Citations

Disruption of BBB and passive transfer of IgG from NMOSD patients

Animal procedures are summarized in Fig. 1a. To disrupt the BBB, mice were anaesthetized with intraperitoneal (i.p.) injection of ketamine (100 mg/kg) and xylazine (10 mg/kg). They received subcutaneous injections of complete Freund’s adjuvant (CFA, BD Biosciences, USA) containing heat-killed H37Ra Mycobacterium tuberculosis (Difco, USA) at 4 sites (50 μg in 50 μl CFA each site) on the hind flank on 7 days before IgG transfer. In addition, mice received i.p. injections of pertussis toxin (PTx, 200 ng in 0.2 ml PBS, List Biological Laboratories, USA) on 7 and 3 days prior to IgG transfer.

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)

EAE Induction and Scoring

… Two intraperitoneal injections of pertussis toxin, Bordatella pertussis, (List Biological Labs) were also administered to all mice on the day of induction and 48 hours thereafter. …

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)

Active EAE

… Immunization was administered subcutaneously (s.c.) on the back and tail base. Pertussis toxin (250 ng per mouse; List, Biological Laboratories, Campbell, CA, USA) in 200µl PBS was injected intraperitoneally twice, on the day of immunization (day 0) and again 2 days later (day 2). …

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)

Reagents

Lethal factor (LF) was purchased from List Biological Laboratories (#169, recombinant from B. anthracis)

• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Protein expression and purification

Upon addition of double molar excess of trypsin inhibitor (Sigma Aldrich), PA7 was further purified by size exclusion chromatography using a Superdex 200 column (GE Healthcare). Lyophilized LF (List Biological Lab. Inc., Lot#1692A1B) were resuspended in water according to the manufacturer’s manual and mixed with PA7 in a molar ration of 10:1. …

• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

Chemicals for in vitro experiments

The following products were purchased from the indicated commercial sources: … Shiga-like toxin 2 (Stx, List Biological Laboratories, Inc.), …

• Product #162L – Shiga Toxin 2 from Escherichia coli

BMDM isolation and culture.

… TcdB toxin (PYRIN inducer, 0.05µg/mL, List Biological Laboratories) or flagellin (NLRC4 trigger, 1g/mL, Invivogen) were used to stimulate various canonical inflammasomes …

• Product #155 – Toxin B from Clostridium difficile

… Immunofluorescence 1×105 SCC-4 cells/well (biological triplicates) control and Clostridium difficile Toxin A – ToxA (List Biological Labs, Campbell, CA, USA) treated: 1, 2 and 4 µg/mL cultured on three-dimensional MatrigelTM for 24 h. …

• Product #152C – Toxin A from Clostridium difficile

Botulinum neurotoxin and its administration

Research-grade, purified, native BoNT serotype A1 (BoNT-A; 150 kDa) was purchased from List Biological Laboratories (Campbell, CA, USA). It was received as a lyophilized powder, which was reconstituted in 1 mg/mL phosphate buffer saline/bovine serum albumin (0.1%) to obtain a stock solution (0.1 mg/mL), which was aliquoted and kept frozen at −80 °C. Dilutions of aliquots were performed with 0.2% GPB to obtain the final concentrations (see below). …

… Adjuvanted formulations included 2µg of cholera toxin (List Biological Laboratories, 101B), 10µg of CTB (List Biological Laboratories, 103B), or …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt