Effects of dexamethasone and cyclosporin A on human beta-defensin in corneal epithelial cells.
Human beta-defensins (hBDs), which are mainly expressed in epithelial tissues, may contribute to infection-protective mechanisms in the ocular surface. We examined hBD1 and hBD2 expression in the ocular surface by RT-PCR and immunohistochemistry and studied the effects of immunosuppressive agents on their expression in human corneal epithelial cells in vitro. mRNA expression of hBD1 and hBD2 was confirmed in corneal epithelial cells. While the hBD1 peptide was immunohistochemically detected in corneal, limbal, and conjunctival epithelium, the level of expression was stronger in limbal- and conjunctival- than in corneal epithelium. Very weak expression of the hBD2 peptide was detected only in corneal epithelium. After 48-h culture of human corneal epithelial cells in the presence of 10(-5) M dexamethasone or 1 mg l(-1) cyclosporin A, total RNA was extracted and hBD1 and hBD2 mRNA expressions compared using semi-quantitative RT-PCR and introduced amplified fragment length polymorphism (iAFLP) assay. The two methods yielded almost identical results. hBD1 mRNA expression was not changed by dexamethasone but was down-regulated by cyclosporin A. hBD2 mRNA expression was up-regulated by dexamethasone and down-regulated by cyclosporin A. Our findings suggest that hBD1 and hBD2 are regulated by different mechanisms and that hBD1 may contribute to infection-protective mechanisms in the relatively immunosuppressed status induced by dexamethasone.
Department of Ophthalmology, Kyoto Prefectural University of Medicine, 465 Kajii-cho, Kamigyo-ku, Kyoto 602-084, Japan. tdk@seagreen.ocn.ne.jp
Kazuto
K
Terai
Yoichiro
Y
Sano
Satoshi
S
Kawasaki
Ken-ichi
K
Endo
Wakako
W
Adachi
Takeaki
T
Hiratsuka
Hirotoshi
H
Ihiboshi
Masamitsu
M
Nakazato
Shigeru
S
Kinoshita
D000328:Adult; D002478:Cells, Cultured; D016572:Cyclosporine / Q000494:pharmacology*; D003907:Dexamethasone / Q000494:pharmacology*; D019573:Epithelium, Corneal / Q000187:drug effects* / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D006801:Humans; D007166:Immunosuppressive Agents / Q000494:pharmacology*; D008297:Male; D011110:Polymorphism, Genetic; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D023083:beta-Defensins / Q000235:genetics / Q000378:metabolism*
C406407:DEFB1 protein, human; C106641:DEFB4A protein, human; D007166:Immunosuppressive Agents; D012333:RNA, Messenger; D023083:beta-Defensins; D003907:Dexamethasone; D016572:Cyclosporine