EAU was induced as described previously (Silver et al., 2015 (link)) using 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 (PTx). Mice were given i.p. injections of PBS, IFN-α or IFN-β (2000 IU, PBL) daily for 10 days starting from day 14 at the onset of disease. Disease was evaluated by fundus examination as described previously (Chen et al., 2013a (link), 2013b (link)).
Mycobacterium tuberculosis strain 37ra
Manufactured by Merck Group
Sourced in United States
Mycobacterium tuberculosis strain 37RA is a laboratory strain of the bacterium that causes tuberculosis. It is commonly used in research and testing related to this infectious disease.
Automatically generated - may contain errors
5 protocols using mycobacterium tuberculosis strain 37ra
1
Experimental Autoimmune Uveitis Induction and Evaluation
2
Induced Autoimmune Uveitis Protocol
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EAU was induced as described previously [29 (link)]. IL-27Rα−/− and WT littermates were immunized subcutaneously with 200 μL emulsion of 200 μg human IRBP1–20 (GPTHLFQPSLVLDMAKVLLD, Hanhong, China) in an equal volume of complete Freund’s adjuvant (CFA, Sigma-Aldrich, St. Louis, MO, USA) containing 2.5 mg/mL Mycobacterium tuberculosis strain 37RA (Sigma-Aldrich, St. Louis, MO, USA). In addition, these mice also received 0.5 μg of Bordetella pertussis toxin (PTX, Sigma-Aldrich, St. Louis, MO, USA) intraperitoneally on the day of immunization and the next day of immunization.
3
Experimental Autoimmune Uveitis Induction and Treatment
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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)).
4
Experimental Autoimmune Uveitis Induction
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EAU was induced using 150 μg IRBP1-20 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 as described [18] (link). Clinical EAU severity was evaluated by retinal fluorescent imaging system (field of view 1.8 mm) (Phoenix Micron IV)[29 (link),30 (link)] and scored on a scale of 0-4 as follows [18 (link),31 ]: 0.5, mild vasculitis and focal lesions; 1, moderate vasculitis, focal and lineal lesions; 2, severe vasculitis and infiltrations, multiple chorioretinal lesions; 3, confluent lesions, retinal hemorrhages; and 4, retinal detachment, retinal atrophy.
5
Induction and Treatment of Experimental Autoimmune Uveitis in Mice
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To induce EAU, C57BL/6 mice were immunized with a 200 µL emulsion of 150 µg human IRBP1–20 (GPTHLFQPSLVLDMAKVLLD, Hanhong, China) in an equal volume of complete Freund’s adjuvant (CFA, Sigma-Aldrich, St. Louis, MO, USA) containing 2.5 mg/mL Mycobacterium tuberculosis (strain 37RA) (Sigma-Aldrich, St. Louis, MO, USA) and 0.5 µg of Bordetella pertussis toxin (PTX, Sigma-Aldrich, St. Louis, MO, USA), as described in a previous study [36 (link)].
For treatment, EAU mice were intravitreally injected with mouse-recombinant IL-24 (200 ng in 2 µL) (R&D Systems, Minneapolis, MN, USA) into the left eye at the onset of disease. As a negative control, 2 µL of PBS was injected into the right eye. Eyes were examined by funduscopy at regular intervals using the Micron-IV retinal imaging system for small animals (Phoenix Research Laboratories, Pleasanton, CA, USA) and scored on a scale of 0–4, as described in previous study [37 (link)]. Experiments were terminated on day 6–8 after disease onset for eye collection.
For treatment, EAU mice were intravitreally injected with mouse-recombinant IL-24 (200 ng in 2 µL) (R&D Systems, Minneapolis, MN, USA) into the left eye at the onset of disease. As a negative control, 2 µL of PBS was injected into the right eye. Eyes were examined by funduscopy at regular intervals using the Micron-IV retinal imaging system for small animals (Phoenix Research Laboratories, Pleasanton, CA, USA) and scored on a scale of 0–4, as described in previous study [37 (link)]. Experiments were terminated on day 6–8 after disease onset for eye collection.
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