K.
        Cyclosporin A inhibits smooth muscle proliferation in the vascular response to injury.
        in: 7th Ed. Proc Natl Acad Sci USA. 85. 1988: 2303
        • Berk B.C.
        • Gordon J.B.
        • Alexander R.W.
        Pharmacologic roles of heparin and glucocorticoids to prevent restenosis after coronary angioplasty.
        J Am Coll Cardiol. 1991; 17: 111B
        • Hansson G.K.
        • Jonasson L
        • Lojsthed B
        • et al.
        Localization of T lymphocytes and macrophages in fibrous and complicated human atherosclerotic plaques.
        Atherosclerosis. 1988; 72: 135
        • Hansson G.K.
        • Jonasson L
        • Stemme S
        Immune mechanisms in atherosclerosis.
        Atherosclerosis. 1989; 9: 567
        • Capron L
        • Bruneval P
        Influence of applied stress on mitotic response of arteries to injury with a balloon catheter: quantitative study in rat thoracic aortic.
        Cardiovasc Res. 1989; 23: 941
        • Emeson E.E.
        • Robertson A.L.
        T lymphocytes in aortic and coronary intimas. Their potential role in atherogenesis.
        Am J Pathol. 1988; 130: 369
        • Mitchinson M.J.
        • Ball R.Y.
        Macrophages and atherogenesis.
        The Lancet. 1987; 18: 146
        • Stadius M.L.
        • Rowan R
        • Fleischauer F.J.
        • et al.
        Quantitative histoiogic/immunohistochemical analysis of developing atherosclerotic lesions in previously normal, actuely injured rabbit arteries.
        Circulation. 1990; 82: III
        • Consigny P.M.
        • Tulenko T.N.
        • Nicosia R.F.
        Immediate and long-term effects of angioplasty balloon dilatation on normal rabbit iliac artery.
        Arteriosclerosis. 1986; 6: 265
        • Steele P.M.
        • Chesebro J.H.
        • Stanson A.W.
        • et al.
        Balloon angioplasty in natural history of the pathophysiological response to injury in a pig model.
        Circ Res. 1985; 57: 105
        • Schwartz R.S.
        • Topol E.J.
        • Holmes D.R.
        The restenosis paradigm revisited: an alternative proposal for cellular mechanisms.
        J Am Coll Cardiol. 1992; 20: 1284
        • Weidinger F.F.
        • McLenachan J.M.
        • Cybulsky M.I.
        • Fallon J.T.
        • Hollenberg N.K.
        • Cook J.P.
        • Ganz P
        Hypercholesterolemia enhances macrophage recruitment and dysfunction of regenerated endothelium after balloon injury of the rabbit iliac artery.
        Circulation. 1991; 84: 755
        • Nobuyoshi M
        • Kimura T
        • Oshishi H
        • et al.
        Restenosis after percutaneous coronary angioplasty: pathologic observations in 20 patients.
        J Am Coll Cardiol. 1991; 17: 433
        • Garratt K.N.
        • Edwards W.D.
        • Kaufmann U.P.
        • Vliestra R.E.
        • Holmes D.R.
        Differential histopathology of primary atherosclerosis and restenotic lesions in coronary arteries and saphenous vein grafts: analysis of tissue obtained from 73 patients by direct atherectomy.
        J Am Coll Cardiol. 1991; 17: 442
        • Miller D.D.
        • Craig F.E.
        • Dressier F.A.
        • et al.
        Immunohistochemical characterization of the lymphoid/mononuclear cell composition of human atherectomy tissue: correlation with interleukin-2 receptor expression.
        Atherosclerosis. 1996; (in press)
        • Ip J.H.
        • Fuster V
        • Isreal D
        • Badimon L
        • Badimon J
        • Chesebro J.H.
        The role of platelets, thrombin and hyperplasia in restenosis after coronary angioplasty.
