Ocular infiltrating CD4+ T cells from patients with Vogt-Koyanagi-Harada disease recognize human melanocyte antigens.

PURPOSE: To determine whether patients with Vogt-Koyanagi-Harada (VKH) disease have immune responses specific to the melanocyte antigens tyrosinase and gp100. METHODS: T-cell clones (TCCs) were established from cells infiltrating the aqueous humor and from peripheral blood mononuclear cells (PBMCs) of patients with VKH. The target cells were LDR4-transfected cells (HLA-DRB1*0405). The TCCs were cocultured with LDR4 in the presence of tyrosinase (tyrosinase450-462: SYLQDSDPDSFQD), gp100 (gp100(44-59): WNRQLYPEWTEAQRLD), or a control peptide. The immune response was evaluated by cytokine production. The responding melanocyte peptide-specific VKH-TCCs were characterized by an immunofluorescence method with flow cytometry. A search was made for molecular mimicry among tyrosinase450-462, gp100(44-59), and exogenous antigens, such as viruses, by database screening. RESULTS: Cells infiltrating the eye and PBMCs in HLA-DR4+ (HLA-DRB1*0405, 0410) patients with VKH contained a population of CD4+ T lymphocytes that recognized tyrosinase and gp100 peptides and produced RANTES and IFN-gamma in response to the two peptides. The T cells were active memory Th1-type lymphocytes, and they recognized the tyrosinase peptide and produced IFN-gamma in response to HLA-DRB1*0405+ melanoma cells. Cytomegalovirus envelope glycoprotein H (CMV-egH290-302) had high amino acid homology with the tyrosinase peptide. In addition, some of the VKH-TCCs recognized CMV-egH290-302 peptide, as well as the tyrosinase peptides. CONCLUSIONS: In VKH there are tyrosinase and gp100 peptide-specific T cells that can mediate an inflammatory response. Such melanocyte antigen-specific T cells could be associated with the cause and pathology of VKH disease.

Cytokine profile in aqueous humor and sera of patients with infectious or noninfectious uveitis.

PURPOSE: To determine the cytokine expression profile at the protein level in aqueous humor (AqH) and sera of patients with uveitis. METHODS: Patients with various clinical entities of strictly diagnosed infectious or noninfectious uveitis were tested. AqH and sera were collected from patients with uveitis. AqH was also collected during surgery from patients with cataract, as control specimens. Interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, and interleukin (IL)-2, -4, -5, and -10 were measured from nondiluted samples simultaneously, with microparticle-based flow cytometric analysis. RESULTS: In AqH IFN-gamma was the most abundant cytokine in both infectious (mean, 3240.5 pg/mL) and noninfectious (mean, 115.6 pg/mL) uveitis, and IL-10 was the second (mean, 402.1 pg/mL, infectious uveitis; 7.5 pg/mL, noninfectious uveitis). The expression level of other cytokines in AqH was generally higher in infectious uveitis than in noninfectious uveitis, but the levels were lower than that of IL-10. There was no remarkable difference, however, in the cytokine expression pattern in AqH of the different clinical entities of uveitis. Sera from patients with noninfectious uveitis contained IFN-gamma (mean, 45.0 pg/mL), but the other serum cytokines in both types of uveitis were low or under the detectable level. CONCLUSIONS: IFN-gamma is the most abundant cytokine in infectious and noninfectious uveitis, with a remarkable difference between the two groups. The data suggest that cytokines in AqH of infectious uveitis are locally produced, whereas in noninfectious uveitis, IFN-gamma is produced both in the eye and the peripheral blood.

[A case of intraocular malignant lymphoma diagnosed by immunoglobulin gene rearrangement and translocation, and IL-10/IL-6 ratio in the vitreous fluid].

BACKGROUND: The diagnosis of primary intraocular lymphoma is difficult in many cases even with conventional cytological tests using vitreous samples. Recently new diagnostic tests, such as microdissection and polymerase chain reaction (PCR) and measurement of cytokines using intraocular samples, have been applied to the diagnosis of the disease. We report here a case where we used the new diagnostic tests and the results aided us to make a diagnosis of intraocular lymphoma. CASE: A 68-year-old woman with an initial diagnosis of bilateral idiopathic uveitis with steroid-resistant vitreous opacities underwent a vitreous biopsy. The cytological examinations of the vitreous samples revealed class III. The microdissection and PCR using the vitreous samples detected IgH rearrangement gene in the third framework (FR3A), the complementary determining region 3 (CDR3) of the VH region and Bcl-2-associated translocation. The interleukin (IL)-10 to IL-6 ratio in the vitreous fluid was greater than 100. Because the results of the examinations strongly suggested intraocular lymphoma, the patient was treated with radiation and chemotherapy. One month after the therapy, however, the patient developed multiple metastatic lesions in the brain. The clinical course of the patient together with the new diagnostic results of examinations led to a diagnosis of intraocular lymphoma. CONCLUSION: A combination of tests, such as conventional cytology, microdissection, and PCR, and cytokine assay using intraocular biopsy samples, is useful to make a diagnosis of intraocular lymphoma.

The presence of macrophage migration inhibitory factor in human trabecular meshwork and its upregulatory effects on the T helper 1 cytokine.

PURPOSE: To investigate the expression and secretion of macrophage migration inhibitory factor (MIF) in human trabecular meshwork (HTM) and evaluate its role in ocular inflammation. METHODS: Tissue samples of HTM cells were isolated from donor human eyes or corneoscleral buttons, and the HTM cells were cultured. The expression of MIF on HTM cells was evaluated by RT-PCR, Western blot analysis, and ELISA. T-cell clones (TCCs) were established from ocular infiltrating cells of patients with uveitis. ELISA was used to evaluate the pathologic role of MIF, in relation to regulatory effects on cytokine production by T cells. RESULTS: MIF was detected in the HTM by RT-PCR and Western blot analysis. MIF was also shown by ELISA to be secreted by the HTM cells in culture. The HTM supernatant enhanced IFN-gamma production by TCCs, but not IL-10; and these effects were neutralized by anti-MIF antibodies. Similarly, recombinant MIF enhanced the IFN-gamma production by the TCCs. CONCLUSIONS: MIF is expressed and secreted in the HTM, and MIF has the capacity to enhance T helper 1 cytokines and may play a role as an inflammatory cytokine in the eye.