Anti-CD40 monoclonal antibody ameliorates experimental autoimmune uveoretinitis in mice.

OBJECTIVE: To investigate the effects of CD40 on ocular inflammation in experimental autoimmune uveoretinitis (EAU) in B10.RIII mice. ANIMALS STUDIED: EAU-susceptible B10.RIII mice were subcutaneously immunized with interphotoreceptor retinoid-binding protein (IRBP) 161-180 in complete Freund's adjuvant and evaluated clinically and pathologically on days 7, 14, 21, 28, and 35 postimmunization. Anti-CD40 antibody was intraperitoneally injected into mice every other day from days 7 to 14 postimmunization. Phosphate-buffered saline (PBS)-injected EAU mice were used as the controls. PROCEDURES: The frequencies of CD11c+ CD40+ dendritic cells (DCs), CD11c+ MHC-II+ DCs, and CD11c+ CD40+ MHC-II+ DCs in splenocytes were evaluated by flow cytometry on days 0, 7, 14, and 21 after immunization. Tumor necrosis factor (TNF)-α and interleukin (IL)-6 production in CD11c+ DCs was assessed by ELISA. IRBP-specific lymphocyte proliferation was assessed using a modified MTT cell proliferation assay. RESULTS: The number of CD11c+ CD40+ DCs, CD11c+ MHC-II+ DCs, and CD11c+ CD40+ MHC-II+ DCs increased at the onset of EAU, peaked at the height of disease severity, and was sustained at a high level until day 21. Treatment with anti-CD40 antibody significantly alleviated clinical and pathological activities related to EAU. Compared with the control mice, antibody-treated EAU mice showed few CD11c+ CD40+ DC and CD11c+ CD40+ MHC-II+ DC frequencies in splenocytes. The anti-CD40 antibody significantly suppressed IRBP-specific lymphocyte proliferation and TNF-α and IL-6 production by DCs in EAU mice. CONCLUSIONS: The increased expression of CD40 and major histocompatibility complex (MHC) class II molecules in the splenocytes of EAU mice were correlated with inflammatory activity. Anti-CD40 treatment can significantly attenuate EAU activity by inhibiting systemic IRBP-specific immune responses.

Laser-induced intrachoroidal dexamethasone drug delivery system to posterior eye segment.

PURPOSE: To investigate the feasibility of laser-induced intrachoroidal dexamethasone (DEX) delivery as a potentially useful therapy for adjusting the most effective drug level to the posterior segment eye diseases. METHODS: An implant was prepared by dissolving poly(DL-lactide) and DEX. In vitro release of DEX was evaluated at 7, 14, and 28 days by ELISA. In vivo, a DEX implant was inserted into a rabbit choroid, and 10, 50, or 200 burns of photocoagulation were applied at the implant lesion. After treatment, the vitreous humor was immediately aspirated and the DEX level was measured by liquid chromatography/mass spectrometry/mass spectrometry. Furthermore, the vitreous DEX level was measured at 1, 7, 14, and 28 days after implantation and 50 burns of photocoagulation. The toxicity of the laser-induced DEX implant was evaluated by ophthalmoscopy and light microscopy. Endotoxin-induced uveitis (EIU) was induced after DEX implantation and photocoagulation, and anti-inflammatory activities were evaluated by grading clinical signs, protein concentrations, and histopathologic studies. RESULTS: Photocoagulation significantly increased the DEX release from the implant at 7 days in vitro. In vivo, the DEX implant exposed to 10, 50, and 200 burns of photocoagulation increased the vitreous DEX levels in a dose-dependent manner. The vitreous DEX level in the DEX implant applied to 50 burns of photocoagulation peaked 1 day after treatment. The laser-induced DEX implant showed no retinal abnormalities except the implantation site, and significantly inhibited the EIU. CONCLUSIONS: Laser-induced intrachoroidal DEX delivery controls the DEX level in the vitreous humor and effectively prevents the experimental uveitis.

Effects of systemic and intravitreal TNF-α inhibition in experimental autoimmune uveoretinitis.

