Citations

2.3. EAE induction and NPC transplantation

Females 6 to 8 weeks old SJL mice were immunized with 175 μg Proteolipid protein (PLP) 139‐151 peptide diluted in normal saline (0.9% NaCl) and emulsified with complete Freund’s adjuvant containing 5 mg/ml H37RA (Difco Laboratories, Detroit, MI), supplemented with 4 mg/mL of heat-killed Mycobacterium tuberculosis (H37Ra, Difco, Lawrence, KS). Bordetella pertussis toxin (50ug, List Biological Industries) was diluted in 0.2 ml normal saline and injected intra-peritoneally immediately after subcutaneous injection of PLP emulsion. A second bolus of pertussis toxin was administered intra-peritoneally 48 h after primary injection of PLP emulsion. …

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)

2.2 BoNT

rBoNT/A1 gene synthesis, expression in E. coli, and purification methods are described by Hooker et al.15 rBoNT/A1 activity in vitro was confirmed through cell-based assays (SNAP25 cleavage) using rat embryonic spinal cord neuronal (eSCN) cultures. rBoNT/A1 activity ex vivo was shown in the mouse phrenic nerve hemidiaphragm.15

Research-grade purified nBoNT/A1 (150 kDa) was purchased from List Biological Laboratories. The molecular activity was previously confirmed in glycine and glutamate release assays, in SNAP25 cleavage using either rat eSCN or spinal cord neuronal cultures, and in the mouse nerve phrenic hemidiaphragm assay.10, 15 nBoNT/A1 was reconstituted in 1 mg/ml bovine serum albumin (Sigma-Aldrich) in Dulbecco’s phosphate-buffered saline (Sigma) to obtain a 0.1 mg/ml stock solution that was stored at −80°C as single-use aliquots. rBoNT/A1 was stored at −80°C, as single-use aliquots, at 0.1 mg/ml in phosphate-buffered saline (KH2PO4 1 mM, Na2HPO4 3 mM, NaCl 155 mM, pH 7.4). Working solutions were prepared with gelatin phosphate buffer (GPB) to yield the final desired concentrations (see below). GPB was prepared using 0.2% v/v gelatin (Prionex®, Sigma), 0.7% w/v Na2HPO4 (Emsure®, Merck), and sterile water for irrigation (Ecotainer®, Aqua B. Braun). The final pH was adjusted to 6.5 using orthophosphoric acid (85%; Merck).

• Product #130 – Botulinum Neurotoxin Type A from Clostridium botulinum

… EAE was induced with 50 µg myelin oligodendrocyte glycoprotein (MOG35-55; Synpeptide, Shanghai, China) emulsified with 1 mg/ml complete Freund’s adjuvant (CFA; Sigma-Aldrich, Oakville, Ontario, Canada), and 200 or 400 ng of pertussis toxin (PTX; lot #180235A1A, CHO cell assay 0.03 ng/ml; List Biological Laboratories, Campbell, CA), as previously described (Segal et al., 2020) …

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

Pertussis Toxin (PT) was purchased from List Biological Laboratories (Campbell, CA USA)

Induction of EAE. EAE was induced as described (17). In brief, acclimatized, 9-week-old, female C57BL/6J mice were immunized subcutaneously in the flank with 100 μg MOG emulsified 1:1 in IFA supplemented with 4 μg/mL of M. tuberculosis (H37Ra; Difco Laboratories). Animals received 200 ng by i.p. injection of PT on days 0 and 2 postimmunization. R-EAE was induced in acclimatized 9-week-old female SJL/J mice by immunization subcutaneously with 100 μL (50 μg) of PLP emulsified 1:1 in IFA containing 4 μg/mL of M. tuberculosis. …

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)

… Pertussis toxin was from List Biologicals.

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)

Experimental autoimmune encephalomyelitis

EAE was induced in 8–12 weeks old female C57BL/6J WT and ACKR1–/- C57BL/6J mice as described before [59, 60]. Pertussis toxin (List Biological Laboratories, Campbell, US) (300 ng in 100 μl PBS/mouse) was injected intraperitoneally (i.p.) on day 0 (immunization day) and day 2 post-immunization. …

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)

… After a location was found from which a compound movement resembling the p-Synergy could be evoked with low currents, the stimulating pipette was removed from the brain, cleaned with deionized water, and filled with 1% cholera toxin (CT); List Biological Laboratories …

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Animal studies and treatments

… Mice were treated with 0.5µg DT or CRM197 (List Biological) i.p. on day 0 only or on days 0, 4, 7, and 10. For in vivo competition assays, mice were treated on day 0 with 0.125-0.5µg DT and/or CRM197. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #149A – Diphtheria Toxin CRM197, Mutant

… Anthrax Lethal Factor (LF-A) (#171E) and Anthrax Protective Antigen (PA) (#169L) were obtained from List Biological Laboratory (California, USA). …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… On the same day as immunization and 2 days later, mice were also injected IP with 300 ng of pertussis toxin (List Biological Labs, Campbell, California, 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)