Koji Nishino

Intraocular pressure elevation after intravitreal or posterior sub-Tenon triamcinolone acetonide injection.

BACKGROUND Despite the benefits of intraocular steroids for the treatment of inflammatory, neovascular, proliferative, and edematous diseases, one of the side effects is raised intraocular pressure (IOP). In this study, we attempted to identify when IOP elevates, peaks, and returns to the preinjection baseline IOP after intravitreal or posterior sub-Tenon administration of triamcinolone acetonide, as well as the factors that might affect IOP. METHODS Retrospective case review was undertaken of 69 patients (82 eyes), who received either a 4 mg intravitreal (16 eyes) or a 20 mg posterior sub-Tenon (66 eyes) triamcinolone acetonide injection. IOP assessment for each eye was completed at the preinjection baseline and at the first, third, and sixth month of follow-up. RESULTS The mean IOP of all eyes increased significantly at each follow-up. The mean maximum elevation ratio from the baseline was 4.0 (SD 5.2) mm Hg. An elevation of 5 mm Hg or greater occurred in 28 eyes (34.1%). The maximum elevation correlated significantly with age (p < 0.01). The incidence of an elevation of 5 mm Hg or greater was significantly higher among patients younger than 60 years (p < 0.01) and relatively higher among female patients (p = 0.051). The mean IOP increased significantly at the first month after intravitreal injection but at all follow-up periods after posterior sub-Tenon injection. There was no significant difference in IOP elevation according to disease type, although eyes with diabetic retinopathy tended to be at higher risk of IOP elevation. Two eyes of two female patients, who had received posterior sub-Tenon injections for the treatment of diabetic retinopathy, required glaucoma surgery. INTERPRETATION The IOP elevation of 5 mm Hg or greater observed in 34.1% of the eyes was consistent with past reports. IOP elevation was associated with patients of less than 60 years of age and with female sex, and it lasted longer after posterior sub-Tenon injection than after intravitreal injection. Careful assessment of IOP during a follow-up period of at least 6 months is paramount, especially in younger female patients after posterior sub-Tenon injection.

Experimental immune-mediated blepharoconjunctivitis in rats induced by immunization with ragweed pollen

Abstract Background: A study was performed to compare the effects of immunization with ragweed pollen (RW) in two different adjuvants on the characteristics of a previously described model of experimental immune-mediated blepharoconjunctivitis (EC) in rats. Methods: Lewis or Brown Norway (BN) rats were immunized with 100 µg of RW in emulsion with aluminum hydroxide [Al(OH)3] or complete Freund’s adjuvant (CFA). Three weeks later, the animals were challenged with eye drops containing RW in PBS. Twenty-four hours after topical challenge, eyes, blood, and lymph nodes were obtained for histology, measurement of antigen-specific antibodies, and proliferation or cytokine assays, respectively. In addition to active immunization, recipients of RW-primed lymph node cells were challenged and evaluated as above. Results: RW in both adjuvants induced infiltration with predominantly mononuclear cells in Lewis rats and eosinophils in BN rats. As well as active immunization, eosinophils were detected only in BN rats by adoptive transfer of cells. Lymphocyte proliferative responses to RW were high in immunized Lewis rats when CFA was used as an adjuvant. In contrast, proliferative responses in BN rats were higher when Al(OH)3 was used. RW-specific IgE was detected only in BN rats. There were no significant differences in RW-specific IgG1/ IgG2a ratio among the four groups. Lewis rats had higher level of RW-specific interferon-γ in the culture supernatant. Conclusions: The characteristics of EC are different in Lewis and BN rats, dependent on the genetic background of the rat strains. The response to RW was similar to other previously used antigens, such as ovalbumin.

