Therapeutic effects of tacrolimus ointment for refractory ocular surface inflammatory diseases.

PURPOSE: To investigate the therapeutic effects of topical tacrolimus ointment on refractory ocular surface inflammatory diseases. DESIGN: Retrospective interventional consecutive case series. PARTICIPANTS: Ten consecutive patients with severe ocular surface inflammatory diseases who were suspected to be steroid responders (elevation of intraocular pressure [IOP]) or were refractory to standard steroid therapy were studied. One patient had peripheral ulcerative keratitis with impending corneal perforation, 1 had a Mooren's ulcer, 2 had scleroperikeratitis, 5 had atopic keratoconjunctivitis, and 1 had vernal keratoconjunctivitis. METHODS: The clinical findings and therapeutic responses after treatment with 0.02% topical tacrolimus ointment were determined by conventional ophthalmological examinations. MAIN OUTCOME MEASURES: Resolution of the ocular surface diseases (e.g., decrease of hyperemia, ulceration, size of papillae) and IOP. The necessity to use steroids was also assessed. RESULTS: In all cases, marked to moderate improvement was obtained, including suppression of the melting reaction of the inflamed cornea, remission of scleroperikeratitis, and reduction of a giant papilla and corneal epithelial defect in severe atopic keratoconjunctivitis. The elevated IOP was reduced in steroid responders after successful cessation of steroid therapy. No adverse side effect was noted for 2 to 26 months of continuous treatment. CONCLUSIONS: Topical tacrolimus ointment is effective in treating refractory ocular surface inflammatory diseases and should be considered as an alternative to higher doses, steroid supplementation, or surgical intervention.

Clinical application of real-time polymerase chain reaction for diagnosis of herpetic diseases of the anterior segment of the eye.

PURPOSE: To assess the value of quantification of herpes simplex virus (HSV) DNA for the differential diagnosis of herpetic diseases of the anterior segment of the eye. METHODS: One hundred forty-four samples from 90 patients with ocular inflammatory diseases were examined for HSV DNA by real-time polymerase chain reaction (PCR) with primers set for the consensus sequence of HSV-1/2 DNA polymerase. The samples included corneal epithelial scrapings, tear fluid (200microl of eye wash), and aqueous humor. RESULTS: In cases of typical herpetic epithelial keratitis, a large number of copies of HSV DNA were detected (mean, 1.0 x 10(7) copies in epithelial scrapings and 3.5 x 10(5) copies in tear fluid). In atypical epithelial keratitis cases, a smaller number of HSV DNA copies were detected. In stromal keratitis cases, the number of copies of HSV DNA in the tear fluid (mean: 4.7 x 10(2) copies) was significantly smaller than in cases of epithelial keratitis. In the aqueous humor, the number of copies was small in endotheliitis cases (mean, 2.9 x 10(2) copies/microl), but the range was great, from (1.2-4.8) x 10(5)/microl in herpetic iridocyclitis. Seventeen percent of cases in which HSV was not suspected to be involved showed a small number of copies of HSV DNA, indicating the unexpected involvement of HSV in these cases. CONCLUSIONS: Real-time PCR is an informative method of diagnosing herpetic eye diseases and evaluating the possible involvement of HSV in other inflammatory ocular diseases.

Pathogenesis of herpetic stromal keratitis in CCR5- and/or CXCR3-deficient mice.

PURPOSE: Herpetic stromal keratitis (HSK) is an immunopathological reaction to herpes simplex virus type 1 (HSV-1) corneal infection. It has been reported that CD4+ cells play the most important role in the pathogenesis of this disease. In this study, we have focused on two chemokine receptors, CCR5 and CXCR3, which are expressed on CD4+ Th1 cells in mice HSK model. METHODS: CCR5-deficient (CCR5KO), CXCR3-deficient (CXCR3KO), CCR5/CXCR3 double-deficient (DKO), and wild type (WT) mice (C57/BL6 background) were infected intracorneally with HSV-1 (CHR3 strain). The corneas were examined biomicroscopically, and cryosections of the corneas were examined histologically and immunohistochemically. Real-time RT-PCR and RNase protection assay (RPA) were performed, and the virus titers were measured in excised eyes and trigeminal ganglia (TG). RESULTS: The HSK clinical severity in DKO mice was significantly lower than that in WT mice, and this was reversed by transfer of cells from the spleen of WT mice to DKO mice. Histologically, the numbers of T cells (CD4+ and CD8+ cells) and neutrophils infiltrating the cornea were significantly fewer in CCR5KO, CXCR3KO, and DKO mice. Transcript levels of immune-related cell surface marker in the eye by RPA were reduced in DKO mice. The expression of I-TAC was significantly increased in the cornea of CCR5KO mice, and MIP-1alpha and MIP-1beta were significantly lower in CXCR3KO mice than in WT mice by RT-PCR. There were no significant differences of virus titers in the eye and TG among any groups of mice except the increase in the TG of DKO mice on day 5 PI. CONCLUSIONS: The suppression of chemotaxis and activation of CD4+ Th1 cells by the lacking of CXCR3 and CCR5 causes a decrease of other infiltrating cells, resulting in a lower severity of HSK. These results suggest that targeting chemokine receptors is a promising way to treat HSK.