Bacterial Toxin Research Citations

Induction and assessment of EAE:

For active EAE, mice were immunized subcutaneously on the back with 200 µg of MOG35-55 (MEVGWYRSPFSRVVHLYRNGK) emulsified in CFA (Difco Lab, Detroit, MI) containing 4 mg/ml Mycobacterium tuberculosis H37Ra (Difco). Two hundred nanogram of pertussis toxin (List Biological Lab, Epsom, England) was given intraperitoneally on days 0 and 2 (p.i.). …

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)

… Pertussis toxin (List Biological Laboratories) at a dose of 200 µg per mouse in PBS was administered intravenously on the day of immunization and once more 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)

… Fluorescein isothiocyanate-conjugated B subunit of cholera toxin (CTB-FITC) was purchased from List Biologicals (Campbell, CA, USA). …

Confocal laser microscopic examination of the stable transfectants:

To examine the distribution of GM1 ganglioside (GM1) in these transfectants, we used CTB-FITC as specific probe for GM1. The transfectants were cultured on collagen/poly-l-lysine (25:1)-coated coverslips and fixed with 3% paraformaldehyde for 5 min at room temperature and stained with CTB-FITC.

• Product #106 – Cholera Toxin B Subunit (CTB) from Vibrio cholerae FITC, Conjugate

Tracer injections:

The rats were deeply anesthetized by intraperitoneal injections of a mixture of ketamine (72 mg/kg) and xylazine (11.2 mg/kg). … The retrograde tracers, cholera toxin subunit (CT; List Biological Laboratories, Campbell, CA; 1% in ddH₂O) … The tracer solution was injected ionotophoretically using positive current pulses (7 s on and 7 s off for 15 min) of 1 A (for FG) and 3 A (for CT). In order to generate more retrograde labeling in the vicinity of the RMTg, some injections of CT into RtGi were done with glass pipettes pulled to tip diameters of 3040 m from which the tracer was expelled by air pressure, …

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

Drug Delivery:

All drugs were made in sterile double distilled water (ddH₂0) containing 1% DMSO (100x) unless otherwise noted. Vehicle conditions are cells that were supplied with ddH₂0 and DMSO to a final amount of 0.01% per 3ml of media. After 24 h of transfection, media was removed from cells and replaced with the indicated drug for the length of 48 h unless otherwise noted. In the case of time-course experiments, agonists, modulators and/or secondary drugs (botulinum toxin b) were added for varying durations such that all time-based groups were harvested at the end of the 48 h drug incubation period. This procedure ensured that all groups had equivalent time for protein synthesis and that tissue for immumocytochemistry could be processed in parallel. Final concentrations were as follows: GABA (1 and 10 M), gaboxadol (10 M), -alanine (10 M, 50 M and 1 mM), THP (100 nM) and botulimum toxin B (5 nM). All drugs except for THP (Steraloids, Newport, RI) and botulinum toxin b (List Biological Laboratories, Inc., Campbell, CA) …

… Pressure injections of volumes ranging from 20 to 200 nl of CT (1% low salt solution, List Biologicals, Campbell, CA) were made with a syringe (external diameter 0.24 mm, Hamilton Co., Reno, NV) at a rate of 10 nanoliters per minute (Trojanowski, 1983, Ericson and Blomqvist, 1988). …

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

Materials and methods – Chemicals:

… Cholera toxin was purchased from List Biological Laboratories, Inc. (Campbell, CA, USA). …

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

Experimental autoimmune encephalitis (EAE) induction and clinical scoring:

Six to eight-week-old female mice were immunized subcutaneously with PLP₁₈₀₁₉₉ peptide (200µg/mouse, Genemed Synthesis Inc.) emulsified in CFA containing Mycobacterium tuberculosis H37Ra (500µg/mouse, BD Diagnostics). Mice also received 300ng of pertussis toxin (List Biological Laboratories) intraperitoneally (i.p.) at the time of immunization and 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)

Synthesis of HD5 and Mutant Analogues:

… Recombinant anthrax lethal factor was purchased from List Biological Laboratories, Inc. …

LF Inhibition Kinetics:

The inhibition of LF by various defensins was quantified using an enzymatic kinetic assay (52). Briefly, freshly prepared LF at a final concentration of 1 g/ml (10 nm) was incubated at 37 C for 30 min with a 2-fold dilution series of defensin in 20 mm HEPES buffer containing 1 mm CaCl2 and 0.5% Nonidet P-40, pH 7.2. 20 l of LF substrate (1 mm) was added to each well to a final concentration of 100 m in a total volume of 200 l. The enzyme activity, characterized as a time-dependent absorbance increase at 405 nm due to the release of p-nitroaniline, was monitored at 37 C over a period of 5 min on a 96-well Vmax microplate reader …

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).

Antigen presenting cell cultures:

Adherent splenocytes, and resident peritoneal cells were cultured for 4 or 24 hours in the presence or absence of 100 ng/ml LPS (E. coli O111:B4; List Biological Laboratories, concentration previously determined).  After incubation, supernatants from each culture were collected for analysis by multiplex bead array, and the cells were harvested for analysis by flow cytometry.  

• Product #201 – LPS from Escherichia coli O111:B4