Citations

Enzyme-linked immunosorbent assay

Nunc-immuno microtitre plate wells with Maxisorp surface (Thermo Fisher) were coated in triplicates with 30ng/well of full-length TcdA (List Biological Laboratories, Inc., Campbell, CA) in 0.05M carbonate–bicarbonate buffer pH 9.6, and incubated overnight at 4°C. Wells were blocked with 1% (w/v) bovine serum albumin in PBS for 1h at room temperature, washed with PBS/0.05% (v/v) Tween 20 and then incubated for 1h with decreasing concentrations of His6-tagged VHH constructs in PBS containing 0.2% bovine serum albumin and 0.05% Tween 20 (PBS-TB). …

• Product #152C – Toxin A from Clostridium difficile

… Unless otherwise specified, mice were sensitized by a single intraperitoneal injection of 250 µg ovalbumin (Sigma; A5478) and 5 µg cholera toxin (List Labs; 100B) in PBS …

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

… For oral sensitization experiments, mice were given oral gavages on days 0 and 7 with 5 mg of grade III OVA with 0.5 mg/kg of cholera toxin (List Biologicals 100B, lot nos. 10165A1 and 10165A2) in a final volume of 0.25 mL 0.2 M sodium bicarbonate buffer. …

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

… All mice were anesthetized with isoflurane (2%) in 100% oxygen as described previously, while injections of CTb 1% (low salt, 1%, List Biological Laboratories, Campbell, CA) in distilled water, were made using a glass micropipette (tip diameter 15-20 µm) and Nanoinject II, to retrogradely …

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

… CTb was detected using a polyclonal goat anti-CTb (List Laboratories; …

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

… Pertussis toxin (200 ng, List Biological Laboratories) was injected intraperitoneally on days 0 and 2. …

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)

… 100 ng/ml cholera toxin (List Biological Labs), …

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

2.1 Animals and EAE induction

C57BL/6 female mice, 8-10 weeks of age (18-20 g), were obtained from SPF Biotechnology Co., Ltd. (Beijing, China), and housed in specific pathogen-free conditions at the Fifth Medical Center of PLA General Hospital (animal ethics committee approval No. YFYDW2020017), Beijing, China. All efforts were made to reduce animal suffering, and the Institutional Committee on Care and Use of Research animals approved the procedures used in this study. EAE was induced as described previously (Dou et al., 2021). Inactivated Mycobacterium tuberculosis (#231141, Difco) was added with Freund’s incomplete adjuvant (#F5506, Sigma) to prepare a 10 mg/mL solution, which was fully mixed with an equal volume of myelin oligodendrocyte glycoprotein peptide (MOG35-55, MEVGWYRSPFS RVVHLYRNGK; GenScript) solution (2 mg/mL) dissolved in sterile phosphate-buffered saline (PBS) to prepare water in oil antigen emulsion. Mice anesthetized with pentobarbital at four points on both sides of the midline of the head and back, a total of 0.2 mL/mouse. On 0 h and 48 h post-immunization (p.i.), 200 ng/mouse pertussis toxin (#180, List Biological) was injected intraperitoneally.

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

… adjuvant including heat-killed Mycobacteria tuberculosis (BD Difco, #DF3114-33-8, 200 lg/mouse) on day 0, along with ip injection of pertussis toxin (List Biological 181, …

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

… pertussis Lyophilized (Salt-Free)List Labs Catalog ##181 …

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