Suppression of experimental immune-mediated blepharoconjunctivitis in Brown Norway rats by topical application of FK506

Abstract.Background: Experimental immune-mediated blepharoconjunctivitis (EC) in Brown Norway (BN) rats, which is inducible by transfer of antigen-specific T cells, is a model for human allergic conjunctivitis. We investigated the possible inhibition of EC in BN rats by topical application of FK506, which is an immunosuppressive agent that mainly targets T cells. Methods: To induce EC by active immunization, ovalbumin (OVA) adsorbed to alum was injected into the hind footpads of BN rats. Three weeks after the initial immunization, rats were challenged with OVA by eye drops. Twenty-four hours later, lids including conjunctivas, lymph nodes (LNs), and sera were harvested for histology or reverse transcriptase PCR, proliferation assays, and measurement of IgE titer, respectively. For passive immunization, rats were intravenously injected with 10 million of in vitro-stimulated OVA-primed LN cells. Four days after the transfer, rats were challenged with OVA and evaluated as above. The rats were divided into two groups. One group received topical FK506 treatment three times per day from 15 to 21 days after active immunization or from 1 to 4 days after transfer. The other group was treated with vehicle as above. Results: FK506 treatment suppressed infiltration of both lymphocytes and eosinophils in the conjunctiva either by active or passive immunization (P<0.002). No differences were noted in antigen-specific cellular and humoral immune responses. Concerning cytokine expression in the conjunctiva, a prominent difference was noted only with IL-4, which was more abundantly detected in the vehicle-treated group. Conclusion: Topical FK506 treatment suppressed EC in BN rats, possibly by inhibition of IL-4 in the conjunctiva.

Analysis of effects of stimulation in vitro of ovalbumin primed lymph node cells on adoptive transfer of experimental immune mediated blepharoconjunctivitis in Lewis rats

AIM To analyse the role of stimulation in vitro of lymphocytes on the augmentation of experimental immune mediated blepharoconjunctivitis (EC, formerly EAC) in Lewis rats induced by adoptive transfer. METHODS Two weeks after immunisation with ovalbumin (OVA), rat draining lymph nodes were collected and 50 × 106 cells were injected into naive syngeneic recipients either directly or after culture in vitro with OVA, concanavalin A (Con A), or purified protein derivative (PPD) for 3 days. Four days after injection the rats were topically challenged with OVA. 24 hours later, they were sacrificed and eyes and spleens were harvested for histology and proliferation assay. In some experiments, naive recipient rats were irradiated with 7 Gy γ ray before transfer. The expression of adhesion molecules and cytokine profile of OVA primed lymph node cells were also investigated. RESULTS Both infiltrated cell number and splenocyte proliferation in the recipients of stimulated cells were higher than those of unstimulated cells. In vitro stimulation with OVA or Con A induced a severe cellular infiltration, while stimulation with PPD did not. Irradiation markedly diminished cellular infiltration. Stimulation in vitro upregulated the CD4/CD8 ratio by four times and augmented expression of CD25, I-A, ICAM-1 molecules on OVA primed lymph node cells by about five times. IFN-γ was detected in OVA primed cells by stimulation in vitro, while IL-4 mRNA was extinguished by stimulation in vitro. CONCLUSIONS Augmentation of EC by stimulation in vitro of transferred lymphocytes might depend on the upregulation of expression of cell surface molecules and cytokine shift as well as augmented antigen specificity.

Characterization of the immunopathogenic responses to ovalbumin peptide 323–339 in experimental immune-mediated blepharoconjunctivitis in Lewis rats

