Bacterial Toxin Research Citations

Disease Induction and Assessment of EAE:

EAE was induced in female C57BL/6 mice (Jackson Laboratories, Bar Harbor, ME), aged 7 to 8 weeks, by injecting subcutaneously 50 µg of myelin oligodendrocyte glycoprotein (MOG)₃₅₅₅ in complete Freund’s adjuvant (Fisher Scientific, Waltham, MA) supplemented with 4 mg/mL Mycobacterium tuberculosis on day 0.^32,33 Intraperitoneal pertussis toxin (300 ng/200 µL; List Biological Laboratories, Hornby, ON, Canada) was administered on days 0 and 2. Laquinimod (25 mg/kg body weight) was given every day by oral gavage in 100 µL of saline, whereas 100 µL of saline was used as the vehicle control. …

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)

Induction of EAE:

EAE was induced in 6-wk-old male mice by s.c. injection with 100 mg myelin oligodendrocyte glycoprotein (MOG)3555 peptide (MEVGWYRSPFSRVVHLYRNGK; United Peptide, Rockville, MD) emulsied in 100 ml CFA (Sigma-Aldrich, St. Louis, MO) containing 2 mg/ml heatkilled Mycobacterium tuberculosis H37RA (Difco, Detroit, MI). Mice were also injected i.p. with 200 ng pertussis toxin (List Biological Laboratories, Campbell, CA) on day 0 and day 2. Clinical signs of EAE were recorded daily using a 04 scoring system: 0, normal; 1, limp tail; 2, unsteady gait; 3, hind-limb paralysis; 4, fore-limb paralysis.

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 1: identification of refeeding-activated neurons in the PVN that project to the dorsal vagal complex

Rats were anesthetized ip with ketamine/xylazine (ketamine 50 mg/kg, xylazine 10 mg/kg body weight) and their heads positioned in a stereotaxic apparatus with bregma and lambda in the horizontal plane. Through a burr hole in the skull, a glass micropipette (20 µm outer tip diameter) containing 0.5% solution of the retrograde tracer, cholera toxin- subunit (CTB; List Biological Laboratories, Campbell, CA), was lowered into the brain at stereotaxic coordinates corresponding to the NTS based on the atlas of Paxinos and Watson (7): 5.0 mm posterior and 1.9 mm dorsal to the interaural line and 0.5 mm lateral to the midline. …

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

Materials:

Recombinant LF and PA were from List Biological Laboratories. …

SDS-PAGE-Based Cleavage Assay:

LF (25 nM in assay buffer) was mixed with compound at the indicated concentration, and the reaction was initiated by adding unlabeled MKK6 to 1 M and stopped after 30 min at RT by adding 4 SDS-PAGE loading buffer. …

Cytotoxicity Assay:

RAW264.7 cells were plated in 96-well dishes at 4  10⁵ cells per well and were allowed to recover for 16 hr, after which the medium was removed and replaced with fresh complete medium (100 l per well) containing 400 M compound or 4% DMSO vehicle control. After 30 min, PA (0.5 g/ml) and/or LF (0.0010.3 g/ml) were added and incubation continued for an additional 4 hr.

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 #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

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

Bacterial Superantigens and Endotoxin:

… Lipopolysaccharide (LPS; Escherichia coli 055:B5) was purchased from List Biological Laboratories, Inc. (Campbell, CA) and reconstituted with PBS prior to use. …

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

Induction and evaluation of EAU:

SLAT/Def6 KO mice and their C57BL/6 controls were immunized with bovine IRBP (150 µg), emulsified in complete Freunds adjuvant (CFA), administered subcutaneously [10]. In addition, the mice were injected intraperitoneally with 0.2 µg pertussis toxin (List Biological Laboratories, Inc., Campbell, CA). The development of ocular inflammation was determined by fundoscopy on day 12 post-immunization and by histological examination on day 14, following euthanization. Severity of disease, on a scale of 04, in half point increments, was scored as detailed elsewhere [11,12].

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)

Reagents:

… Ultrapure lipopolysaccharide (LPS) (Escherichia coli 0111:B4, TLR4 lig- and) was purchased from List Biological Laboratories, Inc. (Vandell Way, CA, USA). …

Results – DNA Stimulates Factor B Synthesis and Release by Macrophages:

Early upregulation of factor B occurs after ischemic tissue injury. However, the endogenous molecules that stimulate factor B production in the setting of ischemic tissue injury have not been identified. We have previously shown that microbial ligands, including LPS and polyinosinepolycytidylic acid (poly I:C) stimulate dynamic synthesis and release of factor B by macrophages (26). We next sought to test the hypothesis that one of several known endogenous mediators of inflammation known to activate patternrecognition receptors would stimulate factor B synthesis. Because we previously found that activation of TLR4 signaling by LPS resulted in synthesis of factor B in macrophages, we therefore studied the effect of several endogenous molecules that have been identified as activators of TLR4 signaling (3335). As previously observed, stimulation of RAW264.7 macrophages with LPS resulted in upregulation of factor B mRNA (9.9-fold, p<0. 001). However, no significant upregulation of factor B was observed after stimulation with fibrinogen, heparan sulfate or HMGB1 at known stimulatory concentrations (Figure 1). …

• Product #421 – ULTRA PURE LPS from Escherichia coli O111:B4

EAE induction:

… CD137L^-/- mice were a gift from Amgen and bred in-house under pathogen-free conditions. Mice were injected subcutaneously with 100 µg of myelin oligodendrocyte glycoprotein peptide fragment 35-55 (MOG_35-55) (Sigma-Aldrich) and 1 mg heat-killed Mycobacterium tuberculosis H37RA (Difco) emulsified in complete Freunds adjuvant. Pertussis toxin (200 ng in PBS; List Biological Laboratories) was injected intraperitoneally on days 0 and 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)

… cholera toxin was from List Laboratories; …

Intragastric antigen sensitization and treatment:

Sensitization was performed by 20 [micro]g OVA gavage with 10 [micro]g cholera toxin in 0.2 ml saline for 5 times once a week using a stainless steel blunt feeding needle. Mice in the non-sensitized control group received saline. Experimental diet included 0.1% or 0.3% red ginseng or fermented red ginseng. Mice were fed the experimental diet for 8 weeks starting at 1 week before the initial sensitization. …

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

EAE induction and clinical evaluation:

EAE was induced by s.c. immunization with 100 mg MOG3555 peptide in 100 ml CFA containing 0.4 mg Mycobacterium tuberculosis H37RA (Difco); 200 ng pertussis toxin (List Biological Laboratories) was administered i.v. on days 0 and 2. Clinical scoring was: 1 = limp tail; 2 = hind limb paresis or partial paralysis; 3 = total hind limb paralysis; 4 = hind limb paralysis and body or front limb paresis or paralysis; 5 = moribund.

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)