For TFH and TFR cell studies mice were immunized with 100μg MOG 35–55 (UCLA biopolymer facility), MOG 1–125 or NP18-OVA (Biosearch Technologies) emulsified 1:1 with H37RA Mtb. CFA (Difco laboratories) in the flanks as previously described(25 (link)). 7 days later draining lymph node or blood was harvested for analysis. For studies of TFH and TFR cell differentiation in PD-L1 conditional knockout mice, all mice received 200μg anti-PD-L2 (clone 3.2) on d2 and d4. For EAE induction, similar emulsions were prepared as above but contained an extra 2mg/ml H37RA Mtb. Mice were additionally given 200ng pertussis toxin i.p. on days 0 and 2. Clinical disease was assessed using the following criteria: 1 = limp tail, 2 = weak gait, 3 = hind limb paralysis, 4 = hind+forelimb paralysis, 5 =moribund as described previously(26 (link)).
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Induction and Evaluation of Autoimmune Encephalomyelitis
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Corresponding Organization : Brigham and Women's Hospital
Other organizations : Stanford University, Broad Institute, Dana-Farber Cancer Institute
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Variable analysis
independent variables
- Immunization with 100μg MOG 35–55, MOG 1–125 or NP18-OVA emulsified 1:1 with H37RA Mtb and CFA
- Administration of 200μg anti-PD-L2 (clone 3.2) on d2 and d4
- Induction of EAE with emulsion containing extra 2mg/ml H37RA Mtb and 200ng pertussis toxin i.p. on days 0 and 2
- Draining lymph node or blood harvested for analysis 7 days after immunization
- Clinical disease assessed using a 5-point scale (1 = limp tail, 2 = weak gait, 3 = hind limb paralysis, 4 = hind+forelimb paralysis, 5 = moribund)
- Mice were immunized as previously described(25)
- Positive control: None specified
- Negative control: None specified
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