Bacterial Toxin Research Citations

Evaluation of i.v. administration of BLG-p(Man) in a mouse model of BLG sensitization

This BLG sensitization model was adapted from ref. 27 and 28. Beginning at 2 weeks of age, SPF C3H/HeJ mice were treated with a
mixture of antibiotics (Abx). For the first week before weaning, mice were treated with 100 mL Abx solution once daily, including kanamycin (4 mg/mL), gentamicin (0.35 mg/mL), colistin (8500 U/mL), metronidazole (2.15 mg/mL), and vancomycin (0.45 mg/mL)
(Sigma-Aldrich). After weaning, mice were given drinking water supplemented with a 50-folded dilution of the same Abx solution
except vancomycin, which was maintained at 0.45 mg/mL, throughout the duration of the experiment. Prior to weaning, on day 1
and day 5 after first oral gavage of Abx, the mice were i.v. administered with PBS, or 2 mg BLG in the form of unmodified (free)
BLG or BLG-p(Man) per 10 g body weight. Right after weaning, mice were sensitized weekly for 5 weeks by intragastric gavage
with BLG (20 mg per 10 g body weight) (Sigma-Aldrich) and cholera toxin (CT, 10 mg per 10 g body weight) (List Biologicals, Campbell, CA). …

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

Recombinant antigens

Pertussis toxin (salt free) was obtained from List Labs (Campbell, CA, USA), …

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

SF stimulation

SF were stimulated with human recombinant TNF (10 ng/ml; R&D Systems), human recombinant IL1β (1 ng/ml; R&D Systems), lipopolysaccaride (LPS) from Escherichia coli J5 (100 ng/ml; List Biological Laboratories), polyI-C (PIC) (10 μg/ml; InvivoGen), bacterial lipopeptide (bLP) palmitoyl-3-cysteine-serine-lysine-4 (300 ng/ml; InvivoGen) or human recombinant TGFβ (10 ng/ml; R&D Systems) for 24 h. …

• Product #301 – LPS from Escherichia coli J5 (Rc)

EAE induction

For EAE induction, mice were injected subcutaneously at two sites on the back with 200 mg MOG35–55peptide (synthesised by Toray Research Center, Tokyo, Japan) and 1 mg heat-killed Mycobacterium tuberculosis H37RA emulsified in complete Freund’s adjuvant (Difco, KS, USA). Four hundred nanograms of Pertussis toxin (List Biological Laboratories, CA, USA) were injected intraperitoneally on days 0 and 2 after immunisation. …

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)

Experimental autoimmune encephalitis

EAE was induced as described previously (33). Female C57BL/6 mice (9 weeks of age) were immunized by subcutaneous injection with recombinant MOG35–55 (200 μg, Biomatik Corporation, Cambridge, ON, Canada) emulsified in complete Freund’s adjuvant (incomplete Freund’s adjuvant, Sigma-Aldrich, Oakville, ON, Canada; 4 mg/ml Mycobacterium tuberculosis, strain H37Ra, BD Biosciences, Missisauga, ON, Canada) into four sites on the back (50 μl per site). Pertussis toxin (200 ng, List Biological Laboratories Inc., Campbell, CA, USA) was injected intraperitoneally on days 0 and 2 after 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)

2.2. Induction of EAE

rMOG (2mg/ml) was emulsified with an equal volume of complete Freund’s adjuvant (CFA, 4mg/mL, Sigma, F5506). EAE animal model was induced by subcutaneous injection of the above emulsion into the back. 300ng pertussis toxin (List Biological Laboratories, 180228A1) was injected into the tail vein on the day of immunization and the second day after immunization. Female mice under the same conditions were used as control, and CFA was injected subcutaneously. …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… For CT-B processing, sections were blocked in 10% horse serum (2h, room temperature) and incubated in anti-CT-B made in goat (List Biological #703; 1:4000 dilution) in TBS-TX/3% horse 234 serum for 2-3 days at 4oC. After thorough washing, sections were further incubated in ImmPRES …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Anti-Cholera Toxin B Subunit (Goat) List Biological Laboratories Cat#703; …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

4.4. EAE Induction and Clinical Score Assessment

Active EAE was induced as previously described [34]. Briefly, mice were subcutaneously injected with 200 μg of MOG35–55 peptide (Genscript, RP10245) emulsified in incomplete Freud’s adjuvant (Sigma, F5506; St. Louis, MO, USA) with mycobacterium tuberculosis H37Ra (BD Difco, BD 231141; Pittsburgh, PA, USA) and intraperitoneally injected with 200 ng of Pertussis Toxin (List Biological Laboratories, Campbell, CA, USA). …

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)