Bacterial Toxin Research Citations

Animal experiment

Animal experimental procedures were approved by the Animal Research Committee of Shandong University Affiliated Shandong Provincial Hospital. The C57BL/6J male mice weighing approximately 25 g were used in this study. … To induce EAE, MOG 35 to 55 peptide (Sigma-Aldrich) was mixed with CFA [incomplete free adjuvant (Sigma-Aldrich) and mycobacterium tuberculosis (BD Difco)], followed by subcutaneous injection of 100ul on each side of the lower back of 6-week-old male C57BL/6J mice. Pertussis toxin (List Laboratories) at a dose of 200 ng was injected intraperitoneally into the mice on days 0 and 2 after immunization. Mice were weighed daily and assessed for clinical symptoms. …

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)

Microglia depletion

For microglia depletion, 80 ng of diphtheria toxin (DT) (0.5 ng/μL, 150, List Labs, USA) was injected intraperitoneally on day -2 and 0 before SAH induction.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

EAE induction

EAE was induced by immunization 8 to 10-week-old female C57BL/6J mice with 250 mg MOG33-55 peptide (Genemed Synthesis)
emulsified in complete Freund’s adjuvant (CFA) (BD Difco) per mouse, injected subcutaneously into each flank, followed by intraperitoneal injection of 150 ng of pertussis toxin (PT) (List Biological Laboratories, Inc.) per mouse on day 0. A second intraperitoneal injection of 300 ng of PT was administered on day 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)

… 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 and scoring

EAE induction was performed by mixing 150 µg rhMOG (Protein Expression Laboratory, Leidos Biomedical Research, Frederick National Laboratory for Cancer Research, Frederick, MD) or 200 μg of MOG35–55 peptide (MEVGWYRSPFSRVVHLYRNGK) (GenScript) in CFA containing Mycobacterium tuberculosis H37Ra (Difco Laboratories). Pertussis toxin (120 ng) (List Biological Laboratories) diluted in PBS was administered by i.p. injection 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)

Diphtheria toxin administration:

For depletion of CD206-expressing macrophages, 500ng (20ng/g body weight, assuming an average 25g weight for each mouse) diphtheria toxin (DTx;List Biological Laboratories) in 100µL 1X PBS was injected intraperitoneally into each mouse – for both Csf1rCre; Mrc1(CD206) LSL-Venus-DTR (DTR) and Mrc1(CD206) LSL-Venus-DTR (WT) groups – at every time point. For the early depletion regime, injections were started 2 days after adoptive transfer of T cells and continued every 2-3 days till endpoint, while for the late depletion regime, injections began at d10 after T cell injection and continued till endpoint. For testing the effects of DTx in tumor-free tissue, similar dosing of DTx as the early depletion regime was implemented without tumor injection, and the skin (ectopic tumor site) and skin-draining lymph nodes were isolated for analysis. Mice were found to be healthy and without frank health issues with 6 doses of 500ng DTx (early depletion regime), but were monitored nevertheless throughout the experiment, as per IACUC guidelines.

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

… In all EAE experiments, mice received 200 ng of pertussis toxin (List Biological Labs) ip at days 0 and 2. Mice were assessed for clinical symptoms daily as previously …

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)

Immunization and in vivo treatment

All immunization procedures were conducted in the rear footpad with 20 µL injection volume per
footpad. Mice were immunized as indicated in each Figure with 5 µg low-endotoxin OVA (Worthington
Biochemical Corporation) mixed with either 50 µg papain (P4762, Sigma), 10 µL FCA (F5881, Sigma),
or 10 µL alum (77161, Thermo Scientific) in phosphate-buffered saline (PBS). For DC depletion, mice
were injected i.p. with 500 ng DT in PBS (List Biological Laboratories)
at indicated time-points. …

T cell priming assays

… Mice with DTR expression were treated with DT as indicated in each Figure. …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

… Pertussis toxin (100 ng per mouse; List Biological Laboratories) was administered intravenously 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)

Bioreagents

Shiga toxin-1 from E. coli (#161, List Laboratories, Campbell, CA, USA), …

• Product #161 – Shiga Toxin 1 from Escherichia coli