EAU was induced as described previously (Luger et al., 2008 (link)), with 150 µg IRBP emulsified in an equal volume of CFA containing 2.5 mg/ml Mycobacterium tuberculosis strain 37RA (Sigma-Aldrich) and 0.5 µg of Bordetella pertussis toxin. Some mice were treated with 0.5 mg of anti–IL-27 antibody (provided by J. Van Snick and C. Uyttenhoeve, Ludwig Institute for Cancer Research, Brussels, Belgium) or with isotype control antibody on days 0, 7, and 14. For depletion of NK cells and CD8+ cells, 0.2 mg of anti-NK1.1 antibody (PK136; Bioxcell) or 1 mg of anti-CD8 antibody (YTS169; Harlan), respectively, (or corresponding isotype controls) were injected on day 0, 3, and 6. Disease was evaluated by fundus examination on a scale of 0–4 based on the number, type, and size of lesions and extent of inflammation (Agarwal et al., 2012 (link)).
Protocol detail
Experimental Autoimmune Uveitis Induction and Treatment
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Anti antibody
Bordetella pertussis
Cancer
Cd8 cells
Il 27
Inflammation
Isotype
Mice
Mycobacterium tuberculosis
Nk cells
Strain
Toxin 5
Corresponding Organization :
Other organizations : National Institute of Allergy and Infectious Diseases, National Institutes of Health, National Eye Institute, Sun Yat-sen University
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Variable analysis
independent variables
- Anti-IL-27 antibody treatment (0.5 mg on days 0, 7, and 14)
- Anti-NK1.1 antibody treatment (0.2 mg on days 0, 3, and 6) for NK cell depletion
- Anti-CD8 antibody treatment (1 mg on days 0, 3, and 6) for CD8+ cell depletion
- Disease severity evaluated by fundus examination on a scale of 0-4 based on the number, type, and size of lesions and extent of inflammation
- Induction of EAU by IRBP emulsified in an equal volume of CFA containing 2.5 mg/ml Mycobacterium tuberculosis strain 37RA and 0.5 µg of Bordetella pertussis toxin
- Isotype control antibody treatment
- Positive control: Induction of EAU by IRBP emulsified in an equal volume of CFA containing 2.5 mg/ml Mycobacterium tuberculosis strain 37RA and 0.5 µg of Bordetella pertussis toxin
- Negative control: Isotype control antibody treatment
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