The reagents used in this study were purchased as described: THC and CBD from Cayman Chemical (Michigan, USA), myelin oligodendrocyte glycoprotein (MOG35−55) peptide H-MEVGWYRSPFSRVVHLYRNGK-OH (PolyPeptide Laboratories, San Diego, CA, USA). Mycobacterium tuberculosis (strain H37Ra) (BD, Franklin Lakes, NJ, USA), complete Freund's adjuvant (Fisher, Hampton, NH, USA), Pertussis toxin (List Biological Laboratories, Campbell, CA, USA), Percoll, GE Healthcare Life Sciences (Pittsburgh, PA, USA); Neural Tissue Dissociation Kit (P) (Miltenyi Biotech, Auburn, CA, USA), RBC lysis buffer (Sigma-Aldrich, St. Louis, MO, USA), RPMI 1640, l-glutamine, HEPES, phosphate-buffered saline, and fetal bovine serum (VWR, West Chester, PA, USA), ELISA Max Kits IL-10, IL-17A, IFN-γ, IL-6, IL-1β, TNF-α, and TGF-β and FITC Annexin V/-PI apoptosis kit (Biolegend, San Diego, CA). EasySep PE selection kit (Stemcell Technologies, Cambridge, MA, USA), Propidium Iodide (PI)/RNase Staining Solution (Cell Signaling Technology, Danvers, MA, USA), miRNeasy Mini Kit, miScript II RT Kit and miRNAs primers (Qiagen, Valencia, CA), mRNAs primers (Integrated DNA technologies, Coralville, IA, USA) and SsoAdvanced™ Universal SYBR® Green Supermix (Bio-Rad, Hercules, CA, USA).
Mycobacterium tuberculosis strain h37ra
Manufactured by BD
Sourced in United States, Italy, Canada
Mycobacterium tuberculosis (strain H37Ra) is a laboratory strain of the bacterium Mycobacterium tuberculosis, the causative agent of tuberculosis. This strain is commonly used in research and testing applications.
Automatically generated - may contain errors
Lab products found in correlation
Pertussis toxin, by List Biological Laboratories (5 mentions)
Pertussis toxin, by Merck Group (4 mentions)
Complete freund adjuvant (cfa), by BD (3 mentions)
Incomplete freund s adjuvant, by Merck Group (3 mentions)
Complete freund s adjuvant, by BD (3 mentions)
Mog35 55, by Biomatik (2 mentions)
Pertussis toxin, by Alexis Biochemicals (2 mentions)
Ssoadvanced universal sybr green supermix, by Bio-Rad (1 mentions)
Complete freund s adjuvant, by Thermo Fisher Scientific (1 mentions)
Percoll, by GE Healthcare (1 mentions)
Phosphate buffered saline (pbs), by Avantor (1 mentions)
Fetal bovine serum (fbs), by Avantor (1 mentions)
L glutamine, by Avantor (1 mentions)
Hepes, by Avantor (1 mentions)
Rbc lysis buffer, by Merck Group (1 mentions)
Propidium iodide rnase staining solution, by Cell Signaling Technology (1 mentions)
Mirneasy mini kit, by Qiagen (1 mentions)
Rpmi 1640, by Avantor (1 mentions)
Neural tissue dissociation kit p, by Miltenyi Biotec (1 mentions)
Easysep pe selection kit, by STEMCELL (1 mentions)
28 protocols using mycobacterium tuberculosis strain h37ra
1
Experimental Protocol for Mouse EAE Model
2
Experimental Autoimmune Encephalomyelitis Induction
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Preparations of 200-μg MOG35–55 peptide emulsified in CFA (no. 263910, BD Biosciences, San Jose, CA) supplemented with 200 μg of Mycobacterium tuberculosis (strain H37Ra, BD Biosciences) were injected subcutaneously into the lower flanks of 8-week-old female C57BL/6J mice (Jackson Laboratory, Bar Harbor, MN). Two intraperitoneal (i.p.) injections of 400-ng pertussis toxin each (no. 181, List Biological Laboratories, Denver, CO) were administered 0 and 48 h later. Mice were given i.p. injections of guanabenz or vehicle (sterile 0.9% NaCl) daily beginning PID7. Treatment groups were randomized by assignment before injections on the basis of cage number. Mice were monitored for clinical symptoms beginning PID7 and scored daily (0=healthy, 1=flaccid tail, 2=ataxia and/or paresis of hindlimbs, 3=paralysis of hindlimbs and/or paresis of forelimbs, 4=tetraparalysis, 5=moribund or death). To minimize daily scoring bias, mice were blindly assessed for clinical score before confirmation of treatment dosage and injection. Treatments were administered at the same time every day, ±1–2 h. Data from all mice were included in the study, as per previously established criteria.
3
Inducing Experimental Autoimmune Uveitis in Mice
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To induce EAU, B10RIII mice were injected subcutaneously with 100 μL of 20 μg R161 peptide, suspended in phosphate-buffered saline (PBS) and emulsified in Freund’s complete adjuvant (CFA, 1:1 v/v, BD Pharmingen, San Diego, CA, USA), containing Mycobacterium tuberculosis strain H37RA (2.5 mg/mL, BD Pharmingen). The mice were scored by fundoscopy and histologic examination in accordance with the previously described protocols [12 (link)].
4
MOG-Induced Experimental Autoimmune Encephalomyelitis
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In the EAE induction, mice were immunized with MOG35–55 peptide and euthanized 14 d later. To this end, 200 μg of MOG35–55 peptide (GL Biochem Ltd.) was emulsified in incomplete Freund’s adjuvant containing 0.5 mg/ml Mycobacterium tuberculosis (strain H37Ra; BD Diagnostics), and 200 μl of the mixture was injected subcutaneously to the mice. Pertussis toxin was not administered as part of the EAE induction.
5
Experimental Autoimmune Encephalomyelitis Induction and Treatment
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EAE was induced in mice (8–12 weeks of age) as previously described21 (link). Briefly, mice were injected subcutaneously with 200 µL of recombinant MOG35–55 (Biomatik Corporation), emulsified in incomplete Freund’s adjuvant (Sigma, Oakville, Canada) and supplemented with 4 mg/mL Mycobacterium tuberculosis (strain H37Ra, BD Biosciences, Mississauga, Canada) at four different sites with 50 µL per site. 200 ng of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA) was injected intraperitoneally (i.p.) on day 0 and 2 after immunization. Mice were assessed for clinical signs of disease at the beginning of immunization. 3 nmol/g of GluA2-G-Gpep or TAT-control peptide was subsequently injected (i.p.) daily starting from day 10 of immunization for 18 days until day 28. Our group had previously shown that this peptide concentration effectively disrupted the GluA2-GAPDH interaction21 (link).
6
Induction of Experimental Autoimmune Encephalomyelitis in Mice
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Female C57BL/6 mice (8–12 weeks of age) were purchased from Charles River Laboratories. Mice were injected subcutaneously (s.c.) in the back with 200 μL of recombinant MOG35-55 (200 μg, Biomatik Corporation, Cambridge, ON, Canada), which was emulsified in incomplete Freund's adjuvant (Sigma, Oakville, ON, Canada) and supplemented with 4 mg/mL Mycobacterium tuberculosis (strain H37Ra, BD Biosciences, Missisauga, ON, Canada) in four different sites (50 μL per site). Two hundred nanograms of pertussis toxin (List Biological Laboratories, Inc., Campbell, CA, USA) was injected intraperitoneally (i.p.) on days 0 and 2 after immunization. Mice were assessed daily for clinical signs of disease starting on the day of immunization. Clinical scoring was performed according to the following criteria: 0, asymptomatic; 0.5, distal paresis of the tail; 1, complete tail paralysis; 1.5, paresis of the tail and mild hind limb paresis; 2, unilateral severe hind limb paresis; 2.5, bilateral severe hind limb paresis; 3, complete bilateral hind limb paralysis; 3.5 complete bilateral hind limb paralysis and paresis of one front limb; 4, complete paralysis (tetraplegia). All animal procedures used were in accordance with the approved Centre for Addiction and Mental Health animal research ethics committee protocol.
7
Evaluating Bile Acid's Role in Autoimmune Uveitis
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TGR5-/- mice were obtained from Viewsolid Biotech (Beijing, China). TGR5+/+ (C57BL/6J) mice were purchased from Jackson Laboratory (Bar Harbor, ME, USA). EAU was induced in 6- to 8-week-old female mice through subcutaneous injection of 300 µg human IRBP651-670 dissolved in 0.2 ml emulsion 1:1 (vol/vol) supplemented with CFA containing 2.5 mg/mL Mycobacterium tuberculosis strain H37Ra (BD Biosciences, USA). The mice also received intraperitoneal injections of 0.5 µg Bordetella pertussis toxin (Sigma-Aldrich, MO, USA) suspended in 0.1 ml PBS. The toxin was injected once on the first day and after 2 days. Then, EAU mice were fed a special LCA diet (0.01%) or normal diet. Clinical manifestations and histopathological changes in the mice were evaluated as previously described 1 (link).
8
Guanabenz Reduces Relapse Severity in Remitting-Relapsing MS Mouse Model
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Example 6
guanabenz reduces the severity of symptom relapse in an in vivo mouse model of remitting/relapsing MS (
9
Induction of Experimental Autoimmune Encephalomyelitis
10
Immunization Protocol for Mouse GH
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Purified recombinant mouse growth hormone (mGH, 5 mg/mL) was emulsified 1-to-1 in CFA (which contained 5 mg/mL of heat-killed Mycobacterium tuberculosis, strain H37Ra, from BD Diagnostic Systems, Sparks, MD, U. S. A.), and injected subcutaneously on day 0 in a volume of 100 μl (50 μL in the dorsal hind leg region and 50 μL in the contralateral inguinal region). Protein emulsions were injected again on day 7 in the opposite sites. Mice were sacrificed at 28 (early stage) or 56 (mid-late stage) days post-immunization.
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