Bacterial Toxin Research Citations

In vivo cell depletions

Animals were depleted of CD4+ or CD8+ T cells as previously described [19,85]. Briefly, animals were treated with 0.5 mg of rat anti-CD4 mAb GK1.5 (Leinco) or 0.2 mg rat anti-CD8 mAb 2.43 (Leinco) in 200 μl of DPBS i.p. one day prior to challenge. Treatment was repeated once weekly for the entirety of each study. For depletion of B cells, animals were treated with 250 μg of rat anti-CD20 mAB MB20-11 (BioXcell) in 200μl of DPBS i.p. one week prior to challenge [86]. Anti-CD20 treatment was repeated on day 14 post infection. Control animals for both T and B cell depletions received equivalent dosages of rat or mouse IgG (Leinco or Southern Biotech) respectively. Upon euthanasia, depletion of splenic CD4+ T, CD8+ T, or B cell populations were confirmed to be ≥ 90% effective via flow cytometry. TReg were depleted in DEREG animals by administering 1 μg DTX (List Biological Laboratories) resuspended in 100 μL DPBS i.p. to each animal on day 14 and 15 post challenge [59]. WT animals were administered an identical dose of DTX as a control. Blood was collected from animals to confirm systemic depletion of FoxP3+CD4+ T cells seven days post treatment (S3A Fig).

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

2.2. EAE induction and treatment
EAE was induced according to previously established methods [17,18]. Briefly, mice were subcutaneously immunized with 200 mg MOG 35–55 peptide emulsified in complete Freund’s adjuvant containing 0.4 mg Mycobacterium tuberculosis H37 Ra (BD Difco) and intraperitoneally injected with 200 ng pertussis toxin (List Biological Laboratories) on days 0 and 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)

Tracer injection

Rats received a subcutaneous injection of the nonsteroidal anti-inflammatory ketoprofen
(2.5mg/kg, s.c.). and were anesthetized with isoflurane (5% induction, 2.5% maintenance) in
O2 before being placed in a stereotaxic frame. Animals were placed on a heating pad for the
entire procedure. Lidocaine (1mg/kg) was injected locally before incision of the skin. Then,
craniotomies were performed above the brain areas of interest. A retrograde tracer cholera
toxin subunit B (CTB, 0.5 μl, List Labs, Campbell, CA, USA) was injected into the right NAcC
[anteroposterior (AP), +1.1mm for males and +1.2mm for females from bregma; lateral (L),
2.2mm from the midline; dorsal (D), -7.4mm from the skull surface]. Following injection, the
incision was sutured, and the rats were placed in their homecage for 2 weeks before brain
collection.

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

EAE

EAE was induced in 8–12 weeks old female C57BL/6J mice as previously described (17, 70). … On day 0, mice were immunized by injecting 50 μl of the emulsion subcutaneously (SC) into each of two sites on the lateral abdomen. On days 0 and 2, the mice were injected IP with 250 ng pertussis toxin (List Biologicals, catalog no. 181) dissolved in PBS. On day 7, they were weighed and began scoring.

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

Plasma antibody measurements

Pertussis antigen-specific antibody responses were quantified in human plasma by performing an indirect serological assay with xMAP Microspheres (details described in xMAP Cookbook, Luminex 5th edition). Pertussis, Tetanus, and Diphtheria antigens (PT, PRN, Fim2/3, TT, and DT (all from 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)

• Product #186 – Fimbriae 2/3 from B. pertussis

• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

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

• Product #190 – Tetanus Toxin from Clostridium tetani

Induction and assessment of EAE

Mice were subcutaneously injected with 200 mg MOG (myelin oligodendrocyte glycoprotein) 35–55 (MEVGWYRSPFSPVVHLYRNGKCOOH, eurofins) peptide emulsified in complete Freund’s adjuvant containing 400 mg of Mycobacterium tuberculosis H37Ra (Chondrex), in addition to two intraperitoneal injections of 200 ng of pertussis toxin (List Laboratories) on days 0 and 3. …

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)

4.1. Cell Line and Reagents for Development of the AP-1 Reporter Assay
Murine hepatoma Hepa1c1c7 cells (CRL-2026™) purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) were cultured in an alpha minimum essential medium (MEM), supplemented with 10% FBS, 100 IU/mL penicillin, 100 µg/mL streptomycin, and 2 mM L-glutamine. Cultures were maintained at 37 °C in an incubator with 5% CO2. Lyophilized recombinant protective antigen (PA) and lethal factor (LF) were purchased from List Biological Laboratories, Inc. (Campbell, CA, USA), and reconstituted in sterile water to make stock solutions with a final concentration of 1 mg/mL for LF and 2 mg/mL for PA. The stock solutions were then diluted 10 times with 50% glycerol in sterile water to make stock solutions with a final concentration of 0.1 mg/mL for LF and 0.2 mg/mL for PA. …

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

Author did not specify which Lethal Factor (LF) was utilized; List Labs provides the following LF:

• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis
• Product #169L – Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence

… Pertussis toxin (200 ng/mouse, List Biological Laboratories, Inc., Campbell, CA, USA) in sterile saline was administered via the tail vein on day 0 and day 2 after MOG35-55 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)

CT intoxication of human monolayers

… After 3 days, the monolayers were challenged with 2 μg mL−1 CT (List Labs) on the apical side. …

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

Limbus processing procedures

… 0.1 nM cholera toxin (List Biological Laboratories, Campbell, California, USA), …

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