Bacterial Toxin Research Citations

… Pertussis toxin (PTX, 0.4 µg/mouse/day; List Biological Laboratories Inc., Campbell, CA) in phosphate-buffered saline (PBS) was injected intraperitoneally at Day 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)

Reagents

… C. difficile TcdB toxoid (formaldehyde inactivated; List Biological Laboratories Inc) was used at 5 μg/mL.

… 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

… For the EAU model, the mice were subcutaneously injected with IRBP (500 µg) and intraperitoneally injected with 1 µg of pertussis toxin (List Biological Laboratories, Campbell, CA, 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)

… Mice were injected intraperitoneally with pertussis toxin (200 ng; List Biologicals) 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)

… Staphylococcus Enterotoxin B (SEB) was purchased from List Labs; …

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

Materials

… Botulinum neurotoxin type A toxoid from Clostridium botulinum (BoNT) and anti-botulinum neurotoxin type A (Chicken lgY) (Anti-BoNT) were obtained from List Labs, …

• Product #730 – Anti-Botulinum Neurotoxin, Type A (Chicken lgY)

Mouse studies

EAE was induced in C57Bl/6J mice (Jackson Laboratory) and SJL/J mice, aged 8–15 weeks. Complete Freud’s Adjuvant with 4mg/mL crushed, deactivated mycobacterium tuberculosis and 1mg/mL myelin peptide [MOG 35–55 (C57Bl/6) or PLP139-151 (SJL/J)] were emulsified and injected subcutaneously above shoulders and hips of anesthetized mice, 50 μL per injection site for a total of 200 μL emulsion. For studies using C57Bl/6J mice, pertussis toxin (List Labs, 50μg/mL) in 200μL PBS was diluted to 200–400 ng depending on lot potency and injected intraperitoneal (i.p.) on the day of immunization (day 0) and 2 days postimmunization (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)

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

Shiga toxin work

Shiga toxin 2 (Stx2) was obtained commercially from the USA through List Biological Laboratories Inc. (product #162 lot 1621A1). Stx2 was chosen as it is known to be more cytotoxic in mice than Stx1; and because epidemiological data from human studies have demonstrated that Stx2 producing STEC have a stronger association with more severe disease.22 All work involving the toxin was performed in a biological cabinet under category 3 facilities. Prior to any toxin work commencing, a ‘control of substances hazardous to health regulations’ (COSHH) risk assessment was completed which was submitted and reviewed by the biological and genetic safety committee.

Upon receipt of Shiga toxin, the lyophilized powder was reconstituted with 100μL distilled water and gently mixed by inversion in-keeping with the manufacturer’s instructions. This resulted in a stock concentration of 10μg/100μls which was aliquoted into 20μL stock eppendorfs and stored at −20°C until use. The toxin was assessed for purity on 12% SDS-PAGE and was found to have two major bands at molecular weights of 33 kDa and 8 kDa corresponding to A and B toxin subunits, respectively. Purity was >98% and the endotoxin content determined on kinetic chromogenic LAL assay at List Labs was <90EU/mg. A dose of 10 ng/g of Shiga toxin diluted in sterile normal saline 0.9% (Baxter) was given to each mouse via intraperitoneal injection. All in vitro Shiga toxin cell treatments were performed on differentiated cells at a dose of 0.1 ng/ml.