Bacterial Toxin Research Citations

EAE immunizations and scoring

8-10-week-old mice were immunized for EAE as previously described [8] with minor modifications. Briefly, mice were administered murine MOG 35-55 (Johns Hopkins Peptide Synthesis Core Facility, Baltimore) in incomplete Freund’s adjuvant (Sigma-Aldrich) supplemented with heat-killed mycobacterium tuberculosis (Sigma-Aldrich) via two subcutaneous flank injections. On days 0 and 2, mice received an intraperitoneal injection of 250 ng pertussis toxin (List Biological Laboratories). …

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 (Escherichia coli O113) is obtained from List Biological Labs, Inc., Campbell, CA, USA …

• Product #433 – LIST™ HPT™ from Escherichia coli O113

4.4. Experimental autoimmune encephalomyelitis (EAE)

To induce a chronic form of EAE, mice were subjected to a protocol in which MOG35–55, a fragment of myelin oligodendrocyte glycoprotein, is twice administered peripherally in adjuvant (Constantinescu et al., 2011). 200 μg of MOG35–55 in a 1:1 emulsion of phosphate-buffered saline (PBS) and Difco Complete Freund’s Adjuvant (BD, Franklin Lakes, NJ, USA) supplemented with an additional 100 mg of Difco Mycobacterium tuberculosis H37RA (BD, Franklin Lakes, NJ, USA) was split into two equal portions and subcutaneously injected in the left and right flanks posterior to the forelimbs. This initial injection was specified as day 0 of EAE. This protocol results in the expansion of T cells autoreactive to myelin. Additionally, the mice received a peritoneal injection of 300 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA) dissolved in PBS. Pertussis toxin causes permeabilization of the blood brain barrier, thus allowing the T cells targeting MOG to enter the CNS. After two days, the mice were given an additional 300 ng booster injection of pertussis toxin to maintain permeabilization of the blood brain barrier. …

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)

Pertussis toxin List Biological Laboratories Cat# 180

Treatment of experimental autoimmune uveitis

Starting two weeks before immunization, B10.RIII mice were fed folate deficient diet and continued for a total of 24 days until euthanasia. To induce EAU, the animals were actively immunized on day 0 with a single subcutaneous injection of 50 μg of human interphotoreceptor retinoid-binding protein (IRBP) peptide 161-180 (IRBP161-180, SGIPYIISYLHPGNTILHVD; GenScript, cat.# RP20268), emulsified in complete Freund’s adjuvant (CFA) containing 2.5 mg/mL of Mycobacterium tuberculosis (BD Difco™). Pertussis toxin (List Biological Laboratories, Campbell, CA), a Th1-biased adjuvant, was given as 2 × 200 ng intraperitoneal doses on days 0 and 2 to enhance organ-specific autoimmunity. …

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

2.3. EAE induction and NPC transplantation

Females 6 to 8 weeks old SJL mice were immunized with 175 μg Proteolipid protein (PLP) 139‐151 peptide diluted in normal saline (0.9% NaCl) and emulsified with complete Freund’s adjuvant containing 5 mg/ml H37RA (Difco Laboratories, Detroit, MI), supplemented with 4 mg/mL of heat-killed Mycobacterium tuberculosis (H37Ra, Difco, Lawrence, KS). Bordetella pertussis toxin (50ug, List Biological Industries) was diluted in 0.2 ml normal saline and injected intra-peritoneally immediately after subcutaneous injection of PLP emulsion. A second bolus of pertussis toxin was administered intra-peritoneally 48 h after primary injection of PLP emulsion. …

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)

2.2 BoNT

rBoNT/A1 gene synthesis, expression in E. coli, and purification methods are described by Hooker et al.15 rBoNT/A1 activity in vitro was confirmed through cell-based assays (SNAP25 cleavage) using rat embryonic spinal cord neuronal (eSCN) cultures. rBoNT/A1 activity ex vivo was shown in the mouse phrenic nerve hemidiaphragm.15

Research-grade purified nBoNT/A1 (150 kDa) was purchased from List Biological Laboratories. The molecular activity was previously confirmed in glycine and glutamate release assays, in SNAP25 cleavage using either rat eSCN or spinal cord neuronal cultures, and in the mouse nerve phrenic hemidiaphragm assay.10, 15 nBoNT/A1 was reconstituted in 1 mg/ml bovine serum albumin (Sigma-Aldrich) in Dulbecco’s phosphate-buffered saline (Sigma) to obtain a 0.1 mg/ml stock solution that was stored at −80°C as single-use aliquots. rBoNT/A1 was stored at −80°C, as single-use aliquots, at 0.1 mg/ml in phosphate-buffered saline (KH2PO4 1 mM, Na2HPO4 3 mM, NaCl 155 mM, pH 7.4). Working solutions were prepared with gelatin phosphate buffer (GPB) to yield the final desired concentrations (see below). GPB was prepared using 0.2% v/v gelatin (Prionex®, Sigma), 0.7% w/v Na2HPO4 (Emsure®, Merck), and sterile water for irrigation (Ecotainer®, Aqua B. Braun). The final pH was adjusted to 6.5 using orthophosphoric acid (85%; Merck).

• Product #130 – Botulinum Neurotoxin Type A from Clostridium botulinum

… EAE was induced with 50 µg myelin oligodendrocyte glycoprotein (MOG35-55; Synpeptide, Shanghai, China) emulsified with 1 mg/ml complete Freund’s adjuvant (CFA; Sigma-Aldrich, Oakville, Ontario, Canada), and 200 or 400 ng of pertussis toxin (PTX; lot #180235A1A, CHO cell assay 0.03 ng/ml; List Biological Laboratories, Campbell, CA), as previously described (Segal et al., 2020) …

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

Pertussis Toxin (PT) was purchased from List Biological Laboratories (Campbell, CA USA)

Induction of EAE. EAE was induced as described (17). In brief, acclimatized, 9-week-old, female C57BL/6J mice were immunized subcutaneously in the flank with 100 μg MOG emulsified 1:1 in IFA supplemented with 4 μg/mL of M. tuberculosis (H37Ra; Difco Laboratories). Animals received 200 ng by i.p. injection of PT on days 0 and 2 postimmunization. R-EAE was induced in acclimatized 9-week-old female SJL/J mice by immunization subcutaneously with 100 μL (50 μg) of PLP emulsified 1:1 in IFA containing 4 μg/mL of M. tuberculosis. …

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)

… Pertussis toxin was from List Biologicals.

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 8–12 weeks old female C57BL/6J WT and ACKR1–/- C57BL/6J mice as described before [59, 60]. Pertussis toxin (List Biological Laboratories, Campbell, US) (300 ng in 100 μl PBS/mouse) was injected intraperitoneally (i.p.) on day 0 (immunization day) and 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)