Bacterial Toxin Research Citations

EAE Induction and Clinical Scoring

EAE was induced after a minimum of 30 days post TAM administration by subcutaneous immunization at two sites of the chest with an emulsion that contained 100 μg MOG p35-55 (Genemed Synthesis) and Complete Freund’s Adjuvant containing 200 μg of heat-killed Mycobacterium tuberculosis (H37RA, Difco Laboratories). Mice were also injected with Bordetella pertussis toxin (PTX) (List Biologicals, dose used was lot-dependent, but varied between 100 and 250 ng) on days 0 and 2 following 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)

Stereotaxic Surgery

… For conventional retrograde tracing, rats were given unilateral microinjections of 100 nl of 2% Fluoro-Gold (FG, Fluorochrome, Denver, CO, USA) into unilateral NAc core (AP +1.8, ML +2.6, DV –7.3 from bregma, 6°) and 100 nl of 0.2% cholera toxin subunit b (CTB, List Biological Laboratories, Campbell, CA, USA) into RMTg (AP –7.2, ML +1.9, DV –7.6 from dura, 10°). FG was used only in NAc because it tended to cause toxicity in RMTg, evident by circling of animals after unilateral injection. Rats were sacrificed at least 1 week after injections. …

Tissue Processing

… For FG and CTB tracing, free-floating sections were incubated overnight in goat anti-CTB (1:50 K, List Biological Labs Cat# 703, RRID: AB_10013220) in PBS with 0.25% Triton X-100 …

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Mice were immunized subcutaneously at four injection sites with a total of 200 µl of emulsified incomplete Freund’s adjuvant supplemented with … and received intraperitoneal injections of 200 ng pertussis toxin (PTX; List Biological Laboratories) at the time of immunization and 48 h 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)

Flow cytometry of T cells

… For ICS, we stimulated cells with overlapping peptide pools (15 mers overlapping by 11 amino acids) targeting the S1 or S2 domains of spike glycoprotein, N, or E proteins (JPT Peptide Technologies). The S1 pool spans the N-terminal amino acid residues (1–643 amino acids, 158 peptides) of spike glycoprotein, while the S2 pool spans the C-terminal amino acid residues (633–1273 amino acids, 157 peptides). Each peptide pool was reconstituted with 40 or 50 μL of pure DMSO (Sigma-Aldrich) and then diluted with PBS for a final concentration of 100 μg/mL in 16% DMSO/84% PBS or 20% DMSO/80% PBS. PBMC were plated at a density of up to 1 × 106 cells/well in a 96-well U-bottom plate and stimulated with 1 μg/mL of each peptide in the pool, or 0.25 μg/mL Staphylococcal Enterotoxin Type B (SEB) (List Biological Laboratories Inc.) …

Mouse tracