Bacterial Toxin Research Citations

Materials and Methods

A total of 40 female C57BL/6 mice, 10-week-old, were randomly divided into four groups, including induced control groups without EAE induction, induced EAE without training, and forced and voluntary training groups. All animals have started training 4 weeks before experimental autoimmune encephalitis- (EAE-) induction. The mice were intraperitoneally anesthetized with an injection of xylazine (5 mg/kg) and ketamine (50 mg/kg). Myelin oligodendrocyte glycoprotein peptide 35–55 (300 μg in saline; MOG 35–55; KJ Ross-Petersen ApS, Denmark) was emulsified in an equal volume of complete Freund’s adjuvant (CFA; Sigma, USA) containing 500 μg of heatkilled Mycobacterium tuberculosis. This emulsion was then injected subcutaneously at the base of the tail of each mouse. The mice also received intraperitoneal injection of pertussis toxin (300 ng in 10 μl PBS; List Biological Lab, USA) at the time of immunization and the same was repeated 48 hours later. To confirm EAE induction, animal clinical assessment of EAE was performed daily using a 15-point scoring system following immunization as reported previously by Giuliani et al. [13].

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)

Animal care and EAE immunization

… Mice received intraperitoneal (I.P.) injections with 400 ng (C57BL/6) or 200 ng (FoxP3EGFP) of Bordetella pertussis toxin (List Biological Laboratories) on day −21 and day −19 prior to transplant. …

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)

… and after rats resumed normal growth (required as evidence of recovery from surgery), they are briefly anesthetized to inject 2.5 ng TeNT (1 L in phosphatebuffered saline with 2% bovine serum albumin; produced by List Biological Inc …

• Product #190 – Tetanus Toxin from Clostridium tetani

… Prior to experimental manipulation, mice were anesthetized by intraperitoneal injection of ketamine (100 mg/kg) and xylazine hydrochloride (10 mg/kg). For intranasal or ocular immunization, 1 μg of TLA or TLA exo Ags with 2 μg cholera toxin (CT) (List Biological Laboratories, Campbell, California, USA) per head was suspended in 10 μl PBS and dropped 3 times at 2 week interval onto the surface of the cornea making a droplet or intranasally onto the nostrils using a micropipette. …

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

… All dosing solutions were prepared in DMEM from stocks immediately prior to dosing for final dosing concentration of 40M or 10M and 0.0004% DMSO. Stocks were prepared of ultrapure lipopolysaccharide (LPS; 1 endotoxin unit (EU)/ng, E. coli 055:B5 Lot#4231A1 List Biological Laboratories, Campbell, CA) with sonication in nuclease free sterile water …

• Product #423 – ULTRA PURE LPS from Escherichia coli O55:B5

… On day 0, the mice were also administered 500 ng of pertussis toxin (List Biologicals, Campbell, CA, USA) intraperitoneally. …

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)

… a group of Wistar rats received unilateral microinjections of the retrograde tracer Cholera Toxin B subunit (CTB; 1% w/v in artificial cerebrospinal fluid (ACSF), 100 nL, List Biological Laboratories, Campbell, CA, USA) into the BLA (all coordinates are expressed in mm from bregma, AP: 1.9; ML: 5.00; DV: 8.5) in experiment 1 …

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

Bioassay Reagents

… Active CT produced in V. cholerae was purchased from List Biological Laboratories, azide-free in powder form (product no. 100B). …

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

EAE immunization and scoring

As described previously27,28,29,30, mice were immunized s.c. on d0 in the right and left flank with 100ug of PLP antigen emulsified 1:1 volume in complete Freund’s adjuvant supplemented with 4 mg/ml Mycobacterium tuberculosis (CFA; Becton Dickinson, Franklin Lakes, NJ), followed by 250 ng pertussis toxin (PTx; List Biologicals, Campbell, CA) i.p. 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)

EAE induction, treatment and assessment. Mice were immunized at multiple sites by subcutaneous injection of 200 μg of MOG35-55 or 200 μg of OVA protein (as a control) (Sigma-Aldrich) emulsified in 200 μl of complete Freund’s Adjuvant supplemented with 5 mg/mL heat-inactivated Mycobacterium tuberculosis H37Ra (Thermo Fisher). A total of 200 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) was injected intravenously into the mice after the MOG3555 injections and again 2 days 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)