PURPOSE: To investigate the effect of systemic or local TNF-α inhibition with etanercept on experimental autoimmune uveoretinitis (EAU). METHODS: EAU was induced by immunizing B10.RIII mice with IRBPp161-180 or by adoptively transferring uveitogenic splenocytes. Mice received systemic or local treatment with etanercept in the afferent or efferent phase. For systemic treatment, mice were injected intraperitoneally. For local treatment, etanercept was injected intravitreally or subconjunctivally. Control mice received PBS. EAU scores were determined histologically. Splenic cells were assessed for [(3)H]thymidine incorporation. ELISA was performed to measure levels of cytokines produced by splenocytes. Vitreous cavity-associated immune deviation (VCAID) was induced by intravitreally injecting ovalbumin and evaluated by measuring DTH reaction. RESULTS: After systemic treatment with etanercept in the afferent phase, EAU disease scores, IRBP-specific cell proliferation, and production of Th1, Th2, and Th17 cytokines were reduced. EAU also improved after intravitreal etanercept treatment in the afferent phase, with unaltered IRBP-specific proliferation, reduced IFN-γ, but increased IL-6 and IL-10 secretion. VCAID induction was impaired after intravitreal etanercept treatment. No amelioration of EAU or reduction in IRBP-specific cell response was found after systemic or intravitreal treatment in the efferent phase or after subconjunctival treatment. After adoptive transfer, etanercept- and PBS-treated recipients showed similar disease severity and antigen-specific proliferation of splenocytes. CONCLUSIONS: It can be concluded that TNF-α participates mainly in the immunopathology in the induction phase of EAU. The mechanism of action underlying EAU improvement may be different for local and systemic etanercept treatment.

Everolimus improves experimental autoimmune uveoretinitis.

The efficacy and action mechanism of everolimus in the treatment of experimental autoimmune uveoretinitis (EAU) was analyzed. Disease was induced in B10.RIII mice by immunization with human interphotoreceptor-retinoid-binding protein peptide 161-180 (hIRBPp161-180). Everolimus was administered by oral gavage (5 mg/kg/d) beginning either two days before or 14 days after immunization. Everolimus significantly reduced the histopathological uveitis score compared to sham-treated mice. To examine the effect on the antigen-specific immune response, proliferation ([(3)H]-thymidine test) and delayed-type hypersensitivity (DTH) response were measured. Furthermore, content of T-helper-1, -2, and -17 cytokines were analyzed intraocularly (Bead Array) and in cell culture supernatants from splenocytes (sandwich ELISA). To study the effect on the humoral immune response the presence of antigen-specific serum antibodies was tested (indirect ELISA). The DTH, the humoral immune response, the proliferation of splenocytes and the intraocular Th1, Th2, Th17 cytokine content and in vitro production of Th1 and Th17 cytokines were impaired after everolimus treatment. The study of CD4+CD25+FoxP3+ regulatory T cells (T(reg)) in peripheral blood, draining lymph nodes, and spleen by flow cytometry showed an increased number of splenic T(reg) in mice of the everolimus therapy group. Furthermore the T(reg) of these mice had a higher suppressive capacity than cells from sham-treated mice. These results indicate that the immunosuppressive effect of everolimus on EAU was associated with the suppression of pathogenic effector responses and induction of regulatory T cells.

A promising therapeutic approach for treatment of posterior uveitis: recombinant T cell receptor ligand protects Lewis rats from acute and recurrent experimental autoimmune uveitis.

INTRODUCTION: Chronic autoimmune uveitis is a major cause of vision loss from intraocular inflammation in humans. In this study we report that a recombinant TCR ligand (RTL220) composed of the alpha1 and beta1 domains of MHC class II molecules linked to the uveitogenic interphotoreceptor retinoid-binding protein (IRBP) 1177-1191 peptide is effective in the suppression of acute and recurrent experimental autoimmune uveitis (EAU). MATERIAL AND METHODS: EAU was induced with IRBP1177-1191 peptide or by adoptive transfer of specific T cells in Lewis rats. The rats received 5 doses of RTL220 subcutaneously every other day starting at the onset of clinic signs of EAU. RESULTS: The administration of RTL220 resulted in a delayed onset and a significant amelioration of the disease severity at clinical levels and showed protection of the retina from inflammatory damage at histological levels. In treatment of recurrent EAU, RTL220 administrated at the first or second onset of clinical disease significantly inhibited EAU, modulated immune responses and provided protection from relapses of uveitis. The systemic and local proinflammatory cytokines were significantly reduced, including IL-17. There was local and systemic increase in IL-10 and reduction in the expression of the proinflammatory chemokines CCL2, CCL3 and CCL5. CONCLUSIONS: Our studies demonstrate a successful treatment of acute and recurrent EAU with RTL220, which effectively suppressed the recurrence of inflammation and reversed clinical and histological EAU by altering cytokine and chemokine expression. These findings strongly support a possible clinical application of this novel class of peptide/MHC class II drugs for patients with autoimmune uveitis.

Evaluation of experimental autoimmune uveitis in mice treated with FTY720.

PURPOSE: FTY720 (fingolimod) is an immunomodulatory drug capable of preventing T-cell migration to inflammatory sites by binding to and subsequently downregulating the expression of sphingosine-1 phosphate receptor 1 (S1P(1)) leading in turn to T-cell retention in lymphoid organs. Additional effects of FTY720 by increasing functional activity of regulatory T cells have recently been demonstrated, raising the conversion of conventional T cells into regulatory T cells and affecting the sequestration of regulatory T cells in normal mice. In this study, the action of FTY720 in the ocular autoimmune model in mice was investigated. METHODS: Mice were immunized with 161-180 peptide and pertussis toxin and were treated with 1 mg/kg/d FTY720 by gavage (7-21 days postimmunization [dpi]) or left untreated. Spleen cells, harvested 21 dpi, were cultured and assayed for cytokine production. Draining lymph node, spleen, and eye cells 21 dpi were assayed for quantification of T-cell populations. Disease severity was evaluated by histologic examination of the enucleated eyes at 21 and 49 dpi. In addition, anti-IRBP antibodies were analyzed by ELISA. RESULTS: FTY720 was effective in suppressing the experimental autoimmune uveitis score. Although there was a reduction in the number of eye-infiltrating cells, FTY did not prevent Treg accumulation at this site. FTY720 leads to a significant increase of CD4(+)IFN-gamma(+) and CD4(+)Foxp3(+) cell percentages in lymph nodes, suggesting that this site could be the source of Treg cells found in the eye. CONCLUSIONS: The data showed that treatment in vivo with FTY720 was able to suppress EAU in mice. These results are indicative of the possible therapeutic use of FTY720 in ocular autoimmune processes.

Toxoplasmosis ocular / Ocular toxoplasmosis

La toxoplasmosis ocular es una enfermedad producida por el parásito toxoplasma gondii. Es la causa más frecuente de uveítis posterior, es una enfermedad de distribución universal, al menos 500 millones de personas están infectadas en todo el mundo, ocasionando disminución de la visión y ceguera en muchas de ellas. Por tal motivo, realizamos una revisión actualizada sobre, la situación actual a nivel mundial, la historia de la enfermedad, la prevención, formas clínicas y el control de la toxoplasmosis. Se tratan otros aspectos de interés como el modo de transmisión, los hospederos (definitivos e intermediarios) y las manifestaciones clínicas más notables

The ocular toxoplasmosis is an illness caused by Toxoplasma gondii parasite. It is the most frequent cause in posterior uveitis, and it spreads worldwide since at least 500 million people are infected in the entire world, causing decrease of vision and blindness in many of them. This is the reason why we made a literature review, the current situation worldwide, the history, the prevention, the clinical forms and the control of toxoplasmosis. Other interesting aspects were the channel of transmission, the hosts (intermediary and final) and the most remarkable clinical manifestations

