The Role of Interferon-γ in Autoimmune Polyendocrine Syndrome Type 1.

BACKGROUND: Autoimmune polyendocrine syndrome type 1 (APS-1) is a life-threatening, autosomal recessive syndrome caused by autoimmune regulator (AIRE) deficiency. In APS-1, self-reactive T cells escape thymic negative selection, infiltrate organs, and drive autoimmune injury. The effector mechanisms governing T-cell-mediated damage in APS-1 remain poorly understood. METHODS: We examined whether APS-1 could be classified as a disease mediated by interferon-γ. We first assessed patients with APS-1 who were participating in a prospective natural history study and evaluated mRNA and protein expression in blood and tissues. We then examined the pathogenic role of interferon-γ using Aire-/-Ifng-/- mice and Aire-/- mice treated with the Janus kinase (JAK) inhibitor ruxolitinib. On the basis of our findings, we used ruxolitinib to treat five patients with APS-1 and assessed clinical, immunologic, histologic, transcriptional, and autoantibody responses. RESULTS: Patients with APS-1 had enhanced interferon-γ responses in blood and in all examined autoimmunity-affected tissues. Aire-/- mice had selectively increased interferon-γ production by T cells and enhanced interferon-γ, phosphorylated signal transducer and activator of transcription 1 (pSTAT1), and CXCL9 signals in multiple organs. Ifng ablation or ruxolitinib-induced JAK-STAT blockade in Aire-/- mice normalized interferon-γ responses and averted T-cell infiltration and damage in organs. Ruxolitinib treatment of five patients with APS-1 led to decreased levels of T-cell-derived interferon-γ, normalized interferon-γ and CXCL9 levels, and remission of alopecia, oral candidiasis, nail dystrophy, gastritis, enteritis, arthritis, Sjögren's-like syndrome, urticaria, and thyroiditis. No serious adverse effects from ruxolitinib were identified in these patients. CONCLUSIONS: Our findings indicate that APS-1, which is caused by AIRE deficiency, is characterized by excessive, multiorgan interferon-γ-mediated responses. JAK inhibition with ruxolitinib in five patients showed promising results. (Funded by the National Institute of Allergy and Infectious Diseases and others.).

Preloading Trifolded Grafts for Descemet Membrane Endothelial Keratoplasty Affects Scroll Formation.

PURPOSE: The trifolded, endothelium-in approach to Descemet membrane endothelial keratoplasty (DMEK) facilitates tissue insertion into the anterior chamber. We hypothesized that preloading the trifolded donor grafts in a cartridge for 48 hours before insertion would induce biomechanical changes that decrease their scrolling tendency compared with those loaded immediately before insertion. METHODS: Ten Descemet membrane donor grafts, peeled and cut to 8.0 mm, were prepared by a single eye bank technician. Each graft was trifolded and pulled into a DMEK cartridge and stored for 48 hours. They were then pulled with microforceps into a petri dish filled with balanced salt solution. A video was recorded of the graft becoming a scroll over a 2-minute period. Each graft, serving as its own control, was then trifolded, pulled into the cartridge, and the process repeated. Images from 1, 5, 10, 60, and 120 seconds were extracted from video recording of the procedures. Scroll width was analyzed by graders masked to group assignment. A paired t test was used to determine differences in scroll width at each time point between the 48-hour and instant trifolding conditions. RESULTS: All grafts scrolled after removal from the cartridge into balanced salt solution. We measured a significant difference at all time points 1 through 120 seconds (4.02 preloaded vs. 2.91-mm instant trifold, P = 0.035). CONCLUSIONS: Preloading DMEK grafts in a trifolded configuration for 48 hours reduces the scrolling tendency of Descemet membrane for at least 2 minutes.

Qualitative and Quantitative Analysis of the Corneal Endothelium With Smartphone Specular Microscopy.

PURPOSE: We sought to demonstrate the feasibility of a lower-cost, portable method for qualitative and quantitative analysis of the corneal endothelium using a smartphone and slit-lamp biomicroscope. METHODS: In this study, at a single academic center, we recruited healthy participants to undergo imaging of the corneal endothelium using both a smartphone-based method and a specular microscope. Participants first had their eyes imaged with a CellChek NSP-9900 Specular Microscope (Konan Medical, Inc, Irvine, CA). For image capture on the smartphone, a beam of light approximately 0.2 mm in diameter was directed to the center of the cornea with a slit-lamp biomicroscope to achieve specular reflection. With 40× zoom on the slit-lamp and 4K video mode set on an iPhone 7 Plus held to an ocular, the corneal endothelium was recorded until the hexagonal pattern of cells was identified and the sharpest frame from the video was selected. RESULTS: The videos were analyzed from 14 sets of eyes (average length 2 minutes 40 seconds). The average intraclass correlation coefficient was 0.67 (95% confidence interval, 0.43-0.82). The mean difference between smartphone endothelial cell count and specular endothelial cell count was -209 cells/mm (SD = 483 cells/mm), which did not achieve significance (P = 0.14). A Bland-Altman analysis with simple linear regression showed no proportional bias when comparing the 2 modalities (coefficient = -0.20; t-value = -0.42; P = 0.68). CONCLUSIONS: Smartphone specular microscopy is capable of qualitative and quantitative analysis of the corneal endothelium. Further refinement to standardize the light source and automate analysis will increase feasibility.