Bacterial Toxin Research Citations

Enzyme-linked immunosorbent assay(ELISA)

… To measure the levels of IgG against CCP2, we used a commercially available ELISA
kit (Euro-Diagnostica, Malmö, Sweden). For the detection of anti-inuenza, and anti-diphtheria toxin
antibodies, 96-well plates were coated with 1 μg/ml of A/H1N1 subunit (Bio-Rad, Hercules, CA, USA) and
diphtheria toxin (List Biological Laboratories, Campbell, CA, USA), respectively. …

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

Reagents and cell lines.

… 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)

… Mice also received 200 ng of Bordetella pertussis toxin (List Biological Laboratories) intraperitoneally 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)

Experiment 3:

… Upon examining the sections through the PBel in animals with injections of AAV-FLEX-ArchT-GFP into the DR of Sert-Cre mice, we found intense innervation of the external lateral part of the parabrachial (PBel) area. To further confirm that PBel receives innervation from serotonergic neurons in the DR, we also injected Sert-L10 mice (n = 4) with retrograde tracer, cholera toxin subunit b (24 nl, 0.2% – CTb, List Biological Laboratories Inc.) unilaterally in the lateral PB. One week later these mice were sacrificed, brains perfused, and tissue sections processed for immunohistochemistry for CTb using the procedure described below.

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

… C. difficile Toxin B (TcdB) List Biological Laboratories Cat#155A …

• Product #155 – Toxin B from Clostridium difficile

… Free-floating sections were washed in 0.01 M PBS, followed by incubation in hydrogen peroxide for 20 min and incubated for 24 h at room temperature (RT) with a goat anti-CT-B antibody (1:10,000 List Biological Laboratories, CA, USA) …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

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

IFNγ ELISPOT assay

Peripheral blood mononuclear cells (PBMCs) were collected using mononuclear cell preparation tubes (CPT) prior to treatment initiation and at 3 and 6 weeks and were cryopreserved in liquid nitrogen at 5×106 cells per vial in cryopreservation media containing 10% dimethyl sulfoxide (DMSO). The procedure for the FRα ELISPOT assay has been described previously.22 Briefly, 2.5×105 PBMCs/well were plated in triplicate into 96-well round bottom plates in 200 µL of RPMI1640 containing L-glutamine, penicillin, streptomycin, and 10% serum (T cell medium), along with cyclin D1 (control) peptide (MELLLVNKLKWNLAA), FR30, FR56, FR76, FR113, FR238, FRα protein (Sino Biological), tetanus toxoid (List Biological Laboratories), …

• Product #191 – Tetanus Toxoid from Clostridium tetani

Safety assessment in mice and RMs

For preliminary safety studies, 8-week-old female BALB/c mice were obtained from the Jackson laboratory (Bar Harbor, ME) and immunized by the intraperitoneal route with a 40-μg dose of H2O2-CampyC. In a separate control study, mice were treated with 10 μg of E. coli LPS (O111:B4, List Biological Laboratories, Campbell, CA). Animal weights were followed for up to 1 week after immunization. The second phase of our safety assessment was conducted in RM. Two adult females received an IM vaccination with the H2O2-CampyC vaccine, and two adult females received a mock, alum-only IM vaccination. Health metrics, including weight, attitude, appetite, urine, stool, temperature, and vaccine site, were monitored daily for 14 days.

• Product #201 – LPS from Escherichia coli O111:B4

BoNT administration

Research-grade, purified, native BoNT serotype A1 (BoNT-A; 150 kDa) was purchased from List Biological Laboratories (Campbell, CA, USA). It was reconstituted in 1 mg/ml phosphate buffer saline/bovine serum albumin (0.1%) to obtain a stock solution, which was aliquoted and kept frozen at −80 °C. Dilutions of aliquots were performed with 0.2% gelatin phosphate buffer (GPB) to obtain the final concentrations (see below). …