Bacterial Toxin Research Citations

2.3 MOG35–55 EAE immunization and administration

MOG35-55-induced the EAE in C57BL/6 mice was set up as described previously[20].Briefly, experimental mice were injected subcutaneously on day 0 with 150 µg MOG35-55 peptide emulsified in Freund’s complete adjuvant (CFA) containing
Mycobacterium tuberculosis strain H37Rv (2 mg/mL) (Difco Laboratories, Franklin Lakes, NJ, USA), then mice were injected intravenously with an additional 300 ng of pertussis toxin (PTX) (List Biological Laboratories Inc., Campbell, CA, USA) injection in 100 µl of PBS on day 0 and day 2 following sensitization. …

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)

Transplantation of ex vivo conditionally reprogrammed culture

Conditionally reprogrammed cultures (CRCs) of ASC and AC tumors isolated from KL;CC10 and KL;SPC mice were previously generated and were maintained in co-culture with γ-irradiated NIH3T3 fibroblasts in F12/DMEM media supplemented with 5% HI-FBS, 5 mg/ml insulin (I2643, Sigma-Aldrich), 24 mg/ml adenine (A2786, Sigma-Aldrich), 10 mM Y-27632 (ALX-270-M0055, Enzo Life Sciences), 10 ng/ml human recombinant epidermal growth factor (hr-EGF; 354052, BD Pharmigen), 0.4 mg/ml hydrocortisone (Sigma-Aldrich), 10 ng/ml cholera toxin (100B, List Biological Laboratories) and 1% penicillin/streptomycin (15140122, Thermo Fisher Scientific). 5-10×105 CRC cells were subcutaneously injected to the flanks of nude mice and the tumors were measured weekly with a caliper.

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

Antibodies were obtained from the indicated vendors: … and TcdB (List Bio, 754 A).

EAE

… 200 µg MOG35-55 peptide (Genemed Synthesis) in complete Freund’s adjuvant supplemented with 4 mg/ml Mycobacterium tuberculosis (BD Difco). Four hundred nanogram pertussis toxin (List Biological Laboratories) was administrated intraperitoneally at days 0 and 2 post-immunization. EAE symptoms were scored daily as follows: 0, asymptomatic; …

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)

2.2.1. Induction of EAN

The P0180–199 peptide was synthesized by Ontores Biotech (SSKRGRQTPVLYAMLDHSRS, >98% purity; Hangzhou, China). Freund’s incomplete adjuvant and Heat killed Mycobacterium tuberculosis (strain H37RA) were purchased from BD Difco (#263910, #231141; MI, USA). Pertussis toxin (PTX) was purchased from List Biological Laboratories (#181; CA, USA). Briefly, mice were anesthetized with isoflurane before immunization. PTX solution (1.6 μg/mL, 1.2 μg/mL and 1.2 μg/mL in 250 μL volume each were intraperitoneally administered on Day −1, 1, and 3, respectively. The P0180–199 peptide of 2 mg/mL was emulsified in Freud’s complete adjuvant (20 mg/mL solution of Mycobacterium tuberculosis in Freund’s incomplete adjuvant). The emulsive inoculum of 50 μl was injected subcutaneously into mice on days 0 and 8 [18].

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

… Mice then received an intraperitoneal injection of 180 ng freshly diluted pertussis toxin (List Biological Laboratories or MilliporeSigma) 2 to 5 hours later and again in 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)

Botox injection

Botox (Botulinum Neurotoxin Type A, List Labs #130A) was reconstituted to a stock solution (20 ng/µl) with distilled water containing 1 mg/ml bovine serum albumin. To make injection solution, the stock was diluted with phosphate buffered saline to 10 pg/µl. Mystacial pads were subcutaneously injected with 1 µl of solution via microliter syringe (Hamilton). At first injection, mice stopped whisking within hours, and paralysis from a single injection lasted 7 days with full whisking recovering within 2 weeks. To achieve continuous paralysis across the Botox imaging period, we injected a supplemental 50% dose once per week. We inspected mice daily to confirm the absence of whisking.

Experimental Autoimmune Encephalomyelitis

Active EAE experiments were conducted via immunization of 8-12 week old female C57BL/6 mice with MOG35-55 (Alpha Diagnostic), as previously published12,16,19,20 30 but with one 400ng intraperitoneal injection of pertussis toxin (List Labs) on day 2. Passive EAE experiments were performed as per previously described protocols 12,15,19 32 with the following modifications. Active EAE was first induced via immunization with MOG35-55 and one 400ng intraperitoneal injection of pertussis toxin (List Labs) on day two. Splenocytes 2 were isolated from immunized female mice on day 8, cultured in presence of TH1/TH17 3 polarizing conditions, and adoptively transferred into naïve MCAM KO or WT recipient 4 female mice on day 12. The recipient animals received a 200ng intraperitoneal injection of 5 pertussis toxin (List Labs) on day 14.

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)

BoNT/A

BoNT/A(List Biological Labs, 130B) was injected (25 pg μl−1, i.d., five neighbouring sites injected with 20 µl) into the skin three days and one day before tumour inoculation (defined as prophylactic). BoNT/A (25 pg μl−1; i.d., five neighbouring sites injected with 20 µl) was injected around the tumour one day and three days after the tumour reached a volume of around 200 mm3 (defined as therapeutic) in other groups of C57BL/6J mice.

• Product #130 – Botulinum Neurotoxin Type A from Clostridium botulinum

… Mycobacterium tuberculosis, and was injected i.p. on days 0 and 2 with 400ng pertussis toxin (List Biological Laboratories, Inc. Campbell, CA), as described [16]. …

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)