Increased levels of monokine induced by interferon-gamma (Mig) in the vitreous of patients with diabetic retinopathy.

AIM: To determine the intravitreous concentration of monokine induced by interferon-gamma (Mig) in patients with diabetic retinopathy (DR) and the relation between Mig and vascular endothelial growth factor (VEGF). RESEARCH DESIGN AND METHODS: Vitreous samples were obtained at the time of vitrectomy from 41 eyes of 38 DR patients (30 with active DR and 11 with inactive DR) and from 15 eyes of 15 non-diabetic patients who had macular disease (control subjects). Human Mig and VEGF were quantified using a FACS Caliber flow cytometer. RESULTS: The vitreous concentration of Mig was increased significantly in patients with both active and inactive DR [148.0 (31.6-997.2; median range) and 82.3 (25.7-347.7) pg/ml, respectively] compared with control subjects [21.0 (5.2-74.3) pg/ml; P < 0.0001 and P < 0.001, respectively]. In DR patients, a significant (P < 0.01) correlation was observed between vitreous concentrations of Mig and VEGF. CONCLUSION: Our results suggest that Mig may play an important role in the pathogenesis of DR and works in consort with VEGF in the progression of pathological angiogenesis in DR.

A new method for assessing motion-in-depth perception in strabismic patients.

AIM: In strabismus clinics, stereoscopic depth perception is usually examined using static stimuli, but these stimuli do not necessarily allow assessment of the ability to perceive motion in depth. We assessed the ability to perceive motion-in-depth perception using a novel stereo motion test that we developed and compared with that to perceive static depth perception using a conventional stereo test in strabismic patients. METHODS: To investigate motion-in-depth perception in patients with strabismus, we developed a stereo motion test using four types of computer-generated dynamic visual stimuli. Three of them are random dot stereograms of two parallel planes moving in depth. The patient is asked to indicate the planes' direction of rotation in depth (in the first and second types) or the presence/absence of motion-in-depth signal (in the third type). The fourth type of stimulus was a random dot stereogram of a rotating cylinder. The upper and lower parts of the cylinder rotate in opposite directions, and the patient is asked to indicate the position of the border between the two parts. Threshold disparity was defined as the disparity (relative disparity between the nearest and farthest points of the planes or the cylinder) that gives a critical level of performance with the method of limit. The conventional Titmus stereo test using static visual stimuli was used to assess static depth perception. The measurements were performed in 52 strabismic patients, aged between 4 and 38 years old, who visited Tokyo Medical University Hospital between January 2003 and July 2004. RESULTS: The results showed a poor correlation in the threshold of individual patients between the stereo motion test and conventional Titmus stereo test. For example, the ability to perceive motion in depth (disparity threshold <500 sec of arc) was demonstrated in three of seven patients who were not able to perceive depth using static stimuli (0/9 for Titmus circle). These results suggest that the process of the dynamic element of binocular depth perception is preserved in some of the strabismic patients who lack static stereopsis. CONCLUSION: This study indicates the importance of testing motion-in-depth perception as well as static depth perception in assessing stereopsis in strabismic patients.

Supplementation of CD4+CD25+ regulatory T cells suppresses experimental autoimmune uveoretinitis.

