Citations

EAE

Mice were immunized subcutaneously with 100 μg of myelin oligodendrocyte glycoprotein (MOG)35–55 peptide emulsified in CFA, which was made from IFA (Difco) plus Mycobacterium tuberculosis (Difco). Mice were injected intraperitoneally with 200 ng of pertussis toxin (PTX, List Biological Laboratories) on the day of immunization and 2 d later. …

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)

Reagents

Pertussis toxin (PTX) was obtained from List Biological Labs.

Induction of EAE

MOG35-55-induced EAE in C57BL/6 mice was established as previously described (11–13). In brief, the mice were immunized on day 0 with an intracutaneous injection of 150 mg of MOG35-55 emulsified in CFA-bearing M. tuberculosis strain H37Rv. Injection volume was set at 150 µl. Thereafter, each mouse received an additional 300 ng of PTX by intraperitoneal injection in 100 ml of phosphate-buffered saline (PBS) on day 0 and again on day 2 post-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)

Food allergy model

… For the peanut-induced systemic anaphylaxis model50, C3H/HeJ mice were intragastrically sensitized with 10 mg peanut extract (GREER Laboratories, USA) and 20 μg cholera toxin (List Biological Laboratories, USA) 4 times weekly. The animals were then orally boosted with 50 mg peanut extract and 20 μg cholera toxin twice every 2 weeks. At week 8,200 mg peanut extract was orally administered and rectal temperature was measured. In this model, IgETRAP was intraperitoneally injected the day before challenge of 200 mg peanut, and 5 × 109 cfu of B. longum was intragastrically administered to each mouse (5 × 109 cfu head−1) every day from week 5 to 8. …

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

… Cholera Toxin List Biological laboratory 100B …

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

Induction and assessment of EAE

Acute EAE was induced and assessed as previously described (Kang et al, 2010). Briefly, acute EAE was induced by subcutaneous immunization with 300 μg of the MOG35-55 peptide (Met-Glu-Val-Gly-Trp-Tyr-Arg-Ser-Pro-Phe-Ser-Arg-Val-Val-His-Leu-Tyr-Arg-Asn-Gly-Lys, Sangon Biotech Co.) in CFA containing 5 mg/ml heat-killed H37Ra strain of Mycobacterium tuberculosis (Chondrex, Inc) in the back region and both sides of the vertebrae. And the immunized mice were i.v. injected with pertussis toxin (List Biological Laboratories, Inc.) at a dose of 250 ng per mouse in PBS on the day of immunization and once more 48 h after the first injection. …

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)

2.4. Immunohistochemistry

Following transcardial perfusion, the brains were dissected and postfixed in 4% PFA for 1.5 h at room temperature (RT). Subsequently, the brains were transferred to 10% sucrose in 0.1 M PB and left overnight at 4 °C, before being embedded in gelatin. The brains were incubated in 12% gelatin/10% sucrose in 0.1 M PB for 30 min at 37 °C, embedded in a plastic mold and left at 4 °C for at least 30 min to harden. They were then cut in small blocks and placed in a 10% formalin/30% sucrose in 0.1 M PB for at least 2.5 h at RT. Embedded brains were transferred to 30% sucrose in 0.1 M PB overnight at 4 °C before being sliced on a Leica SM2000R microtome (Leica Biosystems, Nussloch, Germany) at 50 µm. To prepare the slices for antibody staining, they were blocked for one hour with 10% normal horse serum (NHS) and 0.5% triton in PBS at RT after rinsing in PBS. Subsequently, slices were stained for goat anti-CTB (703; 1:15,000, List Biological Laboratories, Inc., Campbell, NJ, USA) in 2% NHS and 0.4% triton in PBS, and left 24 h at RT. The CTB antibody turned out to be specific enough for unambiguous identification of intensely labeled CTB+ cells on 10× versus background (intrinsic) fluorescence (for which controls have been previously described by our lab [31]). …

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

Induction of EAE

The EAE model was performed in the “Salari Institute of Cognitive and Behavioural Disorders (SICBD)”. The animal model was induced by injection of MOG35-55 (MEVGWYRSPFSRVVHLYRNGK) as is previously described51. Briefly, Under the anaesthesia caused by ketamine hydrochloride (50 mg kg − 1; Alfasan, Woerden-Holland) plus Xylazine (5 mg kg−1; Alfasan, Woerden-Holland), mice were immunized with 300 µg of myelin oligodendrocytes glycoprotein (MOG35-55; Sigma-Aldrich, United States) dissolved in phosphate-buffered saline (PBS) and emulsified with an equal volume of complete Freund adjuvant (CFA; 400 µg of Mycobacterium tuberculosis; Sigma Co, USA). Additionally, 300 ng of pertussis toxin (List Biological Labs, Campbell, CA, USA) was injected intraperitoneally, into all animals, 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)

… LPS from E. coli O111:B4 (List Biological Labs #201) …

• Product #201 – LPS from Escherichia coli O111:B4

Detection of Shiga toxin 2

Detection of Stx2 in the E. coli O157:H7 culture supernatants and in fecal samples from mice, described below, was carried out by ELISA, as previously described.38 Briefly, camelid anti-Stx antibody (List Biological Laboratories, Campbell, CA) was coated on a white MaxiSorp nunc plate (Thermo Fisher Scientific, Rockford, IL) and incubated overnight at 4°C. …

… Goat anti-PA (B. Anthracis) List Biological Laboratories Cat# 771B …