Bacterial Toxin Research Citations

Stereotaxic surgery

… For retrograde tracing of TRN-projecting and PVT-projecting PL cells CTB-488 and CTB-555 (1.0% in PBS, Thermo Fisher Scientific) were injected into the TRN (0.5 µL) and PVT (0.6 µL), respectively, and allowed 4 d for retrograde transport. For retrograde tracing of PVT-projecting TRN cells unconjugated CTB (List Labs Product No. 104) was injected into the PVT (0.6 µL). For retrograde labeling of PVT-projecting TRN cells for RNAscope, retrobeads (0.6 µL, LumaFluor, Inc.) were injected into the PVT and allowed 7 d for retrograde transport. …

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

EAE induction

The female C57BL/6 mice (12 weeks old) used for the induction of EAE were purchased from KOATECH (Pyeongtaek, Gyeonggi, Republic of Korea). All mice were housed and maintained in a pathogen-free facility at Seoul National University (SNU) College of Medicine. …To induce EAE, mice received bilateral subcutaneous injections of a total of 200 μg of MOG 35–55 peptide (GeneScript, Piscataway, NJ) emulsified in CFA (BD Biosciences, San Jose, CA), followed by i.p. injection of 250 ng pertussis toxin (PTX) (List Biological Labs, Campbell, CA) 4 and 24 h later. The clinical scores of EAE mice were assessed daily.

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)

Antigens

Overlapping peptide pools targeting ESAT6 and CFP10 were used to stimulate T cells for these studies (BEI Resources). Peptides were 15 or 16 mers with 11 or 12 amino acid overlaps for ESAT6 protein and 11 amino acid overlaps for CFP10. Mtb whole-cell lysate from H37Rv was also used to stimulate T cells (BEI Resources). Dimethyl sulfoxide (DMSO) (Sigma-Aldrich) was used as a negative control. Staphylococcus enterotoxin B (List Biological Laboratories) was used as a positive control for the low-exposure cohort.

… Pertussis toxin (100ng per mouse; List Biological Laboratories) was injected intravenously on days 0 and 2 after immunization. The course of EAE disease was monitored daily, and each mouse was assigned a clinical score according to 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)

Stimulations and viral infections

N/TERT1 cells were plated at 5 × 104 cells/96-well plate for cytokine release experiments and at 2.5 × 105, 5 × 105, or at 1 × 106 cells per 24-, 12-, or 6-well plate, respectively. The next day, the cells were treated as indicated with 2 µM Val-boroPro (ApexBio and Biozol), 2 µM ANS (Biomol), 100 ng/mL TNFα (PeproTech), 6.5 µM Nigericin (Sigma Aldrich), 0.1 µM B/B homodimerizer (AP20187, MedChem Express), 20 µM Z-VAD-FMK (Peptanova), 10 µM cytarabine (Ara C, Sigma Aldrich), 1 µM MLN4924 (MLN), 1 µg/mL anthrax lethal factor, and 1 µg/mL protective antigen (both List Biological Labs). …

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

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

…cholera toxin (CT; 1µg; List Biological Labs; 100B) as a control adjuvant. …

S100A4 as an adjuvant potentiates antigen-specific antibody production after intranasal immunization with the Mtb-derived antigen ESAT-6

Although cell-mediated immune responses are believed to be critical in controlling infection by the intracellular pathogen Mtb, there is evidence that humoral immunity also contributes to protection against TB [26,27]. To confirm the adjuvant activity of S100A4 in enhancing antibody production, we immunized mice with ESAT-6, a candidate vaccine antigen derived from Mtb, alone or admixed to S100A4 or CT, three times at an interval of 10 days. …

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

REAGENT or RESOURCE SOURCE IDENTIFIER
Pertussis toxin List Biological Laboratories, Campbell, CA Cat# 181238A1

Preparation of pertussis toxin (PTx) solution
Timing: 5 min

3.
Dilute one vial of PTx toxin derived from Bordetella pertussis lyophilized powder (50 μg) in 50 mL of phosphate-buffered saline (PBS) to obtain a 1 μg/mL PTx solution. After dissolution, this solution can be stored in a refrigerator at 4°C for at least 6 months.

Intraperitoneal injection of PTx toxin
Timing: 2–3 min per mouse

7.
After 2 h of MOG injection, administer 200 μL of pertussis toxin (200 ng per mouse) on the same day via the i.p. route.
8.
Repeat the pertussis toxin administration following 48 h of MOG35-55 immunization.

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

Induction and treatment of EAE

The EAE animal model was developed as previously described [19]. In brief, male C57BL/6 mice were immunized subcutaneously with 200 µg MOG35 − 55 peptide (MEVGWYRSPFSRVVHLYRNGK, GL Biochem, Shanghai, China) emulsified in Complete Freund’s adjuvant, which contains 5 mg/ml heat-killed Mycobacterium tuberculosis (H37Ra strain, BD Diagnostics). Each mouse received a total volume of 200 µl of MOG35 − 55 peptide/CFA emulsion, with 50 µl injected subcutaneously at two separate sites on the back, resulting in four injections per mouse. Pertussis toxin (dissolved in sterile distilled water, 200 ng/mouse, List Biological Labs) was injected intraperitoneally on day 0 and day 2 following 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)