DOI: 10.5061/dryad.k6djh9w8z
title: Ocular graft vs host disease mouse model

description: Ocular graft versus host disease (OGvHD) develops in nearly 50% of
 patients after allogeneic hematopoietic stem cell transplantation (HSCT)
 and frequently manifests as dry eye disease (DED).  The
 aim of this study was to evaluate the efficacy of a single subconjunctival
 (SCJ) dose of adeno-associated virus (AAV) gene therapy encoding human
 leukocyte antigen G (HLA-G) isoforms 1 and 5 to induce ocular immune
 tolerance and inhibit development of clinical signs of OGvHD.
description:  A Major histocompatibility mismatch chronic OGvHD murine model
 was evaluated. Seven days after HSCT, mice were dosed subconjunctivally
 with scAAV8-HLA-G1/5 (1e9vg/eye), topical cyclosporine (CSA) (twice
 daily), or left untreated.  Body weights and tear production (red thread
 test) were recorded, and eyelid, corneal opacity, and corneal fluorescein
 retention were scored through day 44 after HSCT. Body
 weights were recorded prior to ocular injections and every other day
 thereafter. Tear production was recorded weekly by using phenol red
 threads (PRT) (Zone Quick Tread, Oasis Medical, San Dimas, CA, USA) placed
 into the lateral canthus of the eye for 20 s, and the amount of tear fluid
 was recorded in mm.  Clinical scores (0-4) of eyelid inflammation, corneal
 opacity, and corneal fluorescein retention were recorded, following slit
 lamp biomicroscopy (KOWA SL-17, Torrance, CA), three times per week
 through day 44 after HSCT with the observer masked to the treatment
 groups. The most prominent clinical signs affecting
 ocular tissue following allogenic HSCT manifested as lid margin, cornea,
 and corneal fluorescein retention score changes. Individual mice received
 a score of 0-4 for the grade of each category.    Each
 database sheet (Tab) has the same organizational format, with columns of
 Mouse ID (identification), Cohort, Treatment (CSA-cyclosporine;
 AAV-HLAG1/5 (subconjunctival) or no treatment. The timepoint is the day
 following HSCT. and finally, the score assigned (based on the description
 previously).  Eyelid Score Tab: Lid margins were
 evaluated on the severity of edema, skin swelling, and if the eyelids were
 able to open. Cornea Scores Tab: The clinical scale of
 corneal involvement ranged from normal and clear to variable stages of
 keratopathy to corneal ulcerations. Fluorescein
 Retention Scores Tab: Corneal fluorescein retention scores were evaluated
 based on geographic coverage and density of the stain.
 PRT Scores Tab: Phenol red thread tear evaluation.  This a
 measurement of tear production using the PRT thread.  The values are read
 in mm of wetting over 20 seconds.  Weights % Tab: body
 weights and percentage of baseline of the mice over time
 Histology Scores Tab: Histology scores: Additionally, histologic
 images of the cornea and limbus were collected and analyzed using ImageJ
 software to measure epithelial, stromal, and total corneal thickness.  
 Epithelial thickness is the thickness in uM of the corneal epithelium,
 corneal thickness is the total corneal thickness in uM, and ratio is the
 ratio of the epithelial thickness to total corneal thickness.  The #cells
 at the limbus is the number of mononuclear cells at the superior limbus
 quantitated in an area measuring 20x20um from the limbus extending
 posteriorly.  
description: Microscoft Excel database