Bacterial Toxin Research Citations

2.2. Immunization and collection of specimens

Mice were anesthetized with isoflurane before receiving the vaccine preparation for intranasal immunization. A 20-µl phosphate-buffered saline (PBS) solution containing the Mtb antigen ESAT-6 (5 µg; MyBioSource; MBS204541) in the presence or absence of S100A4 (10 µg; Gentaur Molecular Products; 01-2081A4M; with His-tag) was administered dropwise to the external nares of the mice (10 µl per nostril). Some mice were immunized with ESAT-6 (5 µg) admixed to cholera toxin (CT; 1 μg; List Biological Labs; 100B) as a control adjuvant. Ten days after the last intranasal immunization, various tissues and samples were collected and analyzed, including blood, bronchoalveolar lavage fluid (BALF), nasal lavage, lungs, and spleen.

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

STAR+METHODS
KEY RESOURCES TABLE

CTB List Biological Laboratories Cat: 103B
CT List Biological Laboratories Cat: 100B

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae

Induction of EAU model

On the initial day of the experiment, the mice were administered a subcutaneous injection containing a 1:1 combination of 200 μg of human interphotoreceptor retinoid-binding protein 1–20 (IRBP1–20, GPTHLFQPSLVLDMAKVLLD; GiL Biochem, Shanghai, China) and complete Freund’s adjuvant (Difco, Detroit, MI, USA). The complete Freund’s adjuvant consisted of 2.5 mg/mL of Mycobacterium tuberculosis (BD Difco, San Jose, CA, USA). On the same day of immunization and two days thereafter, an intraperitoneal injection of 0.25 μg pertussis toxin (PTX) diluted in phosphate-buffered saline (PBS) was administered. The PTX used in this study was obtained from List Biological Laboratories located in Campbell, California, 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)

Induction of MOG 35-55 peptide-induced EAE

On Day 0, ~12-week-old female mice were immunized subcutaneously with 200 µg of MOG 35-55 peptide (MEVGWYRSPFSRVVHLYRNGK)118 and 400 µg of Mycobacterium tuberculosis H37Ra (MTb, Difco, Detroit, MI). Incomplete Freund’s Adjuvant (IFA; Gibco, Grand Island, NY) and dried, heat-killed MTb were emulsified with an equal volume of MOG 35-55 dissolved in PBS using a VirTishear Cyclone homogenizer with a 10 mm micro shaft impeller (VirTis, Gardiner, NY). The mice were immunized with 150 µl of the MOG-containing emulsion divided among three subcutaneous sites on the flank (between the shoulder blades and over each hip). Additionally, on Days 0 and 2, the mice were injected intraperitoneally with 200 ng pertussis toxin (PTx) (List Biological Laboratories, Campbell, CA) in 200 µl PBS119.

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)

Rabies virus tracing

… We performed a craniotomy to expose either (1) the ventral portion of the occipital cortex and the lateral aspects of the posterior cerebellum or (2) the lateral prefrontal cortex. We resected the dura and used a Hamiton syringe (30-gauge needle) to place multiple injection tracks into either the cerebellar hemisphere (Crus IIp) or the PrePMd. We injected small amounts (0.2 µl) of a mixture of rabies virus (CVS-N2c 4.5 × 109 pfu/mL; provided by M. Schnell) and 0.02% CTb (List Biological Laboratories) at depths at least 0.5 mm apart along the injection tracks. …

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

… For the in vitro stimulations, PBMCs were seeded at a concentration of 2×105 cells/well in triplicates and stimulated with LPS (8.33 ng/ml, List Biological Laboratories Inc., Campbell, USA) with/without α7nAChR agonist PHA568487 (83 µM) for 4 hours as previously described. Supernatants were analyzed using the Bio-Plex Pro Human Cytokine Panel 17-plex assay (Bio Rad Laboratories, Inc. Hercules, USA) following the manufacturer’s protocol. Inclusion criteria for each participant: Following LPS stimulation, …

Author did not specify which List Labs LPS product was utilized in their research.
List Labs provides the following LPS products: https://www.listlabs.com/product-information/lipopolysaccharides/

MATERIALS AND METHODS

Antigens and standards PT, PRN, and FIM 2/3 were 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)

• Product #186 – Fimbriae 2/3 from B. pertussis
• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

2.3.3. ELISpot Assays

… In brief, ELISpot assay plates (MabTech Inc., Cincinnati, OH, USA) specific for the detection of primate IFN γ were used according to the manufacturer’s instructions. Diluted PBMC cells were dispensed (100 µL/well) into a 96-well plate that had been pre-plated using an automated liquid handler (Opentrons, Queens, NY, USA) under sterile filtered air flow. One hundred µL of CGM alone (negative control), Phytohemagglutinin-A/L (PHA-L) in CGM at 5 µg/mL (positive control), MARV peptides (Appendix A Table A4) at 10–20 g/mL, tetanus toxoid (positive control) (List laboratories, Campbell, CA, USA) at 2 g/mL, or tetanus peptide (positive control) (TT830–844) at 10 g/mL were added. Peptides used for macaque microsphere immunization were added to the wells at a concentration of 10 g/mL. All samples were assayed in duplicate. …

• Product #191 – Tetanus Toxoid from Clostridium tetani

Differentiation of SVF into adipocytes

When 100% confluency was reached, SVF cells were induced with the adipogenic cocktail containing 1 μM dexamethasone (Sigma-Aldrich, D4902), 10 μg ml−1 insulin (Sigma, I9278) and 0.5 mM isobutylmethylxanthine (Sigma, I7018) supplemented in DMEM/F12 for 2 days, followed by incubation in the adipocyte culture medium (DMEM supplied with 10% FBS) containing 10 μg ml−1 insulin for another 2 days. Then the cells were maintained in DMEM supplied with 10% FBS for 2 days. For treatment with inhibitors, cells were fasted for 12 h and pretreated with gallein (10 μM; MedChemExpress, HY-D0254), Y-27632 (10 μM; Solarbio, IY0050), SB203580 (10 μM; MedChemExpress, 152121-47-6), PTX (100 ng ml−1; List Biological Laboratories, 180) or H89 (20 μM; MedChemExpress, HY-15979A) for 4, 1, 1, 12 or 1 h, respectively, before treatment with 20 mM sodium L-lactate.

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