        J Am Coll Cardiol. 1991; 17: 77B
        • Jonasson L
        • Holm J
        • Skalli O
        • Gabliani G
        • Hansson G.K.
        Expression of class 11 transplantation antigen on vascular smooth muscle cells in human atherosclerosis.
        J Clin Invest. 1985; 76: 125
        • Loppnow H
        • Libby P
        Proliferating or interleukin-1 activated human vascular smooth muscle cells seerete copious interleukin-6.
        J Clin Invest. 1990; 85: 731
        • Warner S.J.B.
        • Libby P
        Human vascular smooth muscle cells: target for and source of tumor necrosis factor.
        J Immunol. 1989; 142: 100
        • Jonasson L
        • Holm J
        • Hansson G.K.
        Smooth muscle cells express Ia antigens during arterial response to injury.
        Lab Invest. 1988; 58: 310
        • Casscells W
        Migration of smooth muscle and endothelial cells. Critical events in restenosis.
        Circulation. 1992; 86: 723
        • Stemme S
        • Paterroyo M
        • Hansson G.K.
        Adhesion of activated T-lymphocytes to vascular smooth muscle cells and dermal fibroblasts is mediated by β-1 and β-2 integrins.
        Scand J Immunol. 1992; 36: 233
        • Waldmann T.A.
        The structure, function and expression of interleukin-2 receptors on normal and malignant lymphocytes.
        Science. 1986; 232: 727
        • Clinton S.K.
        • Fleet J.C.
        • Loppnow H
        • et al.
        Interleukin-1 gene expression in rabbit vascular tissue in vivo.
        Am J Pathol. 1991; 138: 1005
        • Hansson G.K.
        • Jonasson L
        • Holm J
        • Clowes M.M.
        • Clowes A.W.
        γ-Interferon regulates vascular smooth muscle proliferation and Ia antigen expression in vivo and in vitro.
        Circ Res. 1988; 63: 712
        • Hollander W
        • Kramsch D.M.
        • Franzblau C
        • Paddock J
        • Colombo M.A.
        Suppression of atheromatous fibrous plaque formation by anti-proliferative and anti-inflammatory drugs.
        Circ. Res. 1974; 34/35: 1
        • Currier J.W.
        • Pow T.K.
        • Minihan A.C.
        • Haudenschild C.C.
        • Faxon D.P.
        • Ryan T.J.
        Colchicine inhibits restenosis after iliac angiopiasty in the atherosclerotic rabbit (abst).
        Circulation. 1989; 80: H
        • Barath P
        • Aaakawa K
        • Cao J
        • et al.
        Low dose of antitumor agents prevents smooth muscle cell proliferation after endothelial injury (abstr).
        J Am Coll Cardiol. 1989; 13: 252A
        • Muller D.W.M.
        • Ellis S.G.
        • Topol E.L
        Colchicine and antineoplastic therapy for the prevention of restenosis after percutaneous coronary interventions.
        J Am Coll Cardiol. 1991; 17: 126B
        • Ferns G
        • Reidy M.A.
        • Ross R
        Vascular effects of cyclosporin A in vivo and in vitro.
        Am J Pathol. 1990; 137: 403
        • Hansson G.K.
        • Holm J
        Interferon-γ inhibits arterial stenosis after injury.
        Circulation. 1991; 84: 1266
        • Murphy J.G.
        • Schwartz R.S.
        • Edwards W.P.
        • et al.
        Methotrexate and azathioprine fail to inhibit porcine coronary restenosis.
        Circulation. 1990; 82: 111
        • O'Keefe J.H.
        • McCallister B.D.
        • Bateman T.M.
        • Kahnlein D
        • Ligon R.W.
        • Hartzler G.O.
        Colchicine for the prevention of restenosis after coronary angioplasty (abst).
        J Am Coll Cardiol. 1991; 17: 181A
        • Pepine C.J.
        • Hirshfeld J.W.
        • MacDonald R.G.
        • et al.
        A controlled trial of corticosteriods to prevent restenosis following angioplasty.
        Circulation. 1990; 81: 1753
        • Casscells W
        • Wai C
        • Shrivastav S
        • et al.
        Smooth muscle proliferation in vessel injury is characterized by expression of fibroblast growth factor receptors and is inhibited by a toxin-fibroblast growth factor conjugate.
        Circulation. 1990; 82: 111
        • Williams D
        • Parker K
        • Bacha P
        • Bishai W
        • Genbauffe F
        • Strom T.B.
        • Murphy J.R.
        Diptheria toxin receptor binding domain substitution with interleukin-1: genetic construction and properties of a diphtheria toxin-related interleukin-2 infusion protein.
        Protein Eng. 1987; 1: 493
        • Bacha P
        • Williams D.P.
        • Waters C
        • Williams J.M.
        • Murphy J.R.
        • Strom The
        Interleukin-2 receptor targeted cytotoxicity: interleukin 2 receptor-mediated action of a diphtheria toxin related interleukin-2 fusion protein.
        J Exp Med. 1988; 167: 612
        • Williams D.P.
        • Wen Z
        • Watson R.S.
        • Boyd J
        • Storms T.B.
        • Murphy J.R.
        Cellular processing of the interleukin-2 fusion toxin DAB468-IL-2 and efficient delivery of diphtheria fragment A to the cytosol of target cells requires Arg194.
        J Biol Chem. 1990; 265: 20673
        • Epstein S.E.A.
        • Siegall C.B.
        • Biro S
        • Fu Y-M
        • Fitzgerald D
        • Pastan I
        Cytotoxic effects of a recombinant chimeric toxin on rapidly proliferating vascular smooth muscle cells.
        Circulation. 1991; 84: 778
        • Biro S
        • Siegall CB
        • Fu Y-M
        • Speir E
        • Pastan I
        • Epstein SE
        In vivo effects of a recombinant toxin targeted to the fibroblast growth factor receptor on rat vascular smooth muscle and endothelial cells.
        Circ Res. 1992; 71: 640
        • Pickering JG
        • Bacha P
        • Weir L
        • Jekanowski J
        • Nichols JC
        • Isner JM
        Prevention of smooth muscle cell outgrowth from human atherosclerotic plaque by a recombinant cytotoxin specific for the epidermal growth factor receptor.
        J Clin Invest. 1993; 91: 724
        • Baumgartner HR
        Eine neue Method zur Erzeugung von Thromben durch zielte Überdehnung der Gefasswand.
        Ges Exp Med. 1963; 173: 227
        • Miller DD
        • Boulet M
        • McEver R
        • et al.
        In vivo S12 monoclonal antibody imaging for detection of plateletmediated mitogen release following atherosclerotic arterial injury.
        Circulation. 1991; 83: 226
        • Waters CA
        • Snider CE
        • Itoh K
        • Poisson L
        • Strom The
        • Murphy JR
        • Nichols JC
        DAB486-IL-2 (IL-2 toxin) selectively inactivates high affinity IL-2 receptor bearing human PBMC. Antigen and clone-specific regulation.
        Ann NY Acad Sci. 1991;
        • Le Maistre CF
        • et al.
        Therapeutic effects of a genetically engineered toxin DAB486-IL-2 in a patient with chronic lymphocytic leukaemia.
        The Lancet. 1991; 337: 1124
        • Sewell KC
        • Parker KC
        • Woodworth TG
        • Reuben J
        • Swartz W
        • Trentham DE
        DAB486-IL-2 fusion toxin in refractory rheumatoid arthritis.
        Arthritis and Rheumatism. 1993; 36: 1223
        • Waters CA
        • Schimke PA
        • Snider CE
        • et al.
        Interleukin-2 receptor targeted cytotoxicity. Receptor requirements for entry of a diptheria toxin-related interleukin-2 fusion protein into cells.