AIMS: To investigate whether supplementation of natural CD4+CD25+ regulatory T cells ameliorates mouse experimental autoimmune uveoretinitis (EAU) induced by CD4+ T cell-dependent interphotoreceptor retinoid-binding protein (IRBP). METHODS: C57BL/6 mice were immunised with human interphotoreceptor retinoid-binding protein peptide 1-20 (IRBP(1-20)), and IRBP(1-20)-sensitised T cells were obtained. CD4+CD25+ T cells derived from naive mice were cocultured with IRBP(1-20)-sensitised T cells, and their proliferation responses and cytokine production were measured. In addition, CD4+CD25+ T cells were transferred intravenously into mice 7 or 15 days after immunisation with IRBP(1-20), and the severity of EAU and T cell proliferation responses were evaluated. RESULTS: CD4+CD25+ regulatory T cells effectively inhibited both the proliferation of, and interleukin (IL)2, IL5 and interferon (IFN)gamma production by, IRBP(1-20)-sensitised T cells. Adoptive transfer of CD4+CD25+ regulatory T cells to IRBP(1-20)-immunised mice conferred considerable protection from EAU development and inhibition of T cell proliferation responses to IRBP(1-20). CONCLUSION: These findings show that natural CD4+CD25+ regulatory T cells possess the ability to inhibit activation of IRBP-reactive T cells that have been already sensitised in vivo, and adoptive transfer of these cells ameliorates EAU even in the effector phase. Supplementation of natural CD4+CD25+ regulatory T cells may have therapeutic potential for effective treatment of uveitis.

Evidence for antigen-specific immune deviation in patients with acute retinal necrosis.

BACKGROUND: Because experimental acute retinal necrosis (ARN) induced by herpes simplex virus in mice develops only if mice fail to acquire virus-specific delayed hypersensitivity (DH), although they produce antiviral antibodies (ie, anterior chamber-associated immune deviation), we sought to determine whether a similar inverse correlation exists for patients with varicella-zoster virus (VZV)-induced ARN. DESIGN: Patients with acute, VZV-induced ARN and age-matched control subjects were skin tested with VZV and purified protein derivative antigens to evaluate DH. Varicella-zoster virus-induced ARN was diagnosed using polymerase chain reaction and intraocular antibody quotient. Serum samples were collected and analyzed for anti-VZV and anti-herpes simplex virus antibody titers. Acute retinal necrosis activity was assessed clinically, and DH skin tests were repeated 3 months after onset when ocular recovery had taken place. RESULTS: Whereas controls displayed intense DH when tested with VZV and purified protein derivative antigens, a subset of patients with ARN displayed absent VZV-specific DH (although their purified protein derivative responses were normal). Patients with the most severe ARN had the lowest DH responses to VZV antigens. Serum anti-VZV antibody titers were higher in patients with ARN than in controls, and antiviral titer correlated inversely with the intensity of anti-VZV DH responses. Varicella-zoster virus-specific DH responses were restored in patients who recovered from ARN. CONCLUSION: Varicella-zoster virus-ARN develops in a setting where DH reactivity to viral antigens is absent, implying that virus-specific DH might ameliorate the severity of ARN. CLINICAL RELEVANCE: Linking virus-specific DH to vulnerability to ARN in individuals infected with VZV might reveal an underappreciated pathogenic mechanism.

Participation of pigment epithelium of iris and ciliary body in ocular immune privilege. 2. Generation of TGF-beta-producing regulatory T cells.

PURPOSE: To determine whether T cells exposed to cultured iris and ciliary body pigment epithelial (I/CB PE) cells acquire the capacity to modify the activation, differentiation, and effector functions of bystander T cells, and if so, to identify the mechanism. METHODS: T cells from naive BALB/c mice were cultured with I/CB PE cells, x-irradiated, and used as regulators (a) of T-cell activation in vitro and (b) of delayed hypersensitivity expression in vivo. Neutralizing anti-TGF-beta and -IL-10 antibodies were used to abolish regulatory function. T-cell activation was assessed for proliferation by [(3)H]thymidine incorporation and for IL-2, IFN-gamma, IL-4, and IL-10 production by semi-quantitative RT-PCR for mRNA and by supernatant analysis by ELISA. I/CB PE-exposed T cells were evaluated for mRNA content of IFN-gamma, IL-4, TNF-alpha, TGF-beta1, TGF-beta2, and IL-10, and their supernatants were analyzed for content of TGF-beta. RESULTS: T cells exposed to I/CB PE cells inhibited anti-CD3-driven activation of bystander naive T cells in vitro and suppressed the expression of delayed hypersensitivity in vivo. Bystander T cells cocultured with I/CB PE-exposed T cells failed to proliferate and secreted high levels of IL-4 and IL-10 but low amounts of IL-2 and IFN-gamma. Regulation of bystander T-cell activation was mediated via enhanced secretion of TGF-beta by I/CB PE-exposed T cells. CONCLUSIONS: T cells exposed to cultured I/CB PE cells were induced to secrete active and latent TGF-beta, which conferred on the T cells the capacity to inhibit the differentiation as well as the effector function of Th1-type cells.

