Bacterial Toxin Research Citations

 Cell culture:

RAW 264.7 cells were maintained in RPMI1640 supplemented with 10% FBS and 200mg/ml gentamicin (Invitrogen Life Technologies). For stimulation of the RAW 264.7 cell, cells were plated in 6-well culture plates (3 106 cells/well) overnight. The overnight growth was performed to allow resting before stimulation. Cells were then washed twice with Opti-MEM (Invitrogen Life Technologies) and stimulated with 50ng/ml E. coli O111:B4 Ultrapure LPS (List Biological Laboratories) …

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

Reagents:

… Pertussis toxin was obtained from 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)

EAE:

… the emulsion was mixed with Freund full adjuvant (Difco Laboratories, USA) containing the death bacterium, and the Pent Balbiter was immunized with the lateral abdominal subcutaneous of the mouse. At the first immunization and after 48 hours, 50 ng whooping toxins per pet (List biological Laboratories, 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)

Disease induction:

Active EAE was induced in mice by subcutaneous injection in the lower flank with 200µg MOG3555 peptide emulsified at a 1:1 ratio with CFA containing 200µg heatkilled Mycobacterium tuberculosis H37RA (Difco, Detroit, MI). Pertussis toxin (PTx; List Biological Laboratories, Campbell, CA) was administered on day 0 (75ng) and day +2 (200ng) with respect to the immunization day. …

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 (PTX, List Biological Laboratories, London, UK) was injected on days 0, 2, and 9 after immunization (300 ng ip). Five mice served as controls and were maintained on a normal diet for the duration of the experiment …

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)

EAE:

… The mice also received an injection of 500ng pertussis toxin (List Biological Labs, Campbell, CA) in 200 µl 0.1M phosphate-buffered saline intraperitoneally immediately after the first immunization with MOG and again 48 hours 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)

… Peptides were also conjugated to cholera toxin B subunit (CTB) (List Biological Labs, Campbell, CA) (peptide:CTB ratio, wt/wt; 2:1) using a two step gluteraldehyde (Sigma-Aldrich) reaction, as described previously (McKenzie and Halsey, 1984; Naz and Chauhan, 2002). …

Vaccination:

… For i.m./s.c. immunizations, each animal received 50 l of total volume of the vaccine containing 50 µg of each peptide conjugated to a carrier protein, emulsified with Freunds adjuvant (Sigma-Aldrich). The first injection was given with Freunds complete adjuvant, while the subsequent booster injections were given with Freunds incomplete adjuvant. For i.n. administration, each animal received 10-15 µl volume of the vaccine per nostril containing a total of 50 µg of each peptide conjugated to CTB. Each animal received a total of 3-4 booster injections. …

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

Neuronal tracing:

Neuronal tracing was carried out in old and young wild type and rd/rd cl mice (n = 3 young wild type, n = 3 young rd/rd cl, n = 2 old wild type and n = 2 old rd/rd cl). Mice were anaesthetized with ketamine hydrochloride, 60 mg kg¹ and xylazine, 7 mg kg¹ and a topical analgesic ophthalmic solution (Proparacaine parachloride). Cholera toxin Subunit B tracer (CTB, List Biological, Campbell, CA) (1% diluted in sterile distilled water) was injected into the vitreous chamber of one eye with the aid of a micropipette (50 mm tip) sealed to the needle of a 5 µL Hamilton syringe. …

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

Antigens and EAE induction:

The mouse MOG₃₅₅₅ peptide (MEVGWYRSPFSRVVHLYRNGK) was synthesized and purity confirmed by HPLC (GenScript, Piscataway, NJ). EAE was induced in mature mice (68 weeks old) by immunization with 100 µg MOG₃₅₅₅ peptide emulsified (1:1) in IFA with 300 µg Mycobacterium tuberculosis, strain H37RA (Bectin Dickinson, FranklinLakes, NJ). Each mouse received 0.05 mL emulsion (s.c.) at four sites. In addition, animals received pertussis toxin (300 ng i.p.; List Laboratories, Campbell, CA) at 0 h and 72 h 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)

Splenocytes:

Individual spleens were obtained and homogenized as previously described [16]. Splenocytes, at a concentration of 110⁶ cells/ml in 24-well plates, were stimulated overnight with the following agonists: E. coli O111:B4 LPS (List Biological Laboratories, Campbell, CA, USA), … Supernatants were collected for cytokine analysis.

• Product #201 – LPS from Escherichia coli O111:B4