Toxoplasmosis ocular / Ocular toxoplasmosis

La toxoplasmosis ocular es una enfermedad producida por el parásito toxoplasma gondii. Es la causa más frecuente de uveítis posterior, es una enfermedad de distribución universal, al menos 500 millones de personas están infectadas en todo el mundo, ocasionando disminución de la visión y ceguera en muchas de ellas. Por tal motivo, realizamos una revisión actualizada sobre, la situación actual a nivel mundial, la historia de la enfermedad, la prevención, formas clínicas y el control de la toxoplasmosis. Se tratan otros aspectos de interés como el modo de transmisión, los hospederos (definitivos e intermediarios) y las manifestaciones clínicas más notables(AU)

The ocular toxoplasmosis is an illness caused by Toxoplasma gondii parasite. It is the most frequent cause in posterior uveitis, and it spreads worldwide since at least 500 million people are infected in the entire world, causing decrease of vision and blindness in many of them. This is the reason why we made a literature review, the current situation worldwide, the history, the prevention, the clinical forms and the control of toxoplasmosis. Other interesting aspects were the channel of transmission, the hosts (intermediary and final) and the most remarkable clinical manifestations(AU)

Role of DAF in protecting against T-cell autoreactivity that leads to experimental autoimmune uveitis.

PURPOSE: To investigate the role of decay-accelerating factor (DAF), a cell surface complement regulator that recently has been linked to T-cell responses and autoimmunity in the pathogenesis of experimental autoimmune uveitis (EAU). METHODS: EAU was induced in wild-type (WT) and Daf1(-/-) mice, and their disease severities, IRBP specific Th1/Th17 responses, and cytokine expression profiles were compared. In a test of the efficacy of treatment with soluble mouse DAF protein, EAU was induced in disease-susceptible B10.RIII mice, and they were treated with 0.5 mg soluble DAF protein or equal volume of PBS IP every other day. Retinal histology and IRBP-specific T-cell responses were compared after 14 days. RESULTS: Both EAU incidence and histopathology scores were significantly greater in Daf1(-/-) mice. There was a >10-fold greater mononuclear cell influx into the retina together with severe vasculitic lesions, retinal folding, and photoreceptor cell layer destruction. There were 5- to 7-fold greater Th1 and 3- to 4-fold greater Th17 responses against IRBP in Daf1(-/-) mice with EAU, and they expressed significantly elevated levels of GM-CSF, IL-2, IL-3, and IFN-gamma. WT B10.RIII mice that received soluble DAF protein treatments exhibited decreased IRBP-specific Th1/Th17 responses and were protected from retinal injury compared with the mice that received PBS treatments. CONCLUSIONS: DAF significantly influences IRBP-specific Th1 and Th17 responses and disease severity in EAU. Systemic upregulation of DAF levels could be used to suppress retinal antigen(s)-specific autoimmunity to treat autoimmune posterior uveitis.

Neuroprotective effect of an antioxidant, lutein, during retinal inflammation.

PURPOSE: Lutein has been the focus of recent study as a possible therapeutic approach for retinal diseases, but the molecular mechanism of its neuroprotective effect remains to be elucidated. The aim of this study was to investigate, with the use of a mouse endotoxin-induced uveitis (EIU) model, the neuroprotective effects of lutein against retinal neural damage caused by inflammation. METHODS: EIU was induced by intraperitoneal injection of lipopolysaccharide (LPS). Each animal was given a subcutaneous injection of lutein or vehicle three times: concurrently with and 3 hours before and after the LPS injection. Analysis was carried out 24 hours after EIU induction. Levels of rhodopsin protein and STAT3 activation were analyzed by immunoblotting. Lengths of the outer segments of the photoreceptor cells were measured. Dark-adapted full-field electroretinograms were recorded. Oxidative stress in the retina was analyzed by dihydroethidium and fluorescent probe. Expression of glial fibrillary acidic protein (GFAP) was shown immunohistochemically. RESULTS: The EIU-induced decrease in rhodopsin expression followed by shortening of the outer segments and reduction in a-wave amplitude were prevented by lutein treatment. Levels of STAT3 activation, downstream of inflammatory cytokine signals, and reactive oxygen species (ROS), which are both upregulated during EIU, were reduced by lutein. Pathologic change of Müller glial cells, represented by GFAP expression, was also prevented by lutein. CONCLUSIONS: The present data revealed that the antioxidant lutein was neuroprotective during EIU, suggesting a potential approach for suppressing retinal neural damage during inflammation.

Administration of a peptide inhibitor of alpha4-integrin inhibits the development of experimental autoimmune uveitis.

PURPOSE: Recruitment of lymphocytes into the retina and to the vitreous during the development of experimental autoimmune uveitis (EAU) is governed by factors such as the state of activation of inflammatory cells and the repertoire of adhesion molecules expressed by the local vascular endothelia. alpha4 Integrins and their receptors play an important role during homing of cells to the inflammatory site. In the present study, the effect of alpha4-integrin inhibitor on the development of EAU was investigated. METHODS: EAU was induced either by immunizing B10.RIII mice with the 161-180 peptide or by adoptive transfer of interphotoreceptor retinoid-binding protein (IRBP)-specific uveitogenic T cells. Animals were treated with an active peptide inhibitor (alpha4-api) or a peptide control at different time points after induction of disease. EAU was evaluated by histology 21 to 49 days after immunization. Antigen-specific cell proliferation was evaluated by thymidine incorporation. Cytokine synthesis in culture supernatants and anti-IRBP-specific serum IgG1 and IgG2a were evaluated by ELISA. Delayed-type hypersensitivity was evaluated by ear challenge 2 days before the termination of the experiment. RESULTS: Treatment with alpha4-api had a significant ameliorating effect on EAU. The anti-IRBP antibody response and cellular proliferation were not affected by the treatment, whereas delayed-type hypersensitivity was significantly diminished. Cytokine synthesis was not changed by treatment, except for a decrease in IL-10 levels. CONCLUSIONS: The results show that small-molecule inhibitors of alpha4-integrins can act therapeutically in EAU, possibly by interfering with cell adhesion events involved in the development of the disease.

Anti-CD137 mAb treatment inhibits experimental autoimmune uveitis by limiting expansion and increasing apoptotic death of uveitogenic T cells.

PURPOSE: To explore the role of CD137 in the pathogenesis of experimental autoimmune uveitis (EAU) and to compare the inhibitory mechanism of anti-CD137 mAb with other costimulatory blockers. METHODS: EAU was induced in B10RIII mice, either by immunization with a uveitogenic peptide, IRBP161-180, derived from the interphotoreceptor retinoid-binding protein, or by adoptive transfer of IRBP161-180-specific T cells. The effect of an agonistic anti-CD137 mAb (2A) on the in vivo induction of disease was studied. Subsequently, the mechanism by which anti-CD137 mAb inhibits uveitogenic T-cell activation was investigated, by using the adoptive transfer of T cells derived from anti-CD137 mAb-treated mice, and in vitro, using the proliferative response and apoptotic cell death of IRBP-specific T cells from anti-CD137 mAb-treated mice. RESULTS: Administration of anti-CD137 mAb prevented the development of de novo induced uveitis, but not that induced by adoptive transfer of pathogenic T cells. Furthermore, anti-CD137 mAb treatment of the animals resulted in decreased expansion of uveitogenic T cells, accompanied by increased activated cell death and resistance to reinduction of uveitis. CONCLUSIONS: CD137 plays a critical role in the induction, rather than the effector, phase of the disease. Different costimulatory molecules have different effects on the activation of autoreactive T cells by acting in different phases of T-cell activation.

Anti-inflammation and antifibroblastic actions of interleukin-1 blockers.

