A novel anti-mouse CD3ε antibody, Dow2, recognizes mouse CD3ε without activating T cells and suppresses T-cell activation. The purpose of this study was to determine whether Dow2 can inhibit T cells in uveitis.Experimental autoimmune uveitis (EAU) was induced in mice by immunization with retinal peptides, followed by administration of Dow2. Inflammation was evaluated by color fundus photography, optical coherence tomography, fluorescein angiography, and histology. Intraocular cells from EAU mice were used to examine the effect of Dow2 on retinal antigen-specific T cells. The effects of Dow2, conventional CD3ε antibodies, and isotype control immunoglobulin G (IgG) on splenic T cells were compared by assessing cell proliferation by the mixed lymphocyte reaction assay, inflammatory cytokine production by enzyme-linked immunosorbent assay (ELISA) and immunohistochemistry, and gene expression by quantitative reverse-transcription polymerase chain reaction (RT-PCR). T-cell subpopulations were characterized by flow cytometry to evaluate the expression of CD4, CD8, CD44, CD62L, and Foxp3.Dow2 significantly reduced T-cell activation and counteracted activation associated with anti-CD3ε antibodies. Unlike conventional CD3ε antibodies, Dow2 treatment did not upregulate T helper (Th)1-/Th17-associated gene expression and cytokine production in splenic T cells. Interferon (IFN)-γ production by retinal antigen-specific T cells was also significantly reduced. Ocular inflammation was significantly reduced in Dow2-treated EAU mice compared to control EAU mice, with fewer T cells infiltrating into the retinas of Dow2-treated EAU mice. In immunohistochemistry, Th1 and Th17 cells invaded the retina in control EAU mice but not Dow2-treated EAU mice. No effects on peripheral T-cell numbers were observed following systemic administration of Dow2.The novel anti-CD3 antibody Dow2 can inhibit T cell-mediated inflammation in uveitis models. Thus, inhibition of T-cell activation by anti-CD3 therapy with this new antibody may protect uveitis patients from severe ocular inflammation.

•Alloantibodies secreted by B cells target iPSC-derived RPE cells•Alloantibodies were detected in serum from monkeys receiving RPE allografts•Blood and lymph nodes in monkeys receiving the grafts contained activated B cells•B cell-mediated immune attack was attenuated by administration of local steroidAntibody-mediated rejection is characterized by donor-specific antibody produced by B cells. However, to our knowledge, B cell invasion and antibody in the inflamed retina after transplantation of retinal pigment epithelial (RPE) cells has not been reported. To determine if RPE transplantation could be performed using allografts, we established in vivo immune rejection models with induced pluripotent stem cell (iPSC)-RPE allografts and determined whether RPE-specific antibody could be detected in these models. We detected alloantibodies in the serum from recipient monkeys that had immune attacks in the retina in an immunofluorescent assay using the transplanted iPSC-RPE cells as the antigen. In addition to T cell and antigen-presenting cell immunity, peripheral blood cells and lymph nodes in animal models with allogeneic iPSC-RPE cells also had activated B cells, which were probably secreting alloantibodies. Using serum and transplanted cells, alloreactive antibody can be detected for the diagnosis of immune rejection after transplantation.Download high-res image (122KB)Download full-size image

To report a case of psoriasis triggered by anti-tumor necrosis factor-alpha (TNF-α) therapy in a uveitis patient with Behçet’s disease.A 34-year-old man with established Behçet’s disease was started on infliximab therapy for recurrent uveitis and showed an excellent response. After 2 years of infliximab treatment, he developed erythematous scaly plaques on both of his palms and heels. No clinical or serological evidence of infection was found, and there was no personal/family history of psoriasis. Histological examination of one lesion showed that it was consistent with psoriasis. Because of the development of hyperkeratotic skin lesions consistent with guttate psoriasis, the infliximab treatment was discontinued. Six months later, the psoriasis had resolved.Blockade of TNF-α is an effective treatment for psoriasis as well as Behçet’s disease. However, we have to consider new-onset psoriasis as an adverse side effect that can be triggered by anti-TNF-α therapy in Behçet’s disease.

To evaluate a broad-range real-time polymerase chain reaction (PCR) targeting the bacterial 16S rRNA gene for detection of bacterial DNA in infectious endophthalmitis.The bacterial 16S rRNA gene was measured by quantitative real-time PCR. For the assay, bacterial DNA was prepared from 12 Gram-positive and 4 Gram-negative strains. To determine the optimum method for DNA extraction, four extraction procedures were selected by using DNA extraction program cards with and without the use of lysozyme. To evaluate PCR sensitivity, PCR fragments were amplified from Staphylococcusaureus and Escherichiacoli DNA.DNA extraction using the Bacteria card® without enzymes resulted in detection of all the tested strains at concentrations ≥107 copies/mL. Extraction with the Bacteria card® with lysozyme resulted in detection of all the tested strains at concentrations ≥106 copies/mL, indicative of no significant difference between the two procedures. DNA extraction using the Virus card®, both with and without enzymes, resulted in reduced efficiency of detection of all strains compared with use of the Bacteria card®. The PCR could detect as few as 1–10 colony-forming units (CFU) in diluted vitreous samples per reaction, and all tested bacterial species known to cause endophthalmitis were detected.Bacterial 16S-specific PCR can comprehensively detect the main causative bacteria of clinically suspected endophthalmitis.