PURPOSE We recently reported the essential role of cellular immunity on the induction of experimental immune-mediated blepharoconjunctivitis (EC, formerly EAC) by using ovalbumin (OVA) as a model antigen in Lewis rats. The purpose of this study was to investigate the possible induction of EC by immunization with an OVA peptide (OVA 323-339). METHODS Lewis rats were immunized with various doses of OVA or OVA 323-339 in complete Freunds adjuvant. Three weeks later they were challenged with OVA or OVA 323-339 by eye drops; 24 h after challenge, eyes including lids, lymph nodes and blood were harvested after clinical evaluation. An OVA 323-339-specific cell line (S816) was established by periodical stimulation with this peptide. Pathogenicity of S816 was tested by adoptive transfer of S816 into syngeneic recipient rats after challenge with OVA or OVA 323-339. RESULTS All rats immunized with OVA 323-339 developed EC after challenge with OVA or OVA 323-339. Rats immunized with OVA 323-339 at doses as low as 0.01 microg had severe clinical scores. OVA-primed rats also developed EC after challenge with OVA 323-339. OVA-primed lymph node cells responded to OVA but not to OVA 323-339. OVA 323-339-primed lymph node cells responded to OVA 323-339 but not to OVA and produced IFN-gamma by stimulation with either OVA or OVA 323-339 (three- to fourfold more than with OVA-primed lymph node cells). Recipient rats of S816 developed severe EC after challenge with either OVA or OVA 323-339. CONCLUSION OVA 323-339 was identified as a potent pathogenic peptide in EC.

Inflammatory reaction following cataract surgery and implantation of acrylic intraocular lens in rabbits with endotoxin-induced uveitis.

PurposeTo investigate whether inflammatory responses are more severe in uveitic eyes than nonuveitic eyes when acrylic intraocular lens (IOL) is implanted after cataract surgery.MethodsClear lens removal (phacoemulsification and aspiration) was conducted and the hydrophobic acrylic IOL (AR40e, AMO) was implanted in adult albino rabbits. Just after the operation, rabbits were divided into two groups. One group (nine rabbits) received intravitreal injection of lipopolysaccharide (LPS, 200 ng/10 μl) into both eyes to induce endotoxin-induced uveitis (EIU) and the other group (nine rabbits) received intravitreal injection of phosphate-buffered saline (PBS, 10 μl) into both eyes as the control. Aqueous humour (AH) and IOLs were harvested 1, 3 , and 7 days after the intravitreal injection. The infiltrating cell number in AH was counted and the protein concentration of AH was measured. IOLs were evaluated morphologically.ResultsAt 1 day after intravitreal injection, both the infiltrating cell number in AH and protein concentration of AH were significantly higher in the LPS-injected group than in the PBS-injected group. Similarly, more inflammatory cells attached to the surfaces of the IOLs in the LPS-injected group. However, 7 days later, inflammatory reactions subsided and no clear differences in any of the parameters examined were observed between the two groups.ConclusionsAt 7 days after the operation, inflammatory reactions in eyes implanted with the hydrophobic acrylic IOLs were similar in uveitic eyes and nonuveitic eyes. The data suggest that the hydrophobic acrylic IOLs may be suitable for patients with uveitis.

Treatment with FTY720 during the induction or effector phase suppresses the development of experimental allergic conjunctivitis in mice

Purpose: To investigate whether experimental allergic conjunctivitis (EC) can be suppressed by treatment with the immunomodulatory drug FTY720, which reduces the recruitment of effector T cells into inflammatory sites.

Vogt-Koyanagi-Harada Disease with Onset in Elderly Patients Aged 68 to 89 Years

BackgroundWe present the clinical case reports of elderly Vogt-Koyanagi-Harada (VKH) disease patients, including the oldest patient at onset to date.CasesFour patients with VKH disease, whose ages at onset were 68 to 89 years, were treated at Kochi Medical School Hospital between December 2002 and September 2004.ObservationsFour elderly patients were diagnosed with VKH disease according to the International Revised Diagnostic Criteria for VKH disease. Two were treated with corticosteroid pulse therapy, but the other two could not tolerate high-dose steroid therapy and were treated with only topical corticosteroids. All had recurrences, which were treated effectively. None of these patients experienced recurrence after sunset glow fundus was observed in the follow-up period.ConclusionsVKH disease is thought to be more common in younger people, but it is not as uncommon in elderly people as is generally believed. Among elderly VKH disease patients, some cannot tolerate high-dose corticosteroid therapy, so we need to treat these patients with only topical corticosteroids. Jpn J Ophthalmol 2007;51:60–63