Interleukin-1 (IL-1) blockers, CK 127 and CK 129, were found to inhibit IL-1-induced posterior uveitis very effectively at 3-10 mg/kg i.p. and were more potent than prednisolone which required at least 20 mg/kg i.p. to achieve the same level of anti-uveitis action. CK 127 and CK 129 were also found to be effective in inhibiting fibroblast-like corneal cells at 30-300 micrograms/ml and conjunctival cells at 0.3-10 micrograms/ml. These results indicate that IL-1 blockers are more potent in inhibiting the cell growth of conjunctival cells than that of corneal cells. From in vitro cell culture experiments, it was found that inhibition of cell growth could be due primarily to the inhibition of DNA. Although the inhibition of cell growth was due mainly to the inhibition of DNA synthesis, mRNA synthesis was also markedly inhibited. In both cells, the protein synthesis was unaffected in a few cases and markedly stimulated in most cases.

Antiuveitis and inhibition of fibroblast-like corneal and conjunctival cell cultures by interleukin-1 blockers, CK 125, CK 126 and CK 128.

Three new interleukin-1 (IL-1) blockers, CK 125, CK 126 and CK 128, were studied for their effects on IL-1-induced uveitis in rat eyes. They were more potent (at 3-10 mg/kg t.i.d.) than prednisolone (20 mg/kg t.i.d.) in effectively inhibiting posterior uveitis. They were also found to inhibit fibroblast-like corneal cells at 10-300 micrograms/ml concentrations and conjunctival cells at 1-30 micrograms/ml levels. The incorporation of leucine into corneal and conjunctival cells was either stimulated or unaffected by CK 126, indicating that the inhibition of cell growth has nothing to do with the protein synthesis. However, the incorporation of uridine into corneal and conjunctival cells was markedly inhibited by CK 126 at 3-30 micrograms/ml concentrations whereas the incorporation of thymidine into the cells was inhibited at a lesser extent than that of uridine. These results indicate that cell inhibition by CK 126 could be related mainly to the synthesis of mRNA and, to a lesser extent, to DNA synthesis.

Nasal administration of retinal antigens maintains immunosuppression of uveoretinitis in cyclosporin-A-treated Lewis rats: future treatment of endogenous posterior uveoretinitis?

PURPOSE: Current treatment of autoimmune endogenous posterior uveoretinitis (EPU) is limited by drug toxicity, unpredictable relapses on dose reduction and resistance to therapy. Administration of autoantigens via gastrointestinal or respiratory mucosa prior to antigen exposure induces immune hyporesponsiveness (mucosal tolerance) to further antigen sensitisation. In this study we assessed whether mucosal tolerance induction was possible after immunisation with retinal antigens in experimental autoimmune uveoretinitis (EAU) in animals that were short-term immunosuppressed with cyclosporin A (CsA) to determine whether mucosal administration of retinal antigens can maintain immunosuppression in sensitised and immunosuppressed individuals. METHODS: Female Lewis rats were immunised with retinal extract (RE) and then treated as follows. Group 1 received no specific therapy and served as control; group 2 were fed CsA from day 7 to day 20 post-immunisation; group 3 received inhalational tolerance therapy with RE in addition to CsA; tolerance therapy was continued after day 20 when CsA was stopped. Experiments varying the timing and dosage of both tolerising and immunising antigen were also performed, the details of which are described. Incidence, day of onset and clinical activity were recorded and histopathological assessment of intraocular inflammation, in particular the extent of autoimmune target-organ damage, was graded semiquantitatively. RESULTS: Compared with controls and group 2, group 3 showed both a marked delay in disease onset and a reduction in disease severity. This effect was both dose and dose-timing dependent. Tissue damage assessed in terms of preservation of rod outer segments was significantly less in group 3. CONCLUSIONS: The success of combination therapy, clinically, remains unknown at present but these results support continuing present clinical trials of mucosal tolerance therapy and in particular have future implications for either maintaining or inducing immunosuppression in autoimmune diseases in combination with present immunosuppressive therapies.

Tacrolimus-rapamycin combination therapy for experimental autoimmune uveoretinitis.

Tacrolimus and rapamycin both belong to a new family of immunosuppressants, immunophilin ligands, but the mechanisms by which they inhibit T cell activation are different. Therefore, we tested the immunosuppressive effects of combination therapy with low doses of tacrolimus and rapamycin on experimental autoimmune uveoretinitis (EAU) in rats. Male Lewis rats, immunized with S-antigen (S-Ag) were given intraperitoneal injection of the combined drugs for 14 days after the immunization with S-Ag. Effects were evaluated by clinical observations, histological examination and immune response. The combination therapy with tacrolimus (0.1 mg/kg per day) and rapamycin (0.03 mg/kg per day) achieved 100% suppression of clinical EAU and 66.7% suppression of histological EAU; tacrolimus combined with a higher dose of rapamycin (0.1 mg/kg per day) caused 100% suppression clinically and histologically. Therapy with either drug alone achieved only partial suppression: tacrolimus alone (0.1 or 0.2 mg/kg per day) or rapamycin alone (0.03-0.2 mg/kg per day). Doubling the dose of either drug produced only 16.7% suppression with rapamycin or 50% suppression with tacrolimus. The serum antibody levels to S-Ag and proliferative response of lymphocytes to S-Ag were also significantly suppressed by the combination therapy with low doses of tacrolimus and rapamycin.

Anti-tumor necrosis factor alpha therapy suppresses the induction of experimental autoimmune uveoretinitis in mice by inhibiting antigen priming.

PURPOSE: Experimental autoimmune uveoretinitis (EAU) serves as a model for several immune-mediated diseases that affect the eye in humans. Previous studies indicated that tumor necrosis factor alpha (TNF-alpha) has an important proinflammatory role in EAU and possibly in human uveitis. In this study, the authors investigated the effect of anti-TNF-alpha therapy on EAU in mice. METHODS: Experimental autoimmune uveoretinitis was induced in B10.A mice by immunization with interphotoreceptor retinoid-binding protein (IRBP). The mice were treated with 100 or 300 microliters rabbit antiserum or polyclonal antibodies to human TNF-alpha. The treatment spanned either the afferent or the efferent stage of EAU (days -1, 1, 3, 5, 7, or days 8, 10, 12, 14, 16, respectively). Control animals were injected with preimmune rabbit serum at the corresponding times or were not treated. Three weeks after immunization, EAU was assessed by clinical evaluation and by histopathology. Immunologic responses were assessed by delayed-type hypersensitivity (DTH), lymphocyte proliferation to IRBP, and relative abundance of IRBP-primed splenocytes. RESULTS: The treatment with rabbit anti-TNF-alpha serum significantly ameliorated disease when given during the afferent stage but had no effect when given during the efferent stage of EAU. The effect on DTH, lymphocyte proliferation, and abundance of antigen-reactive cells roughly paralleled the effect on disease. CONCLUSIONS: Neutralization of systemic TNF ameliorates EAU. The effectiveness of afferent treatment in comparison to the treatment during the efferent stage, together with the reduced proliferation and the reduced abundance of IRBP-responsive cells, suggest that interference with afferent-acting processes such as antigen priming is important to achieve protection from EAU by anti-TNF treatment.

Tetrandrine inhibits breakdown of blood-aqueous barrier induced by endotoxin and interleukin-1 alpha in rats.

Tetrandrine was shown to significantly inhibit uveitis induced by endotoxin and interleukin-1 alpha (IL-1 alpha) in rats. The dose-response curve of IL-1 alpha-induced uveitis was inhibited in a non-competitive manner. The maximum inflammation induced by IL-1 alpha was suppressed to 58.4%, 38.3% and 18.3% of the control peak by 5 mg/kg, 10 mg/kg and 20 mg/kg t.i.d. of tetrandrine, respectively. The maximum inflammation induced by endotoxin was suppressed to 56.5% and 38.0% by 5 mg/kg and 10 mg/kg t.i.d. of tetrandrine, respectively. The mechanism of tetrandrine's anti-inflammation could involve numerous pathways of inflammation processes and multiple inflammatory mediators. The results of this study indicate that tetrandrine appears to be a broad spectrum, non-steroidal, novel ocular anti-inflammatory agent.