In vitro generation of regulatory CD8+ T cells similar to those found in mice with anterior chamber-associated immune deviation.

PURPOSE: When injected intravenously into naive mice, peritoneal exudate cells (PECs) incubated with ovalbumin (OVA) in the presence of transforming growth factor (TGF)-beta2 induce immune deviation similar to that evoked by injection of OVA into the anterior chamber of the eye. Intraocular antigen injection elicits two distinct populations of regulatory T cells that impair delayed hypersensitivity (DH) by two different mechanisms: a CD4+ T cell that suppresses the induction of DH (afferent) and a CD8+ T cell that inhibits DH expression. In an effort to understand the origin and mechanism of action of these regulatory cells, CD8+ T cells from OVA-specific T cell receptor (Tcr) transgenic mice (OT-1) were used. METHODS: CD8+ T cells were harvested from Tcr transgenic OT-1 mice whose Tcr recognize an OVA peptide in the context of the class I major histocompatibility complex molecule Kb. These cells were stimulated in vitro with OVA-pulsed PECs exposed (or not) to TGF-beta2, then analyzed for their capacity to proliferate, to secrete various cytokines, to lyse OVA-expressing target cells, and to regulate bystander T cells in vitro and in vivo. RESULTS: When OVA-pulsed PECs were used in vitro as stimulators, responding OT-1 T cells proliferated and preferentially secreted interferon (IFN)-gamma, interleukin (IL)-2, and tumor necrosis factor (TNF)-alpha, rather than IL-4 and IL-10. When the stimulator PECs were pretreated with TGF-beta2 and then pulsed with OVA, responding OT-1 T cells proliferated even more swiftly, but they secreted significantly less IFN-gamma, IL-2, and TNF-alpha, and no IL-4 or IL-10. OT-1 T cells, which constitutively display cytotoxicity toward OVA-expressing target cells, lost this activity when stimulated with OVA-pulsed, TGF-beta2-pretreated PECs. Moreover, OT-1 T cells stimulated in this manner displayed the capacity to inhibit proliferation of OVA-primed T cells exposed to OVA in vitro and to suppress in vivo the expression of OVA-triggered DH. CONCLUSIONS: OVA-pulsed PECs, pretreated with TGF-beta2, coerce naive OVA-specific CD8+ T cells to become efferent regulators of DH similar to the regulatory T cells evoked by intraocular injection of OVA.

Analysis of in vivo regulatory properties of T cells activated in vitro by TGFbeta2-treated antigen presenting cells.

