Bacterial Toxin Research Citations

2.11 EAE induction

EAE was induced by subcutaneous injection of 150 μg MOG peptide (amino acids 35‐55), in 0.1 mL of complete Freund’s adjuvant (CFA) emulsion containing 400 μg Mycobacterium tuberculosis (Difco Laboratories, Detroit, MI, USA), into wild‐type or Iqgap1−/− B6 mice on day 0. The mice received intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories, Campbell, CA, USA) on days 0 and 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)

2.9. Experimental Autoimmune Encephalomyelitis (EAE) Induction in a Murine Model

In order to mimic the pathology of human MS, EAE was induced using a murine model as previously described [27,28]. Three-month old C57BL/6J mice (male and female) were immunized with myelin oligodendrocyte glycoprotein (MOG) peptide 35–55 to induce EAE (IACUC #19014). In brief, 300 μg of rodent MOG peptide (amino acids 35–55, New England Peptides, Gardner, MA, USA) in Complete Freund’s Adjuvant (CFA) containing 5 mg/mL killed Mycobacterium tuberculosis (Difco, Thermo Fischer Scientific) was administered into the subcutaneous flank of mice at day 0. At day 0, each mouse received two subcutaneous injections of the MOG solution as well as a 100 µl dose of 2 ng/µl pertussis toxins and virulence factors (List Biological Laboratories Inc., Campbell, CA, USA) diluted in sterile PBS (Thermo Fisher Scientific). Pertussis toxins and virulence factors were administered again on day 2. Pertussis toxins allow an increase in the blood–brain barrier permeability in order to facilitate the incursion of the different treatments into the CNS. …

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)

Additional Pertussis products can be found at https://listlabs.com/products/bacterial-toxins/?sub=pertussis-toxin-and-virulence-factors

… tetanus toxoid (TT) (List Biological Laboratories, Campbell, CA). Peptides were plated at 10 g/ml, TT at 1 g/ml, ECD at 25 ng/ml and …

• Product #191 – Tetanus Toxoid from Clostridium tetani

… Pertussis toxin was obtained from List Biological Laboratories (Campbell, CA) …

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)

2.4. Axon Tracer Injection

An additional group of rats with hemisection SCI (n=8) underwent the same peripheral DCN injections of axon tracers as our previous studies in uninjured normal animals [20]. Three days before the 6-week time point following injury, rats were anesthetized with Ketamine (75 mg/kg)/Dexmedetomidine (0.25 mg/kg). The back skin was shaved, sterilized, and incised at the median line. T7 and T13 DCNs at both sides of animals were identified by counting from the first thoracic DCN branch. DCNs were separated from connective tissues along the lateral dorsal body wall proximally to the latissimus dorsi muscle. Isolectin B4 (IB4, Vector Laboratories, Burlingame, CA) was injected into T7 and T13 DCNs at one side of the animal, and cholera toxin subunit B (CTB, List Biological Laboratories, Campbell, CA) was injected on the other side (Figure 1(a)). Animals with right hemisection injury were divided into two subgroups to inject one label ipsilateral to the injury side (right) and the other to the contralateral side (left). This allowed to determine differences in DCN A and C fiber projections between injured and uninjured sides. A 30-guage needle was inserted 2-3 mm distal to the proximal entry for DCNs at the latissimus dorsi muscle. One microliter of 2% IB4 or CTB dissolved in PBS was injected with a micro syringe (75RN, Hamilton, Reno, NV) driven by a motorized pump (Model #310, Stoelting Co., Wood Dale, IL) at 1 μl/min. The skin incision was stapled, and the animals were recovered from anesthesia with a subcutaneous injection of reversal agent (2 mg/kg, Antisedan).

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

PBMC cultures

The A2Ar agonist, CGS21680 (CGS) (Tocris, Bristol, U.K.) in DMSO (27.7 mg/mL) was diluted in PBS to a final concentration of 0.05 mg/mL. For monocyte assays, α-MSH (Bachem, Vista, CA) at 1 ng/mL and/or CGS (1 μM/well) were added into separate wells. T cells were antigen stimulated with Tetanus Toxoid (TT) (List Biologicals Laboratories, Campbell, CA) at 1 μg/mL was added to cultures. PBMC cultures were maintained in humidified incubator at 37 °C in 5% CO2 for 48 hrs in SFM. At the end of the incubation time, PBMCs were collected by centrifugation (300 g for 5 mins). PBMCs were analyzed with flow cytometry and supernatants were saved for cytokine assays.

• Product #191 – Tetanus Toxoid from Clostridium tetani

Experimental autoimmune encephalomyelitis induction in mice

As described previously (Crocker et al., 2006), experimental autoimmune encephalomyelitis (EAE) was induced in 8-week-old male mice, by subcutaneous (s.c.) administration of 200 μg of MOG35-55 peptide (AnaSpec Inc., AS-60130-10, Fremont, CA, USA) emulsified in complete Freund’s Adjuvant (CFA, Sigma, F5881, Saint Louise, MO, USA) containing 0.2 mg of Mycobacterium tuberculosis H37Ra (Difco Laboratories, #231141 Detroit, MI, USA) into the flanks of both hind-limbs. Control animals received s.c. injections lacking MOG35-55 peptide. At the time of immunization and 48h later, mice further received intraperitoneal (i.p.) injections with 500 ng of pertussis toxin (List Biologicals, #180, Deisenhofen, Germany). …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer

EAE induction

Mice were immunized subcutaneously with 100 μg recombination MOG (rMOG, GQFRVIGPGYPIRALVGDEAELPCRISPGKNATGMEVGWYRSPFSRVVHLYRNGKDQDAEQAPEYRGRTELLKETISEGKVTLRIQNVRFSDEGGYTCFFRDHS YQEEAAMELKVED) peptide or 200 μg MOG35–55 (MEVGWYRSPFSRVVHLYRNGK) peptide emulsified in complete Freund’s adjuvant (Sigma, St. Louis, MO, USA) containing Mycobacterium tuberculosis H37Ra (Difco Laboratories, Detroit, MI, USA) on 0 day and then were injected intravenously with 300 ng pertussis toxin (PT, LIST BIOLOGICAL LABORATORIES, INC.) both immediately after 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)

… Transient depletion of DCs (CD11chigh MHC 25 Class II+ cells) in experimental mice was achieved with administration of Diphtheria toxin (D.tox; 26 8ng/g body weight, List Biological Laboratories Inc. CA). …

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

Experimental autoimmune encephalomyelitis induction and behavioral evaluation of neurologic deficits

Experimental autoimmune encephalomyelitis (EAE) was induced by subcutaneously injecting a solution of myelin oligodendrocyte glycoprotein 35‐55 peptide (200 mg; GenScript, Nanjing, China) emulsified in incomplete Freund adjuvant (Thermo Fisher Scientific) and 500 mg of inactivated Mycobacterium tuberculosis (Thermo Fisher Scientific) into the hind flank of C57BL/6J and TG mice. Two days after immunization, mice were intraperitoneally injected with 200 ng of pertussis toxin (List Biological Laboratories, Campbell, CA, USA).

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)