Bacterial Toxin Research Citations

… SF were stimulated with human recombinant TNF … lipopolysaccaride (LPS) from Escherichia coli J5 (100 ng/ml; List Biological Laboratories), …

• Product #301 – LPS from Escherichia coli J5 (Rc)

… Where indicated, Foxp3+ Tregs were ablated in host mice via ip administration of diphtheria toxin (List Biologicals) at 50 mg/kg on the day of adoptive transfer …

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

Mice, peanut sensitization, and EFB-2 treatment

Six-week-old female C3H/HeJ mice purchased from the Jackson Laboratory (Bar Harbor, ME) were maintained in pathogen-free facilities at the Mount Sinai vivarium according to standard guidelines (57). Mice were intragastrically (i.g.) sensitized with 10 mg of homogenized peanut (PN) in 0.5 mL PBS containing 75 mg sodium bicarbonate, 10 µg of the mucosal adjuvant cholera toxin (CT) (List Laboratories, Campbell, CA), and 16.5 µL (1.1 µL/g body weight) of 80 proof Stolichnaya Vodka® (a source of food grade ethanol) to neutralize stomach pH and increase gastrointestinal permeability, three times during week 0 (50). Thereafter, sensitization was done weekly as above except that the CT dose given was 20 µg.

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

… Pertussis toxin (PTX) List Labs Cat# 180 …

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

Antigens for in vitro stimulation

Pertussis toxin (PT) and filamentous hemagglutinin (FHA), the two Bp antigens present in all aP vaccines (36), were selected for in vitro stimulation of blood cells in the Bp-specific whole blood intracellular staining (BpWB-ICS) assay to expand vaccine-induced Bp-specific precursor T lymphocytes. A Bp lysate (BPL) was also used to detect cellular immune responses to Bp antigens that are not included in aP vaccines. A genetically detoxified PT mutant, with an inactivated S1 subunit (R9K, E129A), was chosen for the in vitro T cell assay (LIST Biological laboratories Inc., Campbell, CA, USA, #184) to avoid the toxic activity on target cells (37). …

• Product #184 – Pertussis Toxin Mutant

EAU induction and clinical evaluation

Mice were immunized subcutaneously on day 0 with 300 µg of Interphotoreceptor Retinoid-Binding Protein (IRBP) 651–670 (Hanheng Group, Shanghai, China) emulsified in CFA (incomplete Freund’s adjuvant plus 4 mg/ml of Mycobacterium tuberculosis strain 37RA, Difco). Pertussis toxin (0.5 µg in 200 µl saline; List Biological Laboratories, USA) was administered twice intraperitoneally on day 0 and day 1 after immunization. Disease status was assessed by fundus examination on a scale of 0–4 based on the number, type, and size of lesions and extent of inflammation, as described previously [26].

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)

4.1. Animal Experiments

We purchased six-week-old female C57BL/6 mice from CLEA Japan, Inc. (Tokyo, Japan). …

We divided mice into two groups: the naïve and CFA-injected groups (eight mice per group). For the CFA-injected group, we sensitized mice subcutaneously with CFA that consisted of IFA (BD, Franklin Lakes, NJ, USA) and M. tuberculosis H37 Ra (BD) on days 0 and 19. The final concentration of M. tuberculosis in the CFA emulsions was 2 mg/mL (400 μg/mouse). We also injected CFA-injected mice with 300 ng of PT (List Biological Laboratories, Campbell, CA, USA) intraperitoneally on days 0 and 2. PT has been used by other research groups, including Johanson II et al. [25], for CFA-induced autoimmune models [64]. …

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)

Experimental autoimmune encephalomyelitis

EAE was induced in both male and female mice between 8 and 12 weeks of age. MOG35-55 peptide (100 μg, CSBio; CS0681) was emulsified in complete Freund’s adjuvant containing Mycobacterium tuberculosis (1 mg/mL, Sigma; F5881; M. tuberculosis (BD 231141) added to final concentration of 4 mg/mL) and was injected subcutaneously (100 μL volume) at the base of the tail. Pertussis toxin (200 ng, List Biologicals; 180) was administered i.p. on the day of and 1 day after MOG immunization. …

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

EAE induction and clinical scoring

EAE/optic neuritis was induced with MOG (MOG33-55, GenScript, cat. no.163913-87-9) peptide and PTX (List Biological Laboratories, Campbell, CA, USA, cat. no. 181) in 9-week-old female mice with a procedure modified from the established protocol.29,30,44 Briefly, 225 μg MOG33-55 peptide emulsified with incomplete Freund’s adjuvant (IFA; BD Difco, 263910) and 2.5 mg/mL mycobacterium tuberculosis (BD Difco, 231141) were injected subcutaneously for immunization twice at days 0 and 7, and 500 ng PTX in PBS (2.5 μg/mL) was intraperitoneally injected twice at days 0 and 2 to break down the blood-brain barrier. Mice injected with the same volume of Freund’s adjuvant emulsion without MOG33-55 were used as sham controls. …

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

Generation of hTEC cultures

Cell cultures of primary human TECs (hTECs) were established following the procedure originally described by Green et al33 with modifications. … To enrich for TECs, epithelial cell adhesion molecule (EpCAM)-positive cells were enriched by using anti-EpCAM (CD326) (clone HEA-125, Miltenyi, Bergisch Gladbach, Germany) staining and anti-APC microbeads (Miltenyi) separation. The TEC-enriched cell fraction was plated (2.5 × 104 cells/cm2) on a feeder layer of lethally irradiated 3T3-J2 murine fibroblasts (Kerafast, Boston, Mass) and cultured in a humidified atmosphere of 5% CO2 in growth medium composed of a mixture of Dulbecco modified Eagle medium and Ham’s F-12 (a 3:1 mixture), 10% FCS (ThermoFisher Scientific), insulin (5 μg/mL, Eli Lilly, Indianapolis, Ind), adenine (0.18 mM, Sigma, St Louis, Mo), hydrocortisone (0.4 μg/mL, Sigma), cholera toxin (0.1 nM, List Labs, Campbell, Calif), …

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