Research paper| Volume 10, ISSUE 2, P130-138, September 1995

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The effects of cyclosporin A and FK506 on proliferation and IL-8 production of cultured human keratinocytes

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      The present study was conducted to determine whether cyclosporin A (CsA) and FK506 could be effective in inhibiting the proliferation and cytokine secretion of normal human epidermal keratinocytes (NHEK). NHEK proliferation in the presence of CsA and FK506 at the concentrations 10−9 to 10−5 M at 24 and 48 h time points was measured colorimetrically by the MTS assay. CsA had inhibitory effects from 10−6 to 10−5 M, while FK506 had no effect, except for toxicity at the very highest concentrations (5 × 10−6 M and higher). NHEK cells spontaneously secrete IL-8 (243.4 ± 55.5 pg/ml), and this baseline level was augmented by TNF-α alone, or synergistically by TNF-α and IFN-γ, which are thought to be secreted by T cells. Neither CsA nor FK506 had any significant effect on either spontaneous or cytokine-stimulated keratinocyte IL-8 production. Therefore, it is most likely that the two drugs indirectly inhibit the keratinocyte inflammatory response through their actions on T cells or other immunocompetent cells.



      MTS ((3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethophenyl)-2-(4-sulfophenyl)-2H-tetrazolium) or Owen's reagent), PMS (phenazine methosulfate)
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        • Cohen DJ
        • Loertscher R
        • Rubin MF
        • Tilney NL
        • Carpenter CB
        • Strom TB
        Cyclosporine: a new immunosuppressive agent for organ transplantation.
        Ann Int Med. 1984; 101: 667-682
        • Tugwell P
        • Bombardier C
        • Gent M
        • et al.
        Low-dose cyclosporin versus placebo in patients with rheumatoid arthritis.
        Lancet. 1990; 335: 1051-1055
        • Ellis CN
        • Gorsulowsky DC
        • Hamilton TA
        • et al.
        Cyclosporine improves psoriasis in a double-blind study.
        J Am Med Assoc. 1986; 256: 3110-3116
        • Ellis CN
        • Fradin MS
        • Messana JM
        • et al.
        Cyclosporine for plaque-type psoriasis. Results of a multidose, double- blind trial.
        N Engl J Med. 1991; 324: 277-284
        • Starzl TE
        • Todo S
        • Fung J
        • Demetris AJ
        • Venkataramman R
        • Jain A
        FK506 for liver, kidney, and pancreas transplantation.
        Lancet. 1989; ii: 1000-1004
        • Tamura K
        • Woo J
        • Murase N
        • Carrieri G
        • Nalesnik MA
        • Thomson AW
        Suppression of autoimmune thyroid disease by FK 506: influence on thyroid infiltrating cells, adhesion molecule expression and anti-thyroglobulin antibody production.
        Clin Exp Immunol. 1993; 91: 368-375
        • Jegasothy BV
        • Ackerman CD
        • Todo S
        • Fung JJ
        • AbuElmagd K
        • Starzl TE
        Tacrolimus (FK506)—a new therapeutic agent for severe recalcitrant psoriasis.
        Arch Dermatol. 1992; 128: 781-785
        • Kino T
        • Hatanaka H
        • Miyata S
        • et al.
        FK506, a novel immunosuppressant isolated from a streptomyces II. Immunosuppressive effect of FK-506 in vitro.
        J Antibiot. 1987; 40: 1256-1265
        • Sawada S
        • Suzuki G
        • Kawase Y
        • Takaku F
        Novel immunosuppressive agent, FK506. In vitro effects on the cloned T cell activation.
        J Immunol. 1987; 139: 1797-1803
        • Bishop DK
        • Li W
        Cyclosporin A and FK506 mediate differential effects on T cell activation in vivo.
        J Immunol. 1992; 148: 1049-1054
        • Bruserud Ø
        • Pawelec G
        Cyclosporine A and FK506 show similar immunosuppressive effects on long-term in vitro T-cell proliferation.
        Int J Immunopharm. 1993; 15: 93-97
        • Schreiber SL
        • Crabtree GR
        The mechanism of action of cyclosporin A and FK506.
        Immunol Today. 1992; 13: 136-142
        • Schreiber SL
        Chemistry and biology of the immunophilins and their immunosuppressive ligands (Review).
        Science. 1991; 251: 283-287
        • Sikierka JJ
        • Sigal NH
        FK-506 and cyclosporin A: immunosuppressive mechanism of action and beyond.
        Curr Opin Immunol. 1992; 4: 548-552
        • Liu J
        • Farmer Jr, JD
        • Lane WS
        • Friedman J
        • Weissman I
        • Schreiber SL
        Calcineurin is a common target of cyclophilin-cyclosporin A and FKBP-FK506 complexes.
        Cell. 1991; 66: 807-815
        • Clipstone NA
        • Crabtree GR
        Identification of calcineurin as a key signaling enzyme in T-lymphocyte activation.
        Nature. 1992; 357: 695-697
        • McCaffrey PG
        • Perrino BA
        • Soderling TR
        • Rao A
        NF- ATp, a T lymphocyte DNA-binding protein that is a target for calcineurin and immunosuppressive drugs.
        J Biol Chem. 1993; 268: 3747-3752
        • de Paulis A
        • Stellato C
        • Cirillo R
        • Ciccarelli A
        • Oriente A
        • Marone G
        Anti-inflammatory effect of FK-506 on human skin mast cells.
        J Invest Dermatol. 1992; 99: 723-728
        • Hatfield SM
        • Mynderse JS
        • Roehm NW
        Rapamycin and FK506 differentially inhibit mast cell cytokine production and cytokine-induced proliferation and act as reciprocal antagonists.
        J Pharm Exp Therap. 1992; 261: 970-976
        • Dupuy P
        • Bagot M
        • Michel L
        • Descourt B
        • Dubertret L
        Cyclosporin A inhibits the antigen-presenting functions of freshly isolated human Langerhans cells in vitro.
        J Invest Dermatol. 1991; 96: 408-413
        • Kupper TS
        The activated keratinocyte: a model for inducible cytokine production by non-bone marrow-derived cells in cutaneous inflammatory and immune responses.
        J Invest Dermatol. 1990; 94: 146-150
        • Matsue H
        • Cruz Jr, PD
        • Bergstresser PR
        • Takashima A
        Cytokine expression by epidermal cell subpopulations.
        J Invest Dermatol. 1992; 99: 42-45
        • Buchsbaum ME
        • Kupper TS
        • Murphy GF
        Differential induction of intercellular adhesion molecule-1 in human skin by recombinant cytokines.
        J Cutan Pathol. 1993; 20: 21-27
        • Tokura Y
        • Yagi J
        • O'Malley M
        • et al.
        Superantigenic staphylococcal exotoxins induce T-cell proliferation in the presence of Langerhans cells or class II-bearing keratinocytes and stimulate keratinocytes to produce T- cell-activating cytokines.
        J Invest Dermatol. 1994; 102: 31-38
        • Strange P
        • Skov L
        • Baadsgaard O
        Interferon gammatreated keratinocytes activate T cells in the presence of superantigens: involvement of major histocompatibility complex class II molecules.
        J Invest Dermatol. 1994; 102: 150-154
        • Asami M
        • Kuno T
        • Mukai H
        • Tanaka C
        Detection of the FK506-FKBP-calcineurin complex by a simple binding assay.
        Biochem Biophys Res Commun. 1993; 192: 1388-1394
        • Gillitzer R
        • Berger R
        • Mielke V
        • Muller C
        • Wolff K
        • Stingl G
        Upper keratinocytes of psoriatic skin lesions express high levels of Math Eq mRNA in situ.
        J Invest Dermatol. 1991; 97: 73-79
        • Nickoloff BJ
        • Karabin GD
        • Barker JNWN
        • et al.
        Cellular localization of interleukin-8 and its inducer, tumor necrosis factor-alpha in psoriasis.
        Am J Pathol. 1991; 138: 129-140
        • Barker JNWN
        • Jones ML
        • Mitra RS
        • et al.
        Modulation of keratinocyte-derived interleukin-8 which is chemotactic for neutrophils and T lymphocytes.
        Am J Pathol. 1991; 139: 869-876
        • Tuschil A
        • Lam C
        • Haslberger A
        • Lindley I
        Interleukin-8 stimulates calcium transients and promotes epidermal cell proliferation.
        J Invest Dermatol. 1992; 99: 294-298
        • Larsen CG
        • Anderson AO
        • Oppenheim JJ
        • Matsushima K
        Production of interleukin-8 by human dermal fibroblasts and keratinocytes in response to interleukin-1 or tumour necrosis factor.
        Immunology. 1989; 68: 31-36
        • Barker JNWN
        • Sarma V
        • Mitra RS
        • Dixit VM
        • Nickoloff BJ
        Marked synergism between tumor necrosis factor-α and interferon-γ in regulation of keratinocyte derived adhesion molecules and chemotactic factors.
