Citations

2.2. Induction of EAE

rMOG (2mg/ml) was emulsified with an equal volume of complete Freund’s adjuvant (CFA, 4mg/mL, Sigma, F5506). EAE animal model was induced by subcutaneous injection of the above emulsion into the back. 300ng pertussis toxin (List Biological Laboratories, 180228A1) was injected into the tail vein on the day of immunization and the second day after immunization. Female mice under the same conditions were used as control, and CFA was injected subcutaneously. …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… For CT-B processing, sections were blocked in 10% horse serum (2h, room temperature) and incubated in anti-CT-B made in goat (List Biological #703; 1:4000 dilution) in TBS-TX/3% horse 234 serum for 2-3 days at 4oC. After thorough washing, sections were further incubated in ImmPRES …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Anti-Cholera Toxin B Subunit (Goat) List Biological Laboratories Cat#703; …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

4.4. EAE Induction and Clinical Score Assessment

Active EAE was induced as previously described [34]. Briefly, mice were subcutaneously injected with 200 μg of MOG35–55 peptide (Genscript, RP10245) emulsified in incomplete Freud’s adjuvant (Sigma, F5506; St. Louis, MO, USA) with mycobacterium tuberculosis H37Ra (BD Difco, BD 231141; Pittsburgh, PA, USA) and intraperitoneally injected with 200 ng of Pertussis Toxin (List Biological Laboratories, Campbell, CA, USA). …

Author did not specify which List Labs Pertussis Toxin was utilized. List Labs provides the following Pertussis Toxin products:
• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)
• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Pertussis toxin (PTX, cat: 181) was purchased from List Biological Laboratories (CA, USA). …

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

2.2. Experimental design

… The two experimental sessions were held between two and four weeks apart to mitigate potential carry-over effects from LPS-mediated immune activation. …

The LPS infusion was prepared on-site immediately before administration, following manufacturer protocols for reconstitution and dilution under sterile conditions. In short, the product was reconstituted and initially diluted with sterile water, then diluted to the final dose in 30 mL sterile saline within 15 min of administration. For each participant, a single fresh vial of Clinical Center Reference lyophilized endotoxin (lipopolysaccharide/LPS; lot 94332B1) was used, prepared from Escherichia coli O113:H10:K by List Biological Laboratories, Inc. The infusion was administered at a slow rate of 1 mL/min over 30 min using an infusion pump (Taudorf et al., 2007). In the first session, LPS was administered at a very low dosage of 0.1 ng/kg of participant body weight, and in the second session at a low dosage of 0.4 ng/kg. …

• Product #9433 – GMP LPS Lipopolysaccharide List™ Hpt™ From Escherichia Coli Type O113

smFISH and immunostaining

… For immunostaining, sections were permeabilized in PBS + 0.3% Triton X-100, blocked for 1 h in block buffer (PBS + 0.3% Triton + 10% normal donkey serum) and incubated with primary antibodies in block buffer at 4 °C overnight. Slides were washed three times, incubated in secondary antibodies in block buffer for 1 h at room temperature and washed three times, and a coverslip was applied with ProLong Gold Antifade Reagent. Primary antibodies included: chicken anti-GFP (Aves Labs, GFP-1010, 1:1,000), rabbit anti-c-Fos (Synaptic Systems, 226 003, 1:5,000), goat anti-ChAT (Millipore, AB144P, 1:100) and goat anti-CTB (List Labs, 703, 1:1,000). Species-specific donkey secondary antibodies conjugated to Alexa Fluor 488, 568 or 647 were obtained from Life Technologies or Jackson ImmunoResearch and used at a 1:500 dilution. …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)