PURPOSE: To determine whether naive T cells activated in vitro by antigen-pulsed, transforming growth factor(beta) (TGFbeta)-treated antigen presenting cells (APCs) acquire the capacity to suppress the induction and expression of delayed hypersensitivity in vivo. METHODS: Naive ovalbumin (OVA)specific T cells from DO 11.10 Tcr transgenic mice were stimulated in vitro with OVA-pulsed TGF(beta2)-treated APCs. The cultured cells were harvested and assayed for in vitro production of mature TGFbeta. Similar cells were coinjected with primed OVA-specific BALB/c T cells plus OVA-pulsed APCs into ear pinnae of normal BALB/c mice (assay for delayed hypersensitivity expression) or coinjected with OVA-pulsed APCs into footpads of naive DO11.10 mice whose draining lymph node cells were harvested 4 days later and assayed in vitro for capacity to secrete interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) when stimulated with OVA (assay for induction of delayed hypersensitivity). RESULTS: DO11.10 T cells activated in vitro by OVA-pulsed TGFbeta2-treated APCs secreted large amounts of mature TGFbeta and suppressed the expression of delayed hypersensitivity in a local adoptive transfer assay. Suppression was reversed in the presence of neutralizing anti-TGFbeta antibodies. In addition, in vitro generated regulatory T cells influenced naive T cells in DO11.10 mice that were responding to an initial immunization with OVA to secrete IL-4, rather than IFN-gamma. This influence was independent of TGFbeta. CONCLUSIONS: OVA-pulsed APCs, pretreated in vitro with TGF(beta)2, activate DO11.10 T cells in a manner that endows the responding cells with the capacity to suppress the induction and then the expression of delayed hypersensitivity in vivo. In certain ways, these properties of in vitro-activated DO11.10 T cells resemble the properties of afferent and efferent regulatory T cells typically found in the spleens of animals with anterior chamber-associated immune deviation.

Evidence for multiple CD95-CD95 ligand interactions in anteriorchamber-associated immune deviation induced by soluble protein antigen.

We have investigated whether CD95-CD95 ligand interactions are important in anterior chamber-associated immune deviation (ACAID) induced by soluble protein antigen, and if so, to identify the participating cells on which these molecules are expressed. Peritoneal exudate cells as antigen-presenting cells (APC) obtained from B6.lpr/lpr, B6.gld/gld and C57BL/6 mice were cultured with ovalbumin (OVA) and transforming growth factor-beta2 (TGF-beta2) overnight, then injected intravenously into C57BL/6 or B6.lpr/lpr recipients. Some B6.lpr/lpr mice were reconstituted with naive T cells from wild-type C57BL/6 donors. In other experiments, B6. lpr/lpr and B6.gld/gld mice received an anterior chamber injection of OVA followed 7 days later by subcutaneous immunization with OVA plus adjuvant. Delayed hypersensitivity (DH) was assessed with an ear swelling assay. T cells activated in vitro with OVA-pulsed, TGF-beta-treated APC were tested in vivo for their capacity to suppress DH expression in a local adoptive transfer assay. The results indicate that when ACAID was induced by in-vitro generated ACAID-inducing cells, the APC expressed CD95L, and recipient T cells expressed CD95. The capacity of in vitro generated regulatory T cells to suppress DH expression to OVA in vivo was not governed by CD95-CD95L interactions. When OVA was injected into the anterior chamber of naive mice, CD95 expression was required for ACAID induction, although ACAID was readily induced in CD95L-deficient mice. We conclude that CD95-CD95L interactions are required in ACAID for the initial stage of APC presentation of eye-derived antigens to T cells, and that CD95-CD95L interactions participate at one or more additional step in the process by which ACAID is induced by soluble protein antigens.

The role of costimulatory molecules B7-1 and B7-2 in mice with experimental autoimmune uveoretinitis.

BACKGROUND: Onset of experimental autoimmune uveoretinitis (EAU) is believed to involve a CD4-positive type 1 T helper cell (Th1) immune response, with inhibition involving a Th2 immune response. Development of Th1 and Th2 responses involves the participation of the costimulatory molecules B7-1 and B7-2, respectively. The purpose of this study was to investigate the role of B7-1 and B7-2 in the EAU model in mice. METHODS: B10.A mice were immunized with interphotoreceptor retinoid-binding protein (IRBP) and given daily intraperitoneal injections of either phosphate-buffered saline (control), mouse monoclonal antibody (mAb) to B7-1, mAb to B7-2, or mAb to both B7-1 and B7-2. Eyes were evaluated by histopathological criteria and cytokines were assayed in culture medium of IRBP-stimulated lymphocytes. Cellular immune responses were measured by cell proliferation assay under IRBP stimulation. RESULTS: Rates of EAU onset were 5/10 (50%) for control mice, 1/9 (11%) for mice treated with anti-B7-1 mAb, 5/6 (83%) for mice treated with anti-B7-2 mAb, and 2/6 (33%) for mice treated with both anti-B7-1 and anti-B7-2 mAb. Mean histopathological severity scores were 2. 4+/-0.8, 1.0+/-0, 2.6+/-1.0, and 1.0+/-0, respectively. Production of IL-5 was significantly increased in mice treated with anti-B7-1 mAb, while IFN-gamma was increased in mice treated with anti-B7-2 mAb. Spleen cell proliferation was significantly reduced in mice treated with anti-B7-1 mAb. CONCLUSIONS: These results suggest that the costimulatory molecules B7-1 and B7-2, via their influence on generating Th1 and Th2 immune responses, play an important role in the clinical outcome of EAU in mice immunized with IRBP.

Conserved T cell receptor complementarity-determining region 3 of ocular T cells in mice with experimental autoimmune uveoretinitis.

PURPOSE: To characterize T cells infiltrating into the ocular tissues of mice with experimental autoimmune uveoretinitis (EAU) immunized with interphotoreceptor retinoid-binding protein (IRBP). METHODS: The T cell receptor (TCR) repertoire on T cells obtained from ocular lesions of EAU mice was analyzed by reverse transcriptase-polymerase chain reaction. The clonotype of the T cells was examined by the single-strand conformation polymorphism (SSCP) method, followed by sequence analysis of the TCR beta-chain complementarity-determining region 3 (CDR3). RESULTS: The repertoire of the TCR BV gene in T cells from inflamed lesions was heterogeneous. SSCP analysis showed accumulation of multiple T cells specifically in ocular tissues of EAU mice, suggesting that these cells were expanded by an antigen-driven stimulation. Junctional sequence analysis demonstrated the presence of highly conserved amino acid sequence motifs (AGTGG, AGD) in CDR3 of BV2-positive T cells. CONCLUSIONS: Our findings suggest that T cells infiltrating into ocular lesions of EAU mice recognize restricted T cell epitopes of IRBP, resulting in autoimmune uveoretinitis.

[Suppression of experimental autoimmune uveoretinitis with a combination of cyclosporin and allopurinol].

PURPOSE: We assessed the suppressive effect of a combination of cyclosporin (an immunosuppressive agent) and allopurinol (a xanthine oxidase inhibitor and radical scavenger) on experimental autoimmune uveoretinitis (EAU) in Lewis rats, induced by interphotoreceptor retinoid-binding protein (IRBP). METHODS: After the immunization of Lewis rats with 30 micrograms of IRBP. We administrated cyclosporin and/or allopurinol to the IRBP-immunized Lewis rats. We observed the incidence and the severity of EAU. Histological, immunological, and biochemical examinations were performal 13 days after the immunization. The suppressive effect of these drugs in vitro on the production of free radicals derived from polymorphonuclear leukocytes. RESULTS: The incidence of EAU was suppressed by 50% at 13 days after immunization, and in terms of clinical and histological findings, inflammatory reaction was more inhibited by the combination of these drugs than by either cyclosporin or allopurinol alone. Lymphocyte proliferation assay against IRBP was significantly inhibited by the combination of drugs. No adverse systemic effects were identified. Cyclosporin and allopurinol inhibited radical production both separately and in combination. CONCLUSION: This suggests that suppression of EAU is based not only on inhibited cell-mediated immunity but also on inhibited production of free radicals derived from polymorphonuclear leukocytes.

[Association of heat-shock protein and uveitis].

Heat-shock protein 60 derived from Yersinia enterocolitica (Yersinia HSP 60) and bovine retinal HSP (Retina HSP 60) were previously identified by immunological cross-reaction and a high degree of common antigenicity, and a specific antibody against both proteins was detected in the sera of uveitis patients. We report here an attempt to isolate and purify Retina HSP 60 and Yersinia HSP 60. Both Retina HSP 60 and Yersinia HSP 60 showed an enriched content of glycine of approximately 60 to 80%. Lewis rats were inculated with 50 or 100 micrograms of purified Retina or Yersinia HSP 60 emulsified in complete Freund's adjuvant. In 50 to 60% of those inoculated with Retina HSP 60, uveoretinitis was observed about 13 days after inoculation, with massive infiltration of lymphocytes and polymorphonuclear neutrophils in the iris, ciliary body, and retinal tissue. Rats inoculated with Yersinia HSP 60 did not develop ocular inflammation. Lymphocyte proliferation assay was performed to investigate cellular immunoresponses in the rats that developed ocular inflammation after immunization with Retina HSP 60. The results showed significantly higher response to the Retina and Yersinia HSP 60 than to either S-antigen or interphotoreceptor retinoid-binding protein (IRBP), which are known to induce ocular inflammation. Cross-reaction between Retina and Yersinia HSP 60 is suggested. This study suggests that the HSP 60 molecules may be involved in the pathogenesis of intraocular inflammation.

Suppression of spontaneous uveoretinitis development by non-immunopathogenic peptide immunization.

BALB/c nude mice which are grafted with thymus tissue from fetal F344 rats beneath the renal capsule (hereafter referred to as TG nude mice) spontaneously develop uveoretinitis as well as other organ-localized autoimmune diseases. Active immunization with an interphotoreceptor retinoid-binding protein (IRBP)-derived peptide, amino acids 518-529 (P518-529), induced rapid development and high incidence of uveoretinitis, whereas immunization with another amino acid fragment, 1182-1194 (P1182-1194), inhibited the disease process. P1182-1194- or P518-529-specific T cell lines were established from TG nude mice. Although both were of CD4+ type, P518-529-specific T cells expressed Vbeta8 TCR while Vbeta6 expression was evident in the P1182-1194-specific cells. P518-529-specific T cells produced IL-2 and IFN-gamma, but not IL-4 or IL-10, whereas P1182-1194-specific T cells produced IL-4 and IL-10, but not IL-2 or IFN-gamma Adoptive transfer of these peptide-specific T cells into naive BALB/c nude mice resulted in development of uveoretinitis only in the P518-529 case. Furthermore, mice receiving both T cell types simultaneously did not exhibit uveoretinitis. The results indicate that the amino acid fragment of IRBP, P518-529, is uveitogenic and immunogenic in TG nude mice and induces Th1-type T cells related to uveoretinitis, whereas the amino acid fragment 1182-1194 is immunogenic but not uveitogenic, inducing Th2-type T cells which are involved in inhibition of this pathological response in TG nude mice.

[Inhibition of experimental autoimmune uveoretinitis by transforming growth factor-beta 1 in B10. A mice].

Experimental autoimmune uveoretinitis (EAU) in mice, an organ specific autoimmune disease, has been investigated as an animal model for human endogenous uveitis. In this study, we report on the immunosuppressive effect of transforming growth factor-beta 1 (TGF-beta 1) on the development of EAU in mice. Inhibition by TGF-beta 1 of proliferation of interphotoreceptor retinoid-binding protein (IRBP)-specific T cell lines in B10.A mice against IRBP antigen was dose-dependent. However, when spleen cells used as the antigen presenting cell were first cultured with TGF-beta 1, this anti-proliferation effect was abolished. When IRBP-immunized mice were injected intraperitoneally with TGF-beta 1, dose-dependent suppression of EAU was obtained. The proliferation response of lymph node cells from TGF-beta 1 injected mice with IRBP-induced EAU was suppressed compared with phosphate buffered saline (PBS)-injected mice. These findings suggest that TGF-beta 1 may be a cytokine that plays a role in suppressing IRBP induced EAU in mice.

Prevention of experimental autoimmune uveoretinitis by monoclonal antibody to interleukin-12.

Experimental autoimmune uveoretinitis (EAU) induced by immunization with interphotoreceptor retinoid-binding protein (IRBP), a retinal self antigen, has been regarded to be a typical T helper type 1 (Th1)-mediated inflammatory disease. In this study, we examined the effect of a neutralizing monoclonal antibody (mAb) to interleukin-12 (IL-12), which has been known to play a critical role in the Th1 differentiation, on the development of EAU. While 9 of 13 control mice developed EAU by the immunization with IRBP, none of 12 mice developed EAU when given anti-IL-12 mAb 1 day before immunization. These mice did not develop EAU even after a rechallenge with IRBP on day 30, indicating that a protective mechanism had been established by the anti-IL-12 treatment. The proliferative response of splenocytes to IRBP in vitro was not significantly impaired, but the production of IL-2 and IFN-gamma was greatly reduced by the anti-IL-12 treatment. Moreover, production of IL-5 and expression of IL-4 mRNA were increased by the anti-IL-12 treatment. Consistently, IgG2a anti-IRBP serum antibodies were decreased and IgG1 were increased. Administration of a neutralizing anti-IL-4 mAb at the time of IRBP rechallenge reversed the protection established by the anti-IL-12 treatment at the primary immunization. These results indicate that the anti-IL-12 treatment at the IRBP priming not only prevented the development of pathogenic Th1 cells, but also induced suppressive Th2 cells that protect the animals from further challenge with the same antigen.

Peptide-mediated suppression of experimental autoimmune uveoretinitis in mice: development of a peptide vaccine.

Experimental autoimmune uveoretinitis (EAU) is an animal model of antigen-specific, Th cell-mediated, organ-specific autoimmune disease. EAU is induced by immunization of B10.A mice with interphotoreceptor retinoid-binding protein (IRBP). Pre-treatment with synthetic peptide 518-529 derived from IRBP prevented IRBP-mediated EAU. This was accompanied by augmentation of the IRBP-specific IgG1 antibody (Th2) response and down-regulation of the IRBP-specific IgG2a (Th1) response. Consistent with this is the observation that two of two T cell lines established from p518-529-primed mice produced Th2-type cytokines (IL-4 and IL-10), whereas three of three T cell lines obtained from IRBP-primed mice produced Th1-type cytokines (IL-2 and IFN-gamma). Together this suggests the possibility that p518-529 priming causes a shift from a Th1-to a Th2-dominated immune response, thereby playing a pivotal role in the prevention of IRBP-mediated EAU. Furthermore, co-transfer of cells from a CD4+ p518-529-specific T cell line prevented the development of EAU after adoptive transfer of spleen cells from mice with EAU into normal mice. These findings contribute to our understanding of the mechanism of EAU, particularly with respect to the down-regulation of Th1-initiated inflammation, and may prove valuable for designing a peptide vaccine for EAU in the future.

Tissue-specific suppressor T cells involved in self-tolerance are activated extrathymically by self-antigens.

Autoimmune prostatitis developed spontaneously in (C57BL/6N x A/J)F1 (B6A) mice, when thymectomy (Tx) was conducted on day 3 after birth (Tx-3). The lesion could be prevented by a single injection of CD4+ spleen cells (4 x 10(6)) from normal males, but not from normal females or newborn orchidectomized (Orx-0) mice. However, when spleen cells were obtained from Orx-0 mice that had received a dihydrotestosterone (DHT) pellet when adult to develop a mature prostate, prostatitis could be prevented, suggesting that immune tolerance to prostate antigen(s) is maintained by a CD4+ tissue-specific suppressor T cell (Ts) population(s), which is activated by a specific autoantigen(s) in the mature prostate. The result that even CD4+ cells from Orx-0 mice that were thymectomized as adults and treated thereafter with DHT were effective for prevention of prostatitis suggests that activation of this Ts population takes place in the peripheral lymphoid organs, and that it maintains peripheral tolerance to autoantigen in the prostate of these mice and probably also in normal mice.