        Eur J Immunol. 1990; 20: 785
        • Gown AM
        • Tsukada T
        • Ross R
        Human atherosclerosis H. Immunocytochemical analysis of the cellular composition of human atherosclerotic lesions.
        Am J Pathol. 1986; 125: 191
        • Weisman AM
        • Hartford JB
        • Svetlik PB
        • Leonard WL
        • Depper JM
        • Waldmann TA
        • Green WG
        • Klausner RD
        Only high-affinity receptors for interleukin-2 mediate internalization of ligant.
        in: 7th Ed. Proc Natl Acad Sci. 83. 1986: 1463
        • Collins L
        • Tsien W-H
        • Seals C
        • Hakim J
        • Weber D
        • Bailon P
        • Hoskings J
        • Greene WC
        • Toome V
        • Ju G
        Identification of specific residues of human interleukin-2 than affect binding to the 70-kDa subunit (p70) of the interleukin-2 receptor.
        in: 7th Ed. Proc Natl Acad Sci USA. 85. 1988: 7709
        • Nabata T
        • Morimoto S
        • Fukuo K
        • Ogihara T
        Biphasic effects of interleukin-2 on proliferation of cultured vascular smooth muscle cells.
        Jap J Pharmacol. 1992; 58 (Abstract): 363P
        • Kagan BL
        • Finkelstein A
        • Colombini M
        Diphtheria toxin fragment forms large pores in phospholipid bilayer membranes.
        in: 7th Ed. Proc Natl Acad Sci. 78. 1981: 4950
        • Murphy JR
        • Strom TB
        Diphtheria toxin-peptide hormone fusion proteins: protein engineering and selective action of a new class of recombitant biological response modifiers.
        in: Moss J Vaughan M ADP-Ribosylating Toxins and G-Proteins: Insights into Signal Transduction. American Society of Microbiology, Washington D.C1990: 141
        • Yamaizwni
        • Mekada E
        • Uchida T
        • Okada Y
        One molecule of diphtheria toxin fragment A introduced into the cell can kill the cell.
        Cell. 1978; 15: 245
        • Miller DD
        • Karim MA
        • Edwards WI
        • Schwartz RS
        Subendothelial lymphoid cell infiltration is a prominent early response to porcine coronary artery injury: relationship to resolution of neointimal fibrin.
        Circulation. 1992; 86: I
        • Reidy MA
        A reassessment of endothelial injury and arterial lesion formation.
        Lab Invest. 1985; 53: 513
        • Prostlewaite AE
        • Holnes MA
        • Katai H
        • Raghow R
        Human fibroblasts synthesize elevated levels of extracellular matrix proteins in response to interleukin-4.
        J Clin Invest. 1992; 90: 1497
        • Stehbens WE
        An appraisal of cholesterol feeding in experimental atherogenesis.
        Prog Cardiovasc Dis. 1986; 29: 107
        • Ferrell M
        • Fuster V
        • Gold HK
        • Chesebro JH
        A dilemma for the 1990's. Choosing appropriate experimental animal model for the prevention of restenosis.
        Circulation. 1992; 85: 1630
        • Sarembock U
        • Gertz SD
        • Gimple LW
        • Owen RM
        • Powders E
        • Roberts WC
        Effectiveness of recombinant desulphatohirudin in reducing restenosis after balloon angioplasty of atherosclerotic femoral arteries in rabbits.
        Circulation. 1991; 84: 232
        • Hansson GK
        • Holm J
        • Jonasson L
        Detection of activated T lymphocytes in the human atherosclerotic plaque.
        Am J Pathol. 1989; 135: 165
        • Bieber CP
        • et al.
        Malignant lymphoma in cyclosporin A treated allograft recipients.
        Lancet. 1980; 1: 43
        • Olisari MT
        • Kubo SH
        • Braunlin EA
        • et al.
        Five-year experience with triple-drug immunosuppressive therapy in cardiac transplantation.
        Circulation. 1990; 82: 276