Bacterial Toxin Research Citations

EAE induction:

EAE was induced in WTand Kv1.3 KO mice by s.c. injection over two sites in the ank with 100 mg myelin oligodendrocyte glycoprotein (MOG 35 55) peptide (The Johns Hopkins Peptide Synthesis Core Facility) emulsied in CFA (Difco, Detroit, MI). A total of 250 ng pertussis toxin (List Biologicals, Campbell, CA) was administered at the time of immunization and 48 h later by i.p. injection. Mice were monitored daily for clinical signs of EAE and given a clinical score based on the following criteria: …

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 induction by active immunization:

For EAE induction, female WTand GILT2/2 mice were immunized by s.c. injections with either 300 mg MOG3555, or 100 mg rMOG proteins in CFA (Difco, Detroit, MI) with 300 mg Mycobacterium tuberculosis on days 0 and 7. Mice received i.p. injections of 500 ng pertussis toxin (PTX) (List Biological Laboratories, Campbell, CA) 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)

Animals and surgical procedures:

Adult male Harlan SpragueDawley rats … were anesthetized with an equal mixture (1 ml/kg body weight) of ketamine (50 mg/ml) and xylazine (10 mg/ml), delivered intramuscularly; supplements of 0.5 ml ketamine were given if required…

Under anesthesia, each animal received a single, stereo-taxically placed iontophoretic co-injection of a mixture of 2.5% PHA-L (#L-1110; Vector, Burlingame, CA) and 0.25% CTB (#104; List Biological, Campbell, CA) prepared in 0.1 M sodium phosphate-buffered saline (pH 7.4). …

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

Recombinant protein reagents:

Recombinant anthrax toxins, rPA, rLF, or rEF (List Biological Laboratories, Campbell, CA) were stored frozen in 1 mg/ml aliquots as recommended by the supplier. …

Protective antigen, lethal factor, and edema factor ELISAs:

Antibody specificity for toxin components was determined using standard ELISAs. High bind plates (Costar 3369) were coated overnight with 1 g/well of rPA, rLF, or rEF. …

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).

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

Author did not specify which List Labs Anthrax Edema Factor was utilized.  List Labs provides Product #178A(Anthrax Edema Factor (EF), Recombinant from B. anthracis) and Product #173B (Anthrax Edema Factor (EF), Recombinant, Mutant S447N from B. anthracis).

… To examine the mucosal immunogenicity of MucoRice-BoHc, … with or without CT (1 µg, List Biological Laboratories, Campbell, CA) …

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

Induction of EAE:

EAE was induced by subcutaneous immunization of eight-week-old mice in the flanks with 200µl of an emulsion containing 250µg of MOG₃₅₅₅ peptide (MEVGWYRSPFSRVVHLYRNGK) and 400µg of Mycobacterium tuberculosis extract H37 Ra (BD Biosciences, Austin, TX, USA) in incomplete Freunds adjuvant oil. In addition, the animals received intraperitoneal injection of 250ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA). on Day 0 and 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)

… In some experiments, inhibitors were injected intracellularly into the sensory neuron 30 min before the start of an experiment, and were always compared with vehicle injections. EGTA (Calbiochem) and the light chain of botulinum toxin D (List Biological Laboratories) were diluted in the vehicle solution (0.5 M KAc with 10 mM TrisHCl, pH 7.4, and 0.2% Fast Green to visualize the injection).

Regents:

 … the light chain of botulinum toxin D (List Biological Laboratories), and sense and antisense oligonucleotides were diluted in the vehicle solution (0.5 M KAc with 10 mM TrisHCl, pH 7.4, and 0.2% Fast Green to visualize the injection). …

Surgery:

… To verify that visual behaviors were mediated by the regenerating optic nerve, we reanesthetized mice in Groups I and II at 7 wk and transected the previously intact right optic nerve. Five days before the end of the study, we injected CTB (3 µL, 1%; List Biological) as an anterograde tracer to visualize axons and nerve terminals originating in RGCs. Most mice in the behavioral study survived for 10 wk but a few survived 12 wk. …

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

Synthesis and Folding of Defensins:

… Recombinant LF was purchased from List Biological Laboratories, Inc. …

LF Activity Inhibition by l-Ala17-HNP1 Was Less Than Wild-type HNP1:

The enzyme activity of LF in the presence of HNP1 or l-Ala17-HNP1 was assessed using a method adapted from Kaufmann and colleagues (30). Fig. 7 shows that the inhibitory activity of l-Ala17-HNP1 as measured by the concentration that gave 50% inhibition (IC50) differed slightly more than 2-fold compared with HNP1. By contrast, d-Ala17-HNP2 (HNP1 numbering) was equally active compared with HNP2. The d-Ala17 mutation was incorporated into HNP2 instead of HNP1 to conform to the previous study (18), but because HNP1 and HNP2 vary only by the deletion of a single Ala residue at the N terminus, they should be comparable with one another with respect to the internal Gly17 residue. LF inhibition is impaired only by the Gly17 to l-Ala17 mutation but not the Gly17 to d-Ala17 mutation, pointing to a local structural defect as the culprit.

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).