Bacterial Toxin Research Citations

Animals, induction of cholangitis and Baricitinib treatment

Adult male C57BL/6JCrl mice aged 8–10 weeks were purchased from Charles River (Shanghai). Mice were housed under specific pathogen-free conditions at the Zhejiang University. … Murine model of autoimmune cholangitis was established as M. Eric Gershwin reported53, 100 μg 2OA-BSA (in 50 μl PBS) were emulsified with 50 μl of Complete Freund’s Adjuvant (CFA, Sigma-Aldrich) and injected intraperitoneally into 6-week-old female mice. Two days after the initial immunization with 2OA-BSA, mice received 100 ng of pertussis toxin (List Biological Laboratories) in 100 μl PBS by I.P. After two weeks, the mice were re-boosted with 100 μg 2OA-BSA (in 50 μl PBS) emulsified with 50 μl of Incomplete Freund’s Adjuvant (IFA, Sigma-Aldrich) I.P. Mice received, baricitinib (10 mg/kg) in 0.5% methylcellulose, or vehicle only (0.5% methylcellulose), as indicated, once daily per oral gavage 6 weeks after initial 2OA-BSA immunization for 2 weeks. The dosage of baricitinib applied were based on previous works, as exemplified by ref. 54.

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)

… Diptheria toxin List Biological Laboratories Cat#150 …

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

CD4+T and YFP+ Treg cells adoptive transfer in Experimental autoimmune encephalomyelitis (EAE)

CD4+YFP+ Treg cells from Foxp3cre and Foxp3cre Cxxc1fl/fl mice were enriched using the Mouse CD4 T Cell Isolation Kit (480005; Biolegend) and then sorted using the BD Aria II flow cytometer. Naïve CD4+T cells from 2D2 (MOG35-55-specificTCR transgenic) mice were isolated using the Mouse CD4 Naïve T cell Isolation Kit480039 (480039; Biolegend). … One day after cell transfer, the recipient mice were inoculated subcutaneously (s.c.) with 200μg MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK; GenemeSynthesis) emulsified in complete Freund’s adjuvant (CFA) (F5506; Sigma). Intravenous administration of 200 ng of Pertussis toxin (181; List Biological Laboratories) was performed on days 0 and 2 after peptide inoculation. …

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

… and 0.25% CTB (List Biological Labs, Inc., Campbell, California, USA; catalog #104) dissolved in 10 mM sodium phosphate solution. Stereotaxic coordinates targeting the ILA (AP +11.20 mm, ML -0.50 mm and DV -4.40 mm) and ACAd (AP +11.20 mm, …

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

EAU model induction and clinical score

EAU was induced in mice by subcutaneous injection of an emulsion containing 2 mg/mL of retinal antigen interphotoreceptor retinoid-binding protein 1-20 (IRBP1-20) (GiL Biochem, Shanghai, China) and complete Freund’s adjuvant (BD Difco, San Jose, CA, USA), with 2.5 mg of Mycobacterium tuberculosis strain H37Ra (BD Difco, San Jose, CA, USA) in a 1:1 volume ratio. In addition, 0.25 μg of pertussis toxin (PTX) (List Biological Laboratories, Campbell, California, USA) dissolved in PBS was injected on the same day and 2 days after immunization.21,44,45. …

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)

Mice

Rag2−/− (Cat# 033562), Rag2−/−γC−/− (Cat# 014593), Ccl2−/− (Cat# 004434), Foxp3YFP-Cre (Cat# 016959), Scurfy mice (Foxp3−/y, Cat# 006775), Foxp3DTR knock-in mice (Cat# 016958) were purchased from The Jackson Laboratory (Bar Harbour, USA). Ccr2cKO mice were generated by crossing Ccr2loxP (Gempharmatech, Nanjing, China) mice with Foxp3YFP-Cre mice. For Tregs ablation, 6-week-old Foxp3DTR mice were intraperitoneally injected twice a week with 500 ng dose−1 of diphtheria toxin (DT, List Labs, California, USA) for 3 weeks. All mice were housed and maintained under specific pathogen-free (SPF) conditions. …

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

… or lipopolysaccharide (100 ng/mL, List Biological Laboratories, Campbell, CA). …

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://www.listlabs.com/product-information/lipopolysaccharides/

2.1 Reagents and medium

… Pertussis toxin (180) was obtained from List Biological Laboratories. …

2.5 Autoimmune models

To induce experimental autoimmune encephalomyelitis (EAE), female mice were immunized intracutaneously with 200 μg of MOG35-55 peptide emulsified in CFA containing 500 μg of heat-killed Mtb. Mice were intraperitoneally injected with 300 ng of pertussis toxin on day 0 and 2. …

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

2.2. Protocol development steps

The WB samples were collected in sodium heparin tubes (Vacuette, Greiner Bio-one, Kremsmünster, Austria), processed within four hours from blood draw, and prediluted at different ratios in complete medium (M), consisting of RPMI 1640 with GlutaMax with HEPES (Gibco, Life Technology Corporation, Grand Island, NE, USA), 10 % fetal bovine serum (heat inactivated, Sigma Aldrich, St. Louis, MO, USA), and 100 U/mL penicillin/ 100 μg/mL streptomycin (Sigma Aldrich, St. Louis, MO, USA). The prediluted WB was distributed on 48 well tissue culture plates and stimulated with purified genetically detoxified pertussis toxin (PT, 5 μg/mL, heat inactivated at 80 °C for 10 min, LIST Biological Laboratories Inc., Campbell, CA, USA), …

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 and tools table
… Pertussis toxin List Biological Laboratories 180 …

In vivo induction of EAE

Induction and assessment of EAE were performed as previously described with minor modifications (Coll et al, 2015; Shi et al, 2016). Our study exclusively examined female mice because the disease modeled is only relevant in females. Each hind flank of 8–10 weeks female C57BL/6J mice were injected subcutaneously with 250 μg of myelin oligodendrocyte glycoprotein (MOG) peptide 35 to 55 emulsified in complete Freund’s adjuvant containing (0.4 mg/mouse) heat-killed Mycobacterium tuberculosis (4 mg/ml) at two different sites, followed by an intraperitoneal injection of 500 ng of pertussis toxin. On day 2, mice were given a second injection of 500 ng of pertussis toxin. Mice were assigned randomly to two groups and respectively administered with THL (2.5 mg/kg) or vehicle control (DMSO/ PBS) via intraperitoneal every second day starting at the induction of the disease on day 0. …

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