Citations

Mice, PCR, and EAE immunization:

To induce EAE, adult female mice were injected subcutaneously in the flank and at the tail base with 200µg of myelin oligodendrocyte glycoprotein (MOG) 3555 peptide emulsified in complete Freunds adjuvant (BD Biosciences, San Jose, CA, USA) supplemented with 600 g of Mycobacterium tuberculosis (strain H37Ra; BD Biosciences). Two i.p. injections of 400 ng pertussis toxin (List Biological Laboratories, Denver, CO, USA) were given 0 and 48 h later. …

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)

… Mice also received intraperitoneal injections of 200ng pertussis toxin (List Biological Laboratories) in sterile phosphate-buffered saline (PBS) immediately after MOG administration and 48h later. CFA control mice were similarly induced but lacked MOG …

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 was induced in female mice (8 to 12 weeks) by subcutaneous injection of MOG3555 peptide (100g, CSBio) emulsified in complete Freunds adjuvant. Pertussis toxin (250ng, List Biologicals) was administered intraperitoneally on the day of and one day after MOG 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)

… Directly and 48 hours after immunization, mice were intraperitoneally injected with 200 ng pertussis toxin (List biological laboratories, CA) (51). …

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)

… The mice also received ip injection of pertussis toxin (300 ng in 100 µl PBS; List Biological Lab, USA) at the time of immunization and again 48 hours later. …

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)

… and Pertussis toxin was purchased from List Biological Laboratories (Campbell, CA) …

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)

Bladder cancer permanent cell lines:

Experiments were carried out with commercially available permanent human CAL-29 tumour cell line (TCC), established from the primary lesion of a fatally invasive, metastatic TCC of the bladder (grade IV, stage T2) from an 80-year-old woman before treatment. …

Normal human urothelial cells were grown to confluence in complete keratinocyte serum-free medium (cKSFM) with human recombinant epithelial growth factor (5ng/mL) bovine, pituitary extract (50mg/mL; Gibco), supplemented with cholera toxin (30ng/mL; List Biological Laboratories, Campbell, CA, USA) under standard conditions.

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

Reagents:

… CTB (choleragenoid, 103B) and Clostridium difficle toxin B (TcdB) (6246-GT-020) were purchased from List Biological Laboratories and R&D Systems, respectively. …

Measurement of IL-1 production in vitro:

For measuring IL-1 production in vitro, 2 10⁵ cells per well in 96-well plates were first cultured for 5 h in the absence or presence of LPS O55:B5 or LPS O111:B4. Then CTB, ATP, R848 or TcdB was added at indicated concentrations and further cultured for indicated hours. A pan-caspase inhibitor, Z-VAD-FMK (4800-520, MBL), and a protein kinase A (PKA) inhibitor, H-89 (10010556, Cayman Chemical), were added 3 h before addition of CTB or ATP. Dimethyl sulfoxide (DMSO, Nacalai Tesque) was used as a solvent control for inhibitors. …

Measurement of IL-1 production in vivo:

Mice were injected intra-peritoneally (i.p.) with 100 g LPS O55:B5 per head. Five hours later, CTB was injected i.p. at 100 µg per head. PBS was used as a control for CTB. Then further 6 h later, peritoneal lavage fluids were harvested and subjected to ELISA for IL-1.

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

Assessment of toxin neutralizing antibodies

Toxin neutralization assay was performed as previously described^4,5,6. Briefly, sample dilutions were added to J774 macrophages cultured in cultured in RPMI supplemented with 10% fetal calf serum. Bacillus anthracis lethal toxin (LeTx) [i.e., PA plus Bacillus anthracis lethal factor (LF, List Biological, Campbell, CA)] was then added to the plates. After overnight incubation, MTT (3-(4,5-dimethylthiazol-2-yl)2,5-diphenyl tetrazolium bromide; Sigma-Aldrich) was added to assess the viability of macrophages as a function of redox potential. Toxin neutralizing antibody titers were determined as the lowest concentrations of sera  that protect macrophages from the cytotoxicity of LeTx.

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).

… Pertussin Toxin List Biological Laboratory Cat. #181 …

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