Bacterial Toxin Research Citations

Active EAE Induction

MOG35–55 peptide with amino acid sequence MEVGWYRSPFSRVVHLYRNGK was purchased from Mimotopes (Mimotopes, Victoria, Australia) and resuspended in 0.09% saline solution at a concentration of 2 mg/ml. Fresh complete Freund’s adjuvant (CFA) was reconstituted with 4 mg/ml non-viable, desiccated Mycobacterium tuberculosis H37 Ra (Cat#231141, BD, MD, USA) in incomplete Freund’s adjuvant (Cat#F5506, Sigma, Millipore-Sigma, MO, USA). Pertussis toxin from Bordetella pertussis in glycerol (Cat#179B, List Biological Laboratories, CA, USA) was freshly diluted in PBS to 3 μg/ml. MOG35–55/CFA emulsion was freshly reconstituted by mixing equal volume of 2 mg/ml MOG35–55 with CFA and vortexed for 30 min at room temperature. For each mouse, a total of 200 μl MOG35–55/CFA emulsion was subcutaneously injected at flank and scruff sites; 100 μl of 3 μg/ml pertussis toxin was administered intraperitoneally 1 h and 24 h following MOG35–55/CFA emulsion. Clinical scoring was performed starting 7 days following active EAE induction, every day until the completion of the experiment. …

• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Vaccine preparation and immunization:

DPX-rPA and alum-rPA were provided by IMV Inc. (Halifax, Canada), but AVA was provided by the CDC. Test and vehicle/control article formulations were prepared fresh on the day of dosing in accordance with the method provided. DPX-rPA was prepared as previously described¹⁹ containing a poly I:C (Thermo Fisher) or Pam3CSK4 (EMC Microcollections) adjuvant and a 10:1 mixture of DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine):cholesterol (Lipoid GmBH, Germany) or S100:cholesterol (Lipoid GmBH), see Supplementary Table 1 for complete formulation details for each figure. The source of rPA was DRDC, List Biologicals, or Pfenex Inc.²⁶ …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

… Two hours and forty-eight hours after immunization, all mice received an intraperitoneal injection of … pertussis toxin (List Biological Lab …

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:

… 300ng/mouse Pertussis toxin (List Biological Laboratories) in PBS was injected intra peritoneal (ip) on days 0 and 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)

Acute lung inflammation model:

LPS (E. coli O55:B5 from List Biological Laboratories) (20 g) in 50 l of PBS was instilled directly into the tracheas of mice sedated with isofluorane (Baxter) using a modified feeding needle (59). Mice were sacrificed using i.p. injection of pentobarbital sodium solution.

• Product #203A – LPS from Escherichia coli O55:B5

… B. anthracis LF was purchased from List Biological Laboratories. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

EAE Induction:

… In addition, 500 ng pertussis toxin (PT; List Biological Laboratories, CA, USA) was injected by intraperitoneally on days 0 and 2 after 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)

Chemicals and Reagents:

Diphtheria toxin (DT) was obtained from List Biological Laboratories (Campbell, CA. Cat. No. 150), reconstituted in PBS and stored at 80C. Mice received a dose of DT (10ngg¹ in 0.1ml PBS)³⁰, injected intraperitoneal, every other day.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Reagents:

… B. anthracis lethal factor (LF) was acquired from List Biologicals (#172C). …

J774.A1 culture and stimulation:

J774.A1 cells were cultured in IMDM (Lonza) containing 10% (vol/vol) heat-inactivated FBS, 1% (vol/vol) nonessential amino acids (Lonza), 100 U/ml penicillin, and 100 mg/ml streptomycin at 37C in a humidified atmosphere containing 5% CO₂. Cells were scraped for passages and, for experiments, counted and seeded into 96-well plates in culturing medium. On the next day, cells were changed to fresh media and stimulated with vehicle or 1 g/ml PA combined with 0.5 g/ml LF (LeTx), VBP (0.1% TFA in DMSO) or 1G244 (DMSO) at a final concentration of 10 M or 25 M.

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

… To compare the potency of BoNT/D with BoNT/A (subtype BoNT/A1; List Laboratories Inc.), the drug substance of incobotulinumtoxinA, concentration-response curves were constructed from three data points, each determined at least in triplicate, fitted to the logarithmic functions y(BoNT/D) = -25.548Ln(x) + 233.4, R2 = 1.0 and y(BoNT/A) = -37.817Ln(x) + 259.88, R2 = 0.9885. …