ovalbumin and Uveitis--Anterior

To examine antigen (Ag)-specific CTL response during anterior chamber associated immune deviation (ACAID).. OVA or OVA257-264 peptide was injected into the anterior chamber (AC) of C57BL/6 mice. There were 16 mice in each ACAID group induced with OVA or OVA257-264 peptide. The mice were primed by SC injection with OVA or OVA 257-264 peptide in complete Freund's adjuvant (CFA) on day 7. Ag-specific CD8+ T cells in spleens were analyzed on day 14 using Pentamer H-2K(b)-SIINFEKL(OVA257-264 peptide). IFN-gamma ELISPOT and intracellular granzyme B staining were used to characterize the CTL response. Twelve mice in each group immunized with OVA or OVA257-264 peptide in CFA served as positive controls. Twelve normal mice served as negative controls and 12 receiving injection of CFA as CFA controls for studying the influence of CFA on the Ag-specific CTL response.. The results showed that anterior chamber inoculation of OVA or OVA257-264 peptide could induce ACAID as evidenced by an impaired DTH response. The frequency of Ag-specific CD8+ T cells in ACAID mice was not different from that in mice challenged with Ags in CFA only (positive controls). IFN-gamma production by these cells in ACAID mice was not different compared to positive controls. However, Ag-specific CD8+ T cells in ACAID mice failed to secrete granzyme B. Mice challenged only with OVA peptide and CFA also showed a granzyme B negative CD8+ T cell response. Ag-specific CTL response induced by CFA alone was similar with the negative control.. These results show that the frequency of Ag-specific CD8+ T cells is not altered during ACAID. The Ag-specific CTL response during ACAID is characterized by the absence of granzyme B expression.

Topics: Animals; Anterior Chamber; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Freund's Adjuvant; Granzymes; Hypersensitivity, Delayed; Immunization; Interferon-gamma; Mice; Mice, Inbred C57BL; Ovalbumin; Peptide Fragments; Specific Pathogen-Free Organisms; Spleen; T-Lymphocyte Subsets; T-Lymphocytes, Cytotoxic; Uveitis, Anterior

Uveitis is an immune-mediated ocular disease and a leading cause of blindness. We characterized a novel model of uveitis with intravital microscopy. Transfer of ovalbumin-specific T cells from DO11.10 spleen to BALB/c recipients and subsequent challenge with ovalbumin in the anterior chamber of the eye resulted in anterior uveitis. Antigen-specificity was verified by injection of irrelevant antigen and transfer of T cells with a different specificity. Subsets of CD4 T cells, including naive (DO11.10 RAG(-/-)) and in vitro-activated Th2 effector CD4 T cells, infiltrated anterior segment tissues early in the inflammation. Memory-like CD44(high) CD4 T cells from unprimed transgenic mice and in vitro-activated Th1 effector CD4 T cells accumulated to larger numbers than naive or Th2 effector cells at 48 and 72 h. Of these, the alpha(2)-integrin+CD4 unprimed T cells entered the eye more efficiently, and antibody to alpha(2)-integrin markedly inhibited the inflammatory response. Intravital microscopy revealed the early arrival and antigen-specific accumulation of CD4 T cells in inflamed tissue and should be helpful in understanding T cell migration to other organs.

Topics: Animals; Antigens; CD4-Positive T-Lymphocytes; Cell Count; Female; Gene Expression Regulation; Hyaluronan Receptors; Immunity, Innate; Immunologic Memory; Integrin alpha2; Mice; Mice, Inbred BALB C; Mice, Knockout; Microscopy; Ovalbumin; Uveitis, Anterior

To study the expression and functional characteristics of programmed death-1 (PD-1) and its ligands in the spleens of mice undergoing anterior chamber-associated immune deviation (ACAID).. ACAID was induced in BALB/c mice by intracameral injection of ovalbumin (OVA). The expression of PD-1 and its ligands in the spleens of ACAID mice was determined by quantitative real-time PCR, Western blotting, and flow cytometry. In vitro proliferation assays, enzyme-linked immunosorbent assays, and adoptive transfer assays were used to investigate the functional characteristics of splenic CD4+PD-1+ T cells of ACAID mice.. Both mRNA and protein of PD-1, PD-L1, and PD-L2 were markedly upregulated in the spleens of ACAID mice compared with controls. CD4+PD-1+ T cells from ACAID mice produced large amounts of IL-10 and exhibited in vitro antigen-specific suppressive activity. CD4+PD-1+ T cells from ACAID mice were able to significantly inhibit the antigen-specific, delayed-type hypersensitivity response when adoptively transferred to naive mice.. CD4+PD-1+ T cells from ACAID mice, as regulatory cells, are involved in the induction of antigen-specific suppression in association with enhanced expression of IL-10. CD4+PD-1+ T cells in the murine spleen may represent a substantial population of regulatory T cells possibly responsible for the induction of ACAID after intracameral injection of antigen.

Topics: Adoptive Transfer; Animals; Anterior Chamber; Antigens, Surface; Apoptosis Regulatory Proteins; B7-1 Antigen; B7-H1 Antigen; Blotting, Western; CD4 Antigens; Disease Models, Animal; Enzyme-Linked Immunosorbent Assay; Female; Flow Cytometry; Hypersensitivity, Delayed; Interleukin-10; Lymphocyte Activation; Membrane Glycoproteins; Mice; Mice, Inbred BALB C; Ovalbumin; Peptides; Phenotype; Programmed Cell Death 1 Receptor; Reverse Transcriptase Polymerase Chain Reaction; RNA, Messenger; Spleen; T-Lymphocytes, Regulatory; Up-Regulation; Uveitis, Anterior

To determine whether an increased or reduced inflammatory response following cataract surgery influences the development of after-cataract.. Department of Preclinical Ophthalmology, Pharmacia, Uppsala, Sweden.. Rabbits that had had cataract surgery were given endotoxin, ovalbumin, dexamethasone, or diclofenac. Aqueous humor, leukocytes, and prostaglandin E(2) (PGE(2)) were analyzed, and the wet weight of the after-cataract was determined.. The wet weight of the after-cataract was unaffected by endotoxin and 67% lower in the eyes treated with ovalbumin than in the control eyes on day 56. Aqueous humor concentrations of leukocytes and PGE(2) were 94% and 87% lower in the group treated with dexamethasone than in the control group on day 7, and the concentration of PGE(2) was 98% lower in the diclofenac group; however, the wet weight of the after-cataract was unaffected by both treatments.. This study suggests that an increased inflammatory response does not increase the development of after-cataract and a reduction in the inflammatory response does not reduce the development of after-cataract.

Topics: Animals; Anti-Inflammatory Agents, Non-Steroidal; Aqueous Humor; Cataract Extraction; Dexamethasone; Diclofenac; Dinoprostone; Endotoxins; Glucocorticoids; Leukocytes; Ophthalmic Solutions; Ovalbumin; Pilot Projects; Rabbits; Uveitis, Anterior