Expression, localisation and functional activation of NFAT-2 in normal human skin, psoriasis, and cultured keratocytes.
Ciclosporin A (CsA) is widely utilized for the treatment of inflammatory skin diseases such as psoriasis. The therapeutic effects of CsA are thought to be mediated via its immunosuppressive action on infiltrating lymphocytes in skin lesions. CsA and tacrolimus block T cell activation by inhibiting the phosphatase calcineurin and preventing translocation from the cytoplasm to the nucleus of the transcription factor Nuclear Factor of Activated T cells (NFAT). As calcineurin and NFAT 1 have been shown to be functionally active in cultured human keratocytes, expression of other NFAT family members such as NFAT-2 and possible functional activation was investigated in human keratocytes. RT-PCR and Western Analysis were used to investigate the presence of NFAT-2 mRNA and protein in human keratocytes. Tissue culture of human keratocytes and immunostaining of cells on coverslips and confocal microscopy were used to assess the degree of nuclear localisation of NFAT-2 in cultured cells. Keratome biopsies were taken from patients with psoriasis (lesional and non-lesional skin) and normal skin and immunohistochemistry was used to assess the NFAT-2 localisation in these biopsies using a well characterized anti-NFAT-2 antibody. The NFAT-2 mRNA and protein expression was demonstrated using RT-PCR and Western blotting. Moreover, the expression of NFAT-2 in normal skin, non-lesional and lesional psoriasis showed a striking basal staining suggesting a role for NFAT-2 in keratocytes proliferation. A range of cell types in the skin express NFAT-2. The expression of NFAT-2 in human keratocytes and response to different agonists provides perhaps a unique opportunity to examine the regulation, subcellular localization and kinetics of translocation of different NFATs in primary cultured human cells. In these experiments the author assessed the expression, localization of NFAT-2 in cultured human keratocytes and measured the degree of nuclear localisaion of NFAT-2 using immunofluorescence and confocal microscopy and whether CsA and tacrolimus inhibit NFAT-2 nuclear translocation. As with NFAT 1, differentiation-promoting agents that increase intracellular calcium concentration induced nuclear translocation of NFAT-2 in cultured keratocytes but with different kinetics. These data provide the first evidence of that NFAT-2 is expressed in normal skin, psoriasis and that NFAT-2 functionally active in human keratocytes and that nuclear translocation of NFAT-2 in human skin cells has different kinetics than NFAT 1 suggesting that NFAT-2 may play an important role in regulation of keratocytes proliferation and differentiation at a different stage. Inhibition of this pathway in human epidermal keratocytes many account, in part for the therapeutic effects of CsA and tacrolimus in skin disorders such as psoriasis. Thus, supporting our previous work data that calcineurin/NFAT is functionally active not only in T cells, but in skin cells.
Al-Daraji
Wael I
WI
Malak
Tamer T
TT
Prescott
Richard J
RJ
Abdellaoui
Adel
A
Ali
Mahmud M
MM
Dabash
Tarek
T
Zelger
Bettina G
BG
Zelger
Bernhard
B
Mol Cell Biol. 1995 May;15(5):2697-706
7739550
Arch Dermatol Res. 1994;286(2):123-9
8154925
Cell Immunol. 1991 Apr 1;133(2):269-84
1707760
Dermatologica. 1990;181(2):117-21
2242778
J Clin Endocrinol Metab. 1989 Apr;68(4):851-4
2921314
Br J Dermatol. 1997 May;136(5):666-73
9205496
Immunity. 1995 May;2(5):461-72
7749981
Nature. 1997 Apr 24;386(6627):855-8
9126747
Blood. 1998 Apr 1;91(7):2223-30
9516119
J Biol Chem. 1996 Aug 23;271(34):20914-21
8702849
Immunol Today. 1994 Jun;15(6):274-81
8068174
J Immunol. 1997 Sep 15;159(6):2735-40
9300694
Cell Regul. 1990 Nov;1(12):965-73
1712634
Mol Cell Biol. 2000 Sep;20(17):6600-11
10938134
Nature. 1998 Mar 12;392(6672):182-6
9515963
Immunity. 1998 Jan;8(1):115-24
9462517
Mol Cell Biol. 1997 Apr;17(4):2066-75
9121455
Brain Res. 1987 May 12;411(1):120-8
2886179
Arch Dermatol Res. 1993;284(8):445-50
8466281
Nature. 1995 Apr 13;374(6523):617-22
7715701
Mol Biol Cell. 1998 Oct;9(10):2905-16
9763451
Proc Natl Acad Sci U S A. 2001 Aug 14;98(17):9575-80
11493684
Neurochem Int. 1995 Feb;26(2):135-43
7599534
J Dermatol Sci. 1994 Jul;7 Suppl:S47-54
7528050
J Immunol. 1990 Jan 1;144(1):251-8
1688572
Annu Rev Immunol. 1997;15:707-47
9143705
J Invest Dermatol. 1993 Dec;101(6):858-63
8245514
J Virol. 2001 Mar;75(6):3034-7
11222731
Mol Cell. 1998 Apr;1(5):627-37
9660947
Am J Pathol. 1998 Jul;153(1):39-45
9665463
J Biol Chem. 2000 May 12;275(19):14708-16
10799559
J Cell Sci. 1988 Aug;90 ( Pt 4):525-9
2473085
J Invest Dermatol. 1993 Jan;100(1):3-9
8423390
Cell. 1977 Jun;11(2):417-22
890737
Nature. 1993 Sep 23;365(6444):352-5
8397339
J Exp Med. 2000 Jan 3;191(1):9-22
10620601
J Exp Med. 1997 Jan 6;185(1):43-53
8996240
J Dermatol. 1991 May;18(5):252-7
1939851
J Exp Med. 1998 Jun 15;187(12):2031-6
9625762
J Cell Biol. 1978 Mar;76(3):705-11
24643
Br J Dermatol. 1995 Aug;133(2):157-67
7547380
Am J Pathol. 1995 Sep;147(3):718-27
7677183
Immunity. 1998 Nov;9(5):627-35
9846484
Nature. 1997 Apr 17;386(6626):728-31
9109491
Proc Natl Acad Sci U S A. 1998 Dec 22;95(26):15537-41
9861004
Nature. 1996 Oct 31;383(6603):837-40
8893011
J Exp Med. 1996 Jul 1;184(1):141-7
8691127
J Immunol. 1999 Jun 15;162(12):7294-301
10358178
Mol Cell Endocrinol. 2001 May 25;177(1-2):161-71
11377831
J Immunol. 1990 Feb 15;144(4):1418-24
1689353
Cancer Res. 1983 Sep;43(9):4443-7
6307514
Br J Dermatol. 1998 Jul;139(1):16-22
9764143
Immunity. 1998 Sep;9(3):295-304
9768749
J Invest Dermatol. 1992 Aug;99(2):193-200
1629632
Proc Natl Acad Sci U S A. 1992 Sep 1;89(17):8298-302
1381508
Immunity. 1997 Apr;6(4):361-9
9133415
J Immunol. 1989 Jul 15;143(2):718-26
2472451
Nature. 1992 Jun 25;357(6380):695-7
1377362
Immunity. 1998 Jan;8(1):125-34
9462518
J Biol Chem. 2000 Mar 31;275(13):9143-9
10734048
J Exp Med. 1996 Jul 1;184(1):101-12
8691123
Immunol Lett. 2001 Mar 1;76(2):133-8
11274732
Cell. 1999 Mar 5;96(5):611-4
10089876
Immunity. 2000 Apr;12(4):359-72
10795734
EMBO J. 1997 Aug 15;16(16):4983-98
9305640
Immunity. 1999 Feb;10(2):261-9
10072078
Front Biosci. 2000 Feb 01;5:D268-83
10704155
Cell. 1998 Apr 17;93(2):215-28
9568714
Science. 1996 Dec 13;274(5294):1903-5
8943202
Annu Rev Biochem. 1994;63:1045-83
7979236
Cancer Res. 1986 Apr;46(4 Pt 2):2068-71
2418960
Mol Cell Biol. 1996 Jul;16(7):3945-54
8668212
Pathobiology. 1992;60(2):93-9
1571097
EMBO J. 1996 Sep 2;15(17):4526-36
8887544
J Invest Dermatol. 1993 May;100(5):613-7
8098344
Immunity. 1997 Mar;6(3):235-44
9075924
Mol Cell Biol. 1997 May;17(5):2475-84
9111316
Nature. 1996 Jul 25;382(6589):370-3
8684469
Nature. 1998 Mar 12;392(6672):186-90
9515964
Int J Dermatol. 1985 Oct;24(8):507-8
2415474
J Virol. 2001 Oct;75(20):9955-65
11559828
EMBO J. 1990 Sep;9(9):2877-84
2118107
Clin Exp Dermatol. 2002 Oct;27(7):555-61
12464150
Nature. 1998 Mar 5;392(6671):42-8
9510247
Immunity. 1997 Dec;7(6):849-60
9430230
Immunity. 1999 May;10(5):515-24
10367897
Cell. 1979 Nov;18(3):681-94
42494
Biochem Soc Trans. 1997 May;25(2):347-54
9191115
J Invest Dermatol. 1993 Nov;101(5):732-7
8228336
Proc Natl Acad Sci U S A. 1996 Aug 20;93(17):8907-12
8799126
J Invest Dermatol. 2002 May;118(5):779-88
11982754
Science. 1996 May 10;272(5263):892-5
8629027
Curr Opin Cell Biol. 1993 Feb;5(1):70-6
8448032