Analysis of Responses of Peripheral Blood Lymphocytes From Sarcoidosis Patients to Purified Protein Derivative

PURPOSE Negative skin test reaction against purified protein derivative (PPD) is one of the important diagnostic criteria for sarcoidosis. The purpose of this study was to investigate the relationship between negative skin test reaction against PPD and the responses of peripheral blood lymphocytes (PBLs) against PPD in sarcoidosis patients. METHODS Sarcoidosis patients (n = 14) with ocular changes were selected for this study. As a control, blood was collected from volunteers without uveitis (n = 10). All subjects in both groups had a history of bacille de Calmette-Guerin vaccination, and 8 of the 14 patients underwent the PPD skin test. Peripheral blood lymphocytes were obtained from all patients and separated by gradient centrifugation. Peripheral blood lymphocytes were tested for their proliferative responses to PPD. To determine the frequency of PPD-specific precursor cells in blood, limiting dilution analysis was used. The frequency was calculated following the Poisson model. The culture supernatants were harvested 3 days after incubation and assayed for interferon (IFN)-gamma production by ELISA. RESULTS Four of the 8 patients showed a negative skin test reaction against PPD, while all the healthy volunteers had a positive reaction. Compared to PBLs from healthy volunteers, PBLs from sarcoidosis patients showed weaker responses to PPD. There was no clear difference between these two groups about frequency of precursor cells against PPD and production of IFN-gamma. CONCLUSIONS Negative skin test for PPD in sarcoidosis patients does not seem to have any relationship to the low precursor frequency of PPD-specific cells in patient blood.

Suppression of induction of experimental immune mediated blepharoconjunctivitis by tolerogenic conjugates of the antigen and monomethoxypolyethylene glycol

AIM Covalent conjugates consisting of diverse antigens coupled to optimal numbers of monomethoxypolyethylene glycol (mPEG) molecules have been shown to suppress antigen specific antibody formation. In this study, the possibility was examined that the same conjugates might prevent experimental immune mediated blepharoconjunctivitis (EC, formerly EAC) which had been shown to be caused by CD4+ T cells—that is, to cell mediated immunity. METHODS 6–8 week old male Lewis rats were used. The test groups of rats received two intravenous injections, each of 300 μg, of a conjugate of ovalbumin mPEG (OVA(mPEG)11) in phosphate buffered saline (PBS), 14 and 28 days before the single immunisation with OVA in complete Freund’s adjuvant. The rats were challenged 3 weeks later by eye drops containing OVA; 24 hours later they were sacrificed, and their eyes, blood, and lymph nodes were harvested for histological examination and determination of anti-OVA antibody titres and levels of cellular immunity. Two control groups received PBS or OVA in PBS before immunisation. Furthermore, the possibility that OVA(mPEG)11may have induced OVA specific suppressor cells was tested by establishing the effects of the co-transfer of splenocytes from OVA(mPEG)11 treated rats with OVA primed lymph node cells on the manifestations of EC. RESULTS Either PBS or OVA pretreated rats, which had not received OVA(mPEG)11, developed high levels of antibodies and cell mediated immune responses to OVA, and application of eye drops led to blepharoconjunctivitis with massive cellular infiltration. In contrast, pretreatment with OVA(mPEG)11 prevented cellular infiltration into the lids and conjunctivas, as well as the formation of detectable humoral and cellular immunity against OVA. Co-transfer of splenocytes from OVA(mPEG)11 treated rats with OVA primed lymph node cells suppressed the cellular infiltration on application of OVA on the conjunctiva. CONCLUSIONS These data indicate that intravenous injection of OVA(mPEG)11conjugates suppressed both humoral and cellular immunity by the effects of antigen specific suppressor cells, thus leading to the inhibition of development of EC.