        J Clin Invest. 1990; 85: 605-608
        • Schulz BS
        • Michel G
        • Wagner S
        • et al.
        Increased expression of epidermal IL-8 receptor in psoriasis. Down-regulation by FK-506 in vitro.
        J Immunol. 1993; 151: 4399-4406
        • Furue M
        • Gaspari AA
        • Katz SI
        The effect of cyclosporin A on epidermal cells II. Cyclosporin A inhibits proliferation of normal and transformed keratinocytes.
        J Invest Dermatol. 1988; 90: 796-800
        • Ramirez-Bosca A
        • Kanitakis J
        • Haftek M
        • Faure M
        • Castells-Rodellas A
        • Thivolet J
        Effects of cyclosporin A on cultured human epidermal keratinocytes. Growth and 5-bromo-2′-deoxyuridine incorporation.
        Acta Derm Venereol. 1990; 70: 6-10
        • Dykes PJ
        • Brunt J
        • Marks R
        The effect of cyclosporin on human epidermal keratinocytes in vitro.
        Br J Dermatol. 1990; 122: 173-180
        • Sharpe GR
        • Fisher C
        Time-dependent inhibition of growth of human keratinocytes and fibroblasts by cyclosporin A: effect on keratinocytes at therapeutic blood levels.
        Br J Dermatol. 1990; 123: 207-213
        • Urabe A
        • Kanitakis J
        • Viac J
        • Thivolet J
        Cyclosporin A inhibits directly in vivo keratinocyte proliferation of living human skin.
        J Invest Dermatol. 1989; 92: 755-757
        • Ramirez-Bosca A
        • Kanitakis J
        • Thivolet J
        Cydosporin A exerts a cytostatic effect in vivo on human and murine epithelial cells.
        Cancer. 1990; 66: 936-940
        • Petzelbauer P
        • Wolff K
        Effects of cyclosporin A on resident and passenger immune cells of normal human skin and UV-induced erythema reactions.
        Br J Dermatol. 1992; 127: 560-565
        • Buttke TM
        • McCubrey JA
        • Owen TC
        Use of an aqueous soluble tetrazolium/formazan assay to measure viability and proliferation of tymphokine dependent cell lines.
        J Immunol Methods. 1993; 157: 233-240
        • Matsue H
        • Rothberg KG
        • Takashima A
        • Kamen BA
        • Anderson RGW
        • Lacey SW
        Folate receptor allows cells to grow in low concentrations of 5-methyltetrahydrofolate.
        in: 3rd ed. Proc Natl Acad Sci USA. 89. 1992: 6006-6009
        • Kondo S
        • Kono T
        • Sauder DN
        • McKenzie RC
        IL-8 gene expression and production in human keratinocytes and their modulation by UVB.
        J Invest Dermatol. 1993; 101: 690-694
        • Wong RL
        • Winslow CM
        • Cooper KD
        The mechanisms of action of cyclosporin A in the treatment of psoriasis.
        Immunol Today. 1993; 14: 69-74
        • Elder JT
        • Hammerberg C
        • Cooper KD
        • et al.
        Cyclosporin A rapidly inhibits epidermal cytokine expression in psoriasis lesions, but not cytokine-stimulated cultured keratinocytes.
        J Invest Dermatol. 1993; 101: 761-766
        • Kojima T
        • Cromie MA
        • Fisher OJ
        • Voorhees JJ
        • Elder JT
        GRO-α mRNA is selectively overexpressed in psoriatic epidermis and is reduced by cyclosporin A in vivo, but not in cultured keratinocytes.
        J Invest Dermatol. 1993; 101: 767-772
        • Shaw JP
        • Utz PJ
        • Durand DB
        • Toole JJ
        • Emmel EA
        • Crabtree OR
        Identification of a putative regulator of early T cell activation genes.
        Science. 1988; 241: 202-205
        • Stein B
        • Baldwin Jr, AS
        Distinct mechanisms for regulation of the interleukin-8 gene involve synergism and cooperativity between Math Eq and NF-κB.
        Mol Cell Biol. 1993; 13: 7191-7198
        • Lemster B
        • Rilo HR
        • Carroll PB
        • Nalesnik MA
        • Thomson AW
        FK506 inhibits cytokine gene and adhesion molecule expression in psoriatic skin lesions.
        Ann NY Acad Sci. 1993; 696: 250-256