Bacterial Toxin Research Citations

In vivo treatments with chemicals and antibodies

For DT-induced depletion of IM, IMDTR mice were injected i.p. with a single dose of 50 ng DT (List Biological Labs, 150), unless otherwise stated. …

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

CD115+ M-MDSC depletion

MMDTR mice were implanted with 2.5 × 105 EL4 cells. Each mouse received one intraperitoneal injection (10 ng/g of body weight) of diphtheria toxin (DT) (List Biological Laboratories, Campbell, CA, USA) on days 7, 9, and 11 after tumor implantation (Supplementary Fig. 7b). Time intervals (every 48 h) of DT injection were optimized by flow cytometry blood analysis (Supplementary Fig. 7a). After sacrifice on day 14 after tumor implantation, single cells were prepared for flow cytometry analysis. CD11b+Ly6G+Ly6Clo cells from naive or TB mice were sorted for OLFM4 immunohistochemistry (IHC) and qRT-PCR analysis. C57BL/6 mice were used as a negative control to exclude effects related to DT.

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

2.2.1. Antigens and Couplings

Tetanus toxoid was purchased from Massachusetts Biological Laboratories (Boston, MA, USA) and diphtheria toxoid was purchased from List Biological Laboratories (Campbell, CA, USA). Antigens were coupled to MagPlex microspheres (Luminex Corp, Austin, TX, USA), as described previously [5,23], using antigen concentrations of 12.5 µg/12.5 × 106 microspheres for tetanus toxoid and 60 µg/12.5 × 106 microspheres for diphtheria toxoid.

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

… Anthrax Protective Antigen List Biological Labs Cat# 171E Anthrax Lethal Factor List Biological Labs Cat# 172C …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

2.2. EAE Induction and Scoring

Female mice, aged 6–8 weeks, were immunized subcutaneously with 50 mg MOG35–55 peptide (synthesized by Toray Research Center, Japan) containing 1 mg heat-killed Mycobacterium tuberculosis H37RA emulsified in complete Freund’s adjuvant (Difco, Franklin Lakes, NJ, USA). The mice were injected intraperitoneally (i.p.) with 100 ng of Pertussis toxin (List Biological Laboratories, Campbell, CA, USA) at the time of immunization and 2 days later. …

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)

… Three wild-type mice received stereotaxic injections of FG (2.0%, Fluoro-Gold; Fluorochrome) in the LC and CTB (0.5% Cholera Toxin B subunit, LIST Biological Laboratories) in the PVT. Brain tissues were collected 7 d after surgery and processed for histology. Antibodies against CTB (1:500 dilution; 703/AB_2314252, LIST Biological Laboratories) and FG (1:50 dilution; Fluorochrome) were applied along with anti-TH to identify …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

EAE induction

Mice were induced with active EAE six to ten weeks post Tamoxifen treatment (after replenishment of IGF-1R+ blood monocytes by natural turnover), as previously described [44]. Briefly, at day 0, mice were immunized with an emulsion of 200 μg MOG peptide (MOG35-55, Genscript, USA) and complete Freund´s adjuvant (Santa Cruz Biotechnology) containing Mycobacterium tuberculosis (Difco) (100 μl/mouse subcutaneously, in the left and right flank and at the tail base). Furthermore, 400 ng pertussis toxin (List Biological Laboratories, Campbell, CA, USA) diluted in sterile DPBS was administered intraperitoneally at day 0 and 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)

PC-LCA-HPLC

BoNT/A light chain activity was measured after toxin was recovered from reconstituted product using an established PC-LCA-HPLC method. … After washing, digestion buffer (200 μL) containing 0.5 mM zinc acetate, 0.5% Triton X-100 (Dow Chemical Company, Midland, MI, USA), 0.5 mg/mL human serum albumin (HSA), and 2 mM dithiothreitol (DTT) in 50 mM 4-(2-hydroxyethyl)-1-piperazine ethanesulfonic acid (pH 7.4) was added to each well. All concentrations of Triton X-100, HSA, and DTT were optimized to increase assay sensitivity. Plates were then incubated for 30 minutes at 37°C to facilitate toxin reduction. Aliquots (50 μL) of 200 μM SNAPtide 520 substrate (List Biological Laboratories Inc., Campbell, CA, USA) were added to each well, then incubated at room temperature for 72 hours to allow for substrate cleavage. …

• Product #520 – SNAPtide® Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin

Bphs phenotyping

Bphs phenotyping was carried out as previously described15. Briefly, mice were injected with purified PTX (List Biological Laboratories, Inc.) in 0.025 M Tris buffer containing 0.5 M NaCl and 0.017% Triton X-100, pH 7.6. Control animals received carrier alone. Three days later, mice were challenged by injection with histamine (milligrams per kilogram of body weight [dry weight], free base) suspended in phosphate-buffered saline (PBS). Deaths were recorded at 30 min post-challenge. The results are expressed as the number of animals dead over the number of animals studied.

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)

… After the last day of fentanyl self-administration training, we injected CTb (List Biological Laboratories) into Pir. We injected 40 nl of 1% CTb unilaterally into Pir with the needle left in place for an additional 5 min (Mahler and Aston-Jones, 2012; Marchant et al., 2014, 2016; Venniro et al., 2017b). We counterbalanced injections into either the left or right hemisphere and used a 1.0ml, 32 gauge Neuros Syringe (Hamilton) attached to UltraMicroPump III with SYS-Micro4 controller (World Precision Instruments). The coordinates for Pir were AP, 13.4 mm; ML, 64.3 mm (10° angle lateral to midline); DV, 6.7 mm. These coordinates are based on pilot studies to optimize spread of CTb within Pir and minimize spread to adjacent areas. …

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