Bacterial Toxin Research Citations

Tetanus toxin assay:

A 712bp DNA (biotin-712-thiol) was made in house using PCR from the aforementioned template DNA. The forward primer was modified to incorporate biotin on one end of the DNA and the reverse primer was modified to incorporate a thiol on the other. The thiol modified end of the DNA was utilized to attach it to an anti-Tetanus toxin IgG1 antibody (-TTAb) utilizing SMCC/NHS-ester chemistry. The biotin modified end of the DNA was utilized to attach the DNA-antibody conjugate to Streptavidin coated magnetic beads. A restriction enzyme digest site was engineered at the biotin conjugated end of the DNA to allow for cleavage of the DNA-antibody conjugate from the streptavidin beads after Tetanus toxin (TT) (List Biological Laboratories, Inc. Cat #190B) binding in plasma. …

• Product #190 – Tetanus Toxin from Clostridium tetani

… Anthrax PA was purchased from List Biological Laboratories Inc. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

All toxins were purchased from List Biological Laboratories.

In addition to anthrax toxin [15], diphtheria toxin [30], and Ebola virus29,31, other pathogenic agents SARS coronavirus [31], Venezuelan equine encephalitis virus (VEEV) [32], Rabies virus [33], Junin virus [34], Chikungunya virus [35], and Clostridium difficile toxin B [36] enter the cytoplasm from endosomes and all require the acidification of endosomes. We demonstrated that all of those pathogenic agents are inhibited by AQ in their respective in vitro cellular assays (Table 3 and Supplementary Fig. 4a). We also tested the ability of DEAQ to inhibit pathogenicity of Junin and Chikungunya viruses, and observed that DEAQ inhibits those two viruses with EC50s similar to those of AQ (Supplementary Table 2).

Other pathogenic agents, including cholera toxin [37], … are transported in a retrograde fashion to the endoplasmic reticulum (ER) and retrotranslocated into the cytoplasm by the host ER associated degradation pathway [37,38]. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis
• Product #150 – Diphtheria Toxin, Unnicked, fromCorynebacterium diphtheriae
• Product #155A – Toxin B from Clostridium difficile 

Author did not specify which Cholera toxin was utilized.  List Labs provides Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae and Product #101 – Cholera Toxin from Vibrio cholerae.

Reagents:

… Pertussis toxin was purchased from List Biological Laboratories (Campbell …

Induction and clinical evaluation of EAE:

… On day 0, after anaesthesia, we injected subcutaneously 50 µl of the emulsion in each of the two hind footpads and immediately thereafter 100 ng pertussis toxin in 50 µl saline was intravenously (i.v.) administered in the tail vein. On day 2, a second dose of pertussis toxin was i.v. administered in the tail vein under anesthetized conditions. …

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 and EAE:

Hdac11 constitutive KO mice were generated at the Institut Clinique de la Souris (ICS) in collaboration with Merck & Co. and obtained via Taconic. These mice were backcrossed with the C57BL/6 background for successive generations and then bred to homozygosity. WT C57BL/6 mice were used as controls for these experiments. To induce EAE, WT and HDAC11 KO female mice between 8 and 12 wk of age were immunized with MOG₃₅₅₅ peptide emulsified with CFA (Sigma-Aldrich)-containing Mycobacterium tuberculosis (200 ng/ml; Difco). For each animal, 100 µl of emulsion per site was deposited subcutaneously in the upper and lower backs. In addition, each animal was administered pertussis toxin (islet-activating protein, 500 ng, i.p.; List Biological Laboratories) at the time of immunization and again 1 d 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)

… Pertussis toxin (200 ng, List Biologicals) was injected i.p. on day 0 and day 1 after MOG 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)

… Diphtheria toxin (DT; 50 g/kg body wt; List Laboratories, Campbell, CA) was administered once (in the absence of UUO) or at stated times after UUO via ip doses …

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

… 0.2 µl of CTb (10 µg/ml, List Biological Laboratories, CA, USA) was injected into the vHPC, …

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

… Cholera Toxin from Vibrio cholerae, List Biological Labs, Cat# 100B. Binary Toxin from Clostridium difficile, Subunit A, List Biological Labs, Cat# 157A. Binary Toxin from Clostridium difficile, Subunit B, List Biological …

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

Induction of Experimental Autoimmune Encephalomyelitis (EAE) in female C57BL/6 mice:

Experimental autoimmune encephalomyelitis (EAE), a frequently used mouse model of multiple sclerosis (Robinson et al, 2014), was induced in female C57BL/6 mice largely as previously described (Williams et al, 2011). For EAE induction, female C57BL/6 mice older than 10 weeks (body weight between 20 and 25 g) were selected. Mice were injected subcutaneously into the axilla and groin with a total of 200 g of the encephalitogenic MOG₃₅₅₅ peptide of mouse myelin oligodendrocyte glycoprotein (MEVGWYRSPFSRVVHLYRNGK; > 90% purity, synthesized by Dr. Martin Jung, Institute of Medical Biochemistry and Molecular Biology, Medical School Homburg, Saarland University) in sterile water (2 mg/ml) emulsified with an equal volume of complete Freund adjuvant (CFA), consisting of incomplete Freund adjuvant (iCFA, Sigma; #F5506) to which 10 mg/ml inactivated Mycobacterium tuberculosis were added (Fisher Scientific #10218823), as previously described (Williams et al, 2011). 200 ng of pertussis toxin (PTX) from Bordetella pertussis (List Biological Laboratories Inc. #181, via Biotrend, Cologne, Germany) in a volume of 100 l sterile H₂O was injected intraperitoneally on the same day (day 0, 12 h after MOG₃₅₅₅ peptide injection) and also on the subsequent day (day 1, 1620 h after first PTX injection). …

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