Bacterial Toxin Research Citations

Stimulations and viral infections

N/TERT1 cells were plated at 5 × 104 cells/96-well plate for cytokine release experiments and at 2.5 × 105, 5 × 105, or at 1 × 106 cells per 24-, 12-, or 6-well plate, respectively. The next day, the cells were treated as indicated with 2 µM Val-boroPro (ApexBio and Biozol), 2 µM ANS (Biomol), 100 ng/mL TNFα (PeproTech), 6.5 µM Nigericin (Sigma Aldrich), 0.1 µM B/B homodimerizer (AP20187, MedChem Express), 20 µM Z-VAD-FMK (Peptanova), 10 µM cytarabine (Ara C, Sigma Aldrich), 1 µM MLN4924 (MLN), 1 µg/mL anthrax lethal factor, and 1 µg/mL protective antigen (both List Biological Labs). …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

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

Immunogenicity Studies of PA83 variants in Mice

Immunogenicity studies were performed with 3 and 10 μg of plant-produced dPA83(E), dPA83(P) and commercial recombinant PA (rPA, List Biological Laboratories, Campbell, CA, used as a positive control) proteins, administered intraperitoneally in seven- to eight-week-old Balb/c mice (6 animals/group). Mice were injected with three doses of protein samples adsorbed in 0.3% Alhydrogel at three-week intervals (day 0, 21 and 42). …

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

… Anthrax lethal toxin List Biological Laboratories Inc. #171B/#172B …

Treatments

See Table S1 for list of Figures/panels in each figure and corresponding dose and exposure time. General information for the methods is provided here. To induce cytokine release from macrophages, microglia and BMDM cells were primed for 4–5 h (or time indicated in Table S1)with 1 μg/mL of LPS in culture medium, and some cells were pre-treated with 1 μM of CP456773/MCC950 for 30 min before ATP stimulation. Cells were then pulsed with 5 mM ATP for 30 min. For cytokine induction by anthrax lethal toxin (LeTx), LPS-primed cells were treated with LeTx at a final concentration of 250 ng/mL (LF and PA) for 3 h. Cell supernatants were collected and frozen at −80°C until assayed.

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

… goat anti-EF (773L, List Labs, Campbell, CA, USA), …

2.4.4 Purification of PA and LF

PA83 was expressed and purified by the Barth group. After purification, PA83 was activated in vitro to PA63 and stored at -80 °C.
LF was purchased from List Biological Laboratories (see 2.1.7) and stored at -80 °C.

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… For the activation of NLRP3, nigericin was used at a final concentration of 5 µM. PA (List Biological Laboratories) and LFn-PrgJ (expressed and isolated at Nodthera) were added to yield final concentrations of 1 and 4 …

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

Animal challenge

Mice were randomly assigned to the control or experimental groups. Mouse TNF (CF09, Novoprotein, China)
and/or LT (LF (172C, List Biological Labs, USA) and PA (171E, List Biological Labs)) diluted in 200 μL of phosphate buffered saline (PBS) was intravenously injected into the mouse tail-vein with or without inhibitors of caspase (Z-VAD-FMK, 627610, Calbiochem, Germany) or p38α (TAK-715, HY-10456, MedChemExpress, USA) diluted at indicated doses. Mouse survival was monitored and recorded at indicated time points.

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Inflammasome activation

Late-log cultures of Salmonella enterica serovar Typhimurium strain SL1344 (a kind gift from Dr. Brad T. Cookson, University of Washington) were grown in L-broth containing 0.3 M NaCl with shaking at 37 °C. Bacteria were washed, resuspended in PBS, and added at a multiplicity of infection of 10 bacteria per macrophage. For activation of NLRP1b, BMM from BALB/c mice were exposed to 1 µg/mL anthrax lethal toxin, prepared as a 1:1 mix of protective antigen and lethal factor (List Biological). …

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Anthrax Protective Antigen List Biological Laboratories Cat# 171E …

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