Bacterial Toxin Research Citations

Discussion:

The proportionality model of intoxication suggests the cytosolic stability of the toxin A chain will directly impact the capacity for recovery from intoxication, which is consistent with our observations for Dtx: as shown in Figs 56, cells could only recover from a minimal initial inhibition of protein synthesis by this relatively stable toxin35. Other reports further suggest the extent of intoxication is linked to the efficiency of toxin clearance from the cytosol. For example, cellular resistance to ricin or EtxA results from enhanced degradation of the cytosolic toxin33,34,36. Conditions that impede the turnover of cytosolic toxin likewise generate cellular sensitivity to ricin or Stx15,32,33,36. These collective observations indicate the extent of intoxication is directly linked to how much toxin is in the cytosol, with the amount of cytosolic toxin representing a balance between toxin delivery to the cytosol and toxin removal from the cytosol.
 

Toxins:

… EtxA and Dtx were purchased from List Biologicals (Campbell, CA). …

• Product #160 – Exotoxin A from Pseudomonas aeruginosa
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

LPS Challenge:

… On the morning of the LPS challenge, we injected each experimental bird intra-abdominally with 1 mg mL1 lipopolysaccharide from E. coli (O55:B5; List Biological Laboratories Inc.) dissolved in PBS. Eight hours post-injection, we collected blood samples from each experimental bird by puncturing the wing vein with a 26-gauge needle. …

NOTE:  At the time this paper was written, Product #203 (5mg – LPS from Escherichia coli O55:B5) was utilized.

Product #203 (5 mg – LPS from Escherichia coli O55:B5) is no longer sold.

Product #203A (2.5 mg – LPS from Escherichia coli O55:B5) is available for purchase.

• Product #203A – LPS from Escherichia coli O55:B5

EAE model:

C57BL/6J mice were immunized subcutaneously with 100 µg MOG₃₅₅₅ (MEVGWYRSPFSRVVHLYRNGK, AnaSpec Inc), in incomplete Freud adjuvant (IFA, Sigma) containing 400 µg of heat-inactivated Mycobacterium tuberculosis H37 RA (Difco) and received two intravenous injections (i.v.) of 350 ng of pertussis toxin (List Biological Laboratories Inc), on day 0 and day 2 after 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)

BoNT/A Heavy Chain Administration and Groups:

Recombinant BoNT-A HC was purchased from List Biological Laboratories Inc. (Campbell, CA, USA).

Intermittent administration of BoNT/A HC applied in the study was done via two routes: (1) local application of the BoNT/A HC directly onto injury site (4 g/L in 16 L saline); and (2) administration of BoNT/A HC via a lumbar intrathecal catheter [46]. After establishing the rat model, 2 g/L of BoNT/A HC were administered every week to the SCI in the BoNT/A HC treatment groups. Animals were divided into the following groups, with six rats in each group: (1) the control (or pseudo surgery) group for which the skin incisions were made, the laminae were cut, and the spinal cords were exposed but not injured; (2) the SCI-only group for which unilateral (left side) lumbar spinal cord injuries were made (to prevent the vehicle intervention on BoNT/A heavy chain, the same volume of sterile saline was applied in SCI-only animal with same method and period); and (3) the SCI with BoNT/A HC treatment, in which BoNT/A HC was administered every week, and the injury was made. This group was further divided into four groups (with six rats each) based on different periods post-injury, i.e., two days, one week, two weeks and four weeks. …

• Product #612A – Botulinum Neurotoxin Type A Heavy Chain, Recombinant, Binding Domain

Toxin A inhibition assays:

Vero cells (CCL-81; ATCC, Manassas, VA) were maintained in complete media (MEM+antibiotic/antimycotic+10% FBS) in T-75 flasks at 37C, 5% CO₂. For TcdA inhibition assays, sterile 96-well tissue culture plates (Thermo-Fisher, Ottawa, ON, Canada) were seeded with ~210⁴ cells/well in a total volume of 200L of complete media. Plates were incubated for 24h at 37C, 5% CO₂. Media was then carefully removed from each well and replaced with 200L of fresh media, 20L of antibody (serially diluted in sterile PBS, giving final in-well concentrations ranging from 1M to 1.95nM) and 10L of TcdA (List Biological Laboratories; final in-well concentration of 10ng/mL corresponding to 32.5pM) for all wells. …

• Product #152C – Toxin A from Clostridium difficile

Cell culture studies:

… For toxin neutralization, we mixed different concentrations of CT (List Biological Laboratories, Campbell, CA) and nanoparticles, incubated for 1 h, and added the mixture to the cell monolayers. After 2 h, supernatants were collected and assayed for cAMP by enzyme immunoassay (Cyclic AMP ELISA Kit, Cayman Chemical Co., Ann Arbor, MI). All cAMP measurements were done without additional acetylation. …

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

In vivo effect of CT in mice

Transgenic mice +/- or -/- for the GalNAcT-gene as described above were fed with 10 μg CT (Azide free, List Laboratories) in PBS with 3% NaHCO3. …

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

Immunofluorescence analysis:

… Consequently, sections were incubated with the following primary antibodies at 4C for 24 h: polyclonal rat anti-Cav3.1, anti-Cav3.2, (1:200 dilution; Alomone Labs, Jerusalem, Israel), Goat anti-CTB (cat. no. 703; List Biological Laboratories, Campbell, CA, USA), …

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

… MAPKKide was obtained from List Biological Laboratories (Campbell, CA 

Author did not specify which List Labs MAPKKide was utilized; the following are available:

• Product #530 – MAPKKide Peptide Substrate (o-Abz/Dnp) for Anthrax Lethal Factor
• Product #531 – MAPKKide Peptide Substrate (DABCYL/FITC) for Anthrax Lethal Factor
• Product #532 – MAPKKide Plus (AMC) Specific Substrate for Anthrax Lethal Factor

EAE:

… Two-hundred nanograms of pertussis toxin (PT; List Laboratories, Campbell, CA, USA) were injected intraperitoneally on days 0 and 2. For passive transfer EAE, donor mice were immunized as describe above. …

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)