Bacterial Toxin Research Citations

Intracortical tetanus and botulinum toxin injections:

For intracortical injections, a craniotomy is made over the cingulate cortex of anesthetized mice by thinning the skull with a dental drill and using a fine needle to remove a small (100 m diameter) piece of skull. Using a fine glass capillary mounted to a stereotaxic apparatus, tetanus toxin³² (25ng; 190A, List Biological Labs) or botulinum toxin (0.03ng; 128A, List Biological Labs) was diluted in 7 l of artificial cerebral spinal fluid (ACSF) and injected 500 m into the cingulate cortex.

• Product #190 – Tetanus Toxin from Clostridium tetani
• Product #128 – Botulinum Neurotoxin Type A Complex from Clostridium botulinum

… SNAPtide FITC/Dabcyl substrate, List Biological Laboratories, 521, …

• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin

Experimental autoimmune encephalomyelitis (EAE):

Female C57Bl6/J mice were immunized s.c. at four sites on the flank … On the day of immunization and two days later, pertussis toxin (400 ng, List Biological Labs, CA) was administered i.p. Mice were observed for clinical signs of EAE (paralysis progressing from tail to head) for up to 30 days …

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)

Induction and scoring of EAE:

… Briefly, female mice (68 wk old) were induced for EAE by s.c. injection of a 200 µL IFA/PBS (v/v) solution containing 300 µg MOG peptide and 200 µg of Mycobacterium tuberculosis H37Ra (Difco) in the footpads and at the base of the limbs. Six hours later, mice were given i.v. 500 ng purified Bordetella pertussis toxin (List Biological Laboratories). A second injection of B. pertussis toxin was given after 48 h. The mice were scored daily for clinical signs of EAE as follows: …

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:

… Pertussis toxin (200 ng, List Biological Laboratories, Campbell, CA) was administrated on the day of immunization and 48 h later. For treatment of EAE, compounds 8h or vehicle PBS was orally dosed twice daily from day 0 after immunization onward. …

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:

… Pertussis toxin (200 ng) (List Biological Laboratories) is also injected intravenously on day 0 and 2. Animals are observed daily for clinical signs of EAE for up to 35 days after immunization, and neurological effects were quantified on an arbitrary clinical scale…

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)

Detection of Serum Antibodies to Pertussis Antigens:

Nunc Maxisorp 96-well plates were coated overnight with 0.2 µg/mL PT, 0.5 µg/mL FHA, 2 µg/mL PRN, or 0.2 µg/mL FIM (List Biologicals) as described previously… For whole-bacteria ELISA, plates were coated overnight at 37°C with heat-killed B. pertussis prepared as described above.

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 #186 – Fimbriae 2/3 from B. pertussis
• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

Anthrax toxin neutralization assay (TNA):

The anthrax TNA was performed using the mouse macrophage cell line J774A.1 (European collection of cell cultures, ECACC) as previously described [25] with slight modifications. 96-well flat-bottomed tissue culture plates (Greiner Bio One, Germany) containing 810⁴ macrophages/well in DMEM (Life Technologies, USA) and 10% FCS (Life Technologies, USA) were incubated overnight at 37C and 5% CO₂. Goat sera were doubly diluted (1:50 to 1:6400) in culture medium containing PA and LF (List Biological Laboratories Inc., Campbell, CA) at concentration of 500 ng/ml and 400 ng/ml (lethal toxin, LT) respectively. The sera/LT mixture was incubated for one hour at 37C and 5% CO₂ before adding to overnight cultured cells (after discarding medium) and incubated for three hours. …

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

Adoptive transfer experiments:

Enriched CD4⁺CD25⁺ cells were then cultured in complete medium alone or with pertussis toxin (ptx; List Biological Laboratories) at 100 ng/ml for 2 h in vitro and transferred to congenically marked recipient mice. Splenocytes of recipient mice were harvested 36 h after transfer and analyzed by flow cytometry.

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:

B. anthracis PA protein was from List Biological Laboratories, Inc. (Campbell, CA).

Adsorption of protein antigens on aluminum hydroxide particles:

The adsorption of proteins (OVA or PA) on aluminum hydroxide particles was carried out by mixing the particles in suspension with the protein in solution. Briefly, a certain volume of the protein solution was added into a tube (10 g OVA or 4 g PA), followed by the addition of particles in suspension at a weight ratio of 1:5 to 1:1 (OVA vs. particles) or 1:5 (PA vs. particles). After 20 min of gentle stirring, the protein-particle mixtures were stored at 4C or freeze-dried, if needed, before further use.

Immunization studies:

…When PA was used as the antigen, female BALB/c mice (1820 g) were immunized subcutaneously with PA-adsorbed aluminum hydroxide particles on days 0 and 14. As negative controls, mice were injected with sterile PBS or PA alone. The dose of PA was 4 g per mouse per injection, and the dose of the particles was 20 g per mouse per injection.

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