EAE was induced as described16 (link),25 (link). All mice used were on the C57Bl/6J background. EAE was induced using 150 μg of MOG35–55 (Genemed Synthesis, 110582) mixed with freshly prepared complete Freund’s Adjuvant (using 20 ml Incomplete Freund’s Adjuvant (BD Biosciences, BD263910) mixed with 100 mg Myobacterium tuberculosis H-37Ra (BD Biosciences, 231141)) at a ratio of 1:1 (v/v at a concentration of 5 mg ml−1). All mice received 2 subcutaneous injections of 100 μl each of the MOG/CFA mix. All mice then received a single intraperitoneal injection of pertussis toxin (List Biological Laboratories, 180) at a concentration of 2 ng μl−1 in 200 μl of PBS. Mice received a second pertussis toxin injection at the same concentration two days after the initial EAE induction. Mice were monitored and scored daily thereafter. EAE clinical scores were defined as follows: 0, no signs; 1, fully limp tail; 2, hindlimb weakness; 3, hindlimb paralysis; 4, forelimb paralysis; 5, moribund; as described previously16 (link),20 (link),22 (link),25 (link),54 (link). Mice were randomly assigned to treatment groups. All mice were scored blind to genotype.
Myobacterium tuberculosis h37ra
Manufactured by BD
Mycobacterium tuberculosis H37Ra is a laboratory strain of the Mycobacterium tuberculosis bacterium. It is a non-virulent strain commonly used in research and testing applications.
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4 protocols using myobacterium tuberculosis h37ra
1
Induction and Evaluation of Experimental Autoimmune Encephalomyelitis
2
Intranasal Delivery of PTN for EAE in Mice
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EAE was induced in 8-12 week female C57Bl/6J mice using 150 μg of MOG35–55 (Genemed Synthesis, 110582) mixed with freshly prepared complete Freund’s Adjuvant [using 20 ml Incomplete Freund’s Adjuvant (BD Biosciences, #BD263910) mixed with 100 mg Myobacterium tuberculosis H37Ra (BD Biosciences, #231141)] at a ratio of 1:1 (v/v at a concentration of 5 mg ml−1). All mice received 2 subcutaneous injections of 100 μl each of the MOG35-55/CFA mix. All mice then received a single intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories, #180) at a concentration in 200 μl of PBS. Mice received a second pertussis toxin injection at the same concentration two days after EAE induction. Mice were monitored and scored daily thereafter. EAE clinical scores were defined as follows: 0, no signs; 1, fully limp tail; 2, hindlimb weakness; 3, hindlimb paralysis; 4, forelimb paralysis; 5, moribund.
Intranasal delivery of PTN or vehicle (PBS) was started on day 16 after EAE induction. 20 µl of vehicle or 300µg/kg PTN (solved in PBS) was applied drop by drop on nostrils.
Intranasal delivery of PTN or vehicle (PBS) was started on day 16 after EAE induction. 20 µl of vehicle or 300µg/kg PTN (solved in PBS) was applied drop by drop on nostrils.
3
Induction of Experimental Autoimmune Encephalomyelitis
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EAE was induced in 8-12-week female C57Bl/6J/NOD/ShiLtJ mice using 150 μg of MOG35–55 (Genemed Synthesis, 110582) mixed with freshly prepared complete Freund’s Adjuvant (using 20 ml Incomplete Freund’s Adjuvant (BD Biosciences, #BD263910) mixed with 100 mg Myobacterium tuberculosis H37Ra (BD Biosciences, #231141)) at a ratio of 1:1 (v/v at a concentration of 5 mg/ml). All mice received two subcutaneous injections of 100 μl each of the MOG35-55/CFA mix. All mice then received a single intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories, #180) at a concentration in 200 μl of PBS. Mice received a second pertussis toxin injection at the same concentration two days after EAE induction. Mice were monitored and scored daily thereafter. EAE clinical scores were defined as follows: 0, no signs; 1, fully limp tail; 2, hindlimb weakness; 3, hindlimb paralysis; 4, forelimb paralysis; 5, moribund.
4
Induction of Experimental Autoimmune Encephalomyelitis in Mice
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EAE was induced as previously described52 (link). In brief, 8–12-week-old female C57Bl/6J mice were immunized using 150 μg of MOG35–55 (Genemed Synthesis, 110582) mixed with freshly prepared Complete Freund’s Adjuvant (using 20 ml of InComplete Freund’s Adjuvant (BD Biosciences, no. BD263910) mixed with 100 mg of Myobacterium tuberculosis H37Ra (BD Biosciences, no. 231141)) at a ratio of 1:1 (v/v at a concentration of 5 mg ml−1). All mice received two subcutaneous injections of 100 μl each of the MOG35-55/CFA mix. All mice then received a single intraperitoneal injection of 200 ng of pertussis toxin (List Biological Laboratories, no. 180) at a concentration in 200 μl of PBS. Mice received a second pertussis toxin injection at the same concentration 2 d after EAE induction. Mice were monitored and scored daily thereafter. EAE clinical scores were defined as follows: 0, no signs; 1, fully limp tail; 2, hindlimb weakness; 3, hindlimb paralysis; 4, forelimb paralysis; 5, moribund.
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