Bacterial Toxin Research Citations

Induction of Experimental Autoimmune Encephalomyelitis (EAE):

…The day of MOG immunization was designated day 0. On day 0 and day 2 post-immunization (dpi), mice were injected intraperitoneally with 500 ng Pertussis toxin (List Biological Laboratory). … 

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)

Materials and methods:

We used 16 male Sprague-Dawley rats weighing 250300 g. … To investigate the distribution of vagal motor neurons in the medulla oblongata projecting to the gastrointestinal tract, Fluorogold (Flurochrome LLC, Denver, CO) was applied to the cut end of the accessory celiac branch or the left gastric branch of the vagal trunk at the ventral wall of the subdiaphragmatic esophagus. CTb (List Biological Laboratories, Campbell, CA) was then injected into the body of stomach wall or the duodenum of the same animal. … Then two tubes were fixed with cyanoacrylate glue. Using a glass micropipette (tip diameter = 100 µm) affixed to a 10 µl Hamilton syringe, the same animal was injected with 10 µl of 1% CTb by pressure into the ventral wall of the body of stomach, including the cardia, or the ventral wall of the duodenum about 1 cm aboral to the pylorus. … we applied tracers to the left vagal branches or the ventral wall of the stomach and the duodenum, and then examined the DMV. For control experiments, we poured 10 µl of 2% Fluorogold or 1% CTb into the abdominal cavity. Three days after the application of tracers, the animals were perfused with saline followed by 500 ml of 4% paraformaldehyde-15% picric acid in 0.1 M phosphate buffer. The medulla oblongata was removed, and serial sections were made at a thickness of 70 m using a Vibratome. The sections were incubated with goat anti-CTb (#703, List Biological Laboratories, Campbell, CA; 1:5,000) for 1 day, …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

EAE induction and treatment:

… For EAE induction, mice were challenged subcutaneously in the flank with 200 g of myelin oligodendrocyte glycoprotein (MOG)₃₅₅₅ peptide … On days 0 and 2, mice were treated by intraperitoneal (i.p.) injection with 200 ng of Bordetella pertussis toxin (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)

hSCID Chimeras and In Vivo Plasmablast Expansion:

Antigens used in this study include Tetanus toxoid (List Biological Laboratories)…The 6- to 8-week-old female SCID/beige mice were purchased…

• Product #191 – Tetanus Toxoid from Clostridium tetani 

… Beginning at 14 days of age and again on days 21 and 35, pigs received an IP injection of 100 µg of purified Ovm (prepared in lab, Rupa et al., 2008a) adjuvanted with 10 µg of cholera toxin (List Biologicals, Campbell, CA). …

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

Induction and evaluation of EAE:

Mice were immunized for the induction of EAE, using either a 13 or a 23 immunization protocol…On the day of immunization each mouse received 200 ng pertussis toxin (PTX) i.v. (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)

2.3. Inoculum and administration

… Native, sterile, preservative-free biologically active cholera toxin (List Biological Laboratories, Inc.) was used. Based upon spread plate cultures, the thawed inoculum contained 2.0 × 1010 CFU/dose and also contained approximately 2× that number of dead organisms. Cholera toxin (50 μg) was added to the bacterial inoculum (Lot ELH072910), and a 1 mL volume pipetted onto the absorbent pad of a Tegaderm patch (3 M Company) (Fig. 1). Preclinical cutaneous toxicology studies were performed using CT and the bacterial strain in animals by both the oral and cutaneous routes.

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

…In groups 1, 2, 3 and 5, vaccine formulations consisted of Hank’s balanced salt solution (HBSS) with 5 µg cholera toxin (CT) (List Biological Laboratories Inc., Madison, NJ, USA), …

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

Materials and Methods:

Recombinant, nicked PA₆₃ as well as LF from B. anthracis were obtained from List Biological Laboratories Inc. (Campbell, CA, U.S.A). One mg of lyophilized PA₆₃ protein was dissolved …

Cell Culture, Cytoxicity Assay and Measurement of TNF-release:

…Cells were trypsinized and reseeded for at most 15-20 times. For cytotoxicity experiments, cells were seeded in 96-well-plates and incubated with PA₆₃+ LF together with the respective heterocyclic azolopyridinium salt in serum-free medium. After the indicated incubation periods, the morphology of J774A.1 cells was visualized by using a Zeiss Axiovert 40CFl microscope (Oberkochen, Germany) with a Jenoptik progress C10 CCD camera (Jena, Germany). The cytotoxic effect caused by PA₆₃/LF on this cell line was analyzed in terms of cell lysis. …

Membrane Translocation Assay:

The pH-dependent membrane translocation of LF through PA₆₃-channels was investigated on the surface of intact J774.A1 cells as originally described earlier (1, 2). Cells were grown in 96-well-plates and incubated for 30 min at 37C in serum-free medium with bafilomycin A1 (100 nM). During this period, some cells were treated also with 100 M of either chloroquine-based heterocyclic azolopyridinium salts (HA substances). Subsequently, the cells were incubated for 30 min at 4C with PA₆₃ (1 g/mL) plus LF (1 g/mL) to enable binding of these proteins to the cell surface. Thereafter, the medium was removed and cells were incubated for 5 min at 37C with acidic medium (pH 5.0, for control pH 7.5) containing the respective HA substance (100 M). The acidic medium was removed and cells were incubated at 37C in neutral medium containing bafilomycin A1 (100 nM) to prevent normal uptake of LF. Pictures were taken after 2 h and the toxin-induced cell lysis was analyzed exactly as described before.

Lipid Bilayer Experiments:

Black lipid bilayer experiments were performed as described previously [63] using a 1% solution of diphytanoyl phosphatidylcholine (Avanti Polar Lipids, Alabaster AL, U.S.A.) in n-decane as membrane forming lipid. The instrumentation consisted of a Teflon chamber with two aqueous compartments separated by a thin wall. The small circular whole between the two compartments had a surface area of about 0.4 mm². The aqueous salt solutions were buffered with 10 mM MES, pH 6. All salts were obtained from Merck (Darmstadt, Germany). PA₆₃ was added from concentrated stock solutions after the membrane had turned black, to the aqueous phase to one side (the cis-side) of the membrane. The PA-induced membrane conductance was measured after application of a …

Titration Experiments

The binding of heterocyclic azolopyridinium salts to PA₆₃-channels was studied by titration experiments similar to those used previously to investigate binding of carbohydrates to the LamB-channel of Escherichia coli or binding of chloroquine or EF and LF, respectively, to C2II- and PA₆₃-channels in single- or multi-channel experiments [27], [31], [33], [42], [64]. PA₆₃-channels were reconstituted into lipid bilayer …

In most cases the different HA-substances caused full blockage the PA₆₃-channels. In case of only partial blockage of the PA₆₃-channels caused by a certain fraction of channels that did not respond to binding …

• Prouct #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

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 #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

Antigens and EAE Induction:

…Each mouse received 0.05 ml emulsion (s.c.) at four sites and pertussis toxin (300 ng i.p.; List Laboratories, Campbell, CA) at 0 h and 72 h 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)