Bacterial Toxin Research Citations

2.3 Experimental autoimmune encephalomyelitis

Mice were induced with experimental autoimmune encephalomyelitis (EAE) following a protocol well established in the laboratory (Brambilla et al., 2011; Madsen et al., 2020). Briefly, on day 0 and 2, pertussis toxin (PTX, List Biological Laboratories, #181) diluted in PBS was injected intraperitoneally (500 μg/mouse). On day 1, myelin oligodendrocyte glycoprotein peptide 35–55 (MOG35–55, Bio-Synthesis, #A9349-1) emulsified in complete Freund’s adjuvant was injected subcutaneously (0.33 mg/mouse). …

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

Recombinant antigens

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

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

… Pertussis toxin (PTX, cat: 181) was purchased from List Biological Laboratories (CA, USA). …

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

… Animals were given two 400 ng intraperitoneal injections of B. pertussis toxin (Cat# 181; List Biological Laboratories) on days 0 and 2 …

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

… For EAE alone, mice were immunized with 100 µg of MOG35–55 (Genemed synthesis, Inc) in an emulsion containing complete Freund’s adjuvant (CFA, Difco), then injected peritoneally with 200 ng of Bordetella pertussis toxin (lot# 181236A1, List Biological Laboratories) on post-immunization day 0 and 2. For CPZ/EAE treatment, Abcd1y/− and wild-type mice were fed 0.2% CPZ diet for 2 weeks followed by MOG immunization18. We performed two separate rounds of CPZ alone and EAE alone treatments, and we also conducted CPZ/EAE combined treatment at three different time points. We used 2–3 mice per group in each round. …

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

EAE induction

Experimental autoimmune encephalomyelitis (EAE) was induced in eight week-old female C57BL/6 mice via immunization with myelin oligodendrocyte glycoprotein (MOG)35–55 (PolyPeptide Laboratories). 200 μg MOG35–55 was dissolved in complete Freund’s adjuvant containing 400 μg of Mycobacterium tuberculosis (231141, Difco) and injected subcutaneously in 0.2mL volume per mouse. Pertussis toxin (181, List Biological labs Inc) was administered via intraperitoneal injection after the MOG 35–55 injection and two days later at 75ng and 200ng per mouse doses, respectively.

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

… Pertussis toxin was acquired from List Biological Laboratories (#181, Campbell, Santa Clara Country, California, USA) …

2.2. EAE Induction and Clinical Scores

Mice (female, strain C57BL/6J, 18–20 g, 6–8 weeks) were immunized following a standard protocol, with minor modifications [24]. Briefly, animals were administered incomplete Freund’s adjuvant containing 4 mg/mL Mycobacterium tuberculosis (strain H37Ra) and 200 μg of the myelin oligodendrocyte protein 35–55 (MOG35–55). This was followed by intraperitoneal administration of 400 ng of Pertussis toxin on the day of immunization and two days after. …

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

EAE introduction and evaluation

Each mouse was injected subcutaneously with a 0.2 mL emulsifier containing 50% MOG35 − 55 (Genescript, USA) and 50% complete Freund’s adjuvant (CFA; Sigma‒Aldrich, USA) at the posterior neck and root of the tail on day 0. The emulsifier was prepared from 200 µg of MOG35 − 55 dissolved in 0.1 mL of PBS with an additional 4 mg/mL Mycobacterium tuberculosis H37Ra (Sigma‒Aldrich, USA). At the same time, 300 ng of PTX (List Biological Laboratories 181, USA) was injected intraperitoneally on days 0 and 2. …

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

… Adjuvant, complete H37 Ra VWR Cat#231131 Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free) List Biological Laboratories, Inc. Cat#181 …

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

… Immunization was carried out at 0, 7, 14, 21 and 28 days. In the EAM-PTX (+) group, PTX (List Labs, cat. #: 181, Campbell, California, USA) 2 µg was intraperitoneally injected on days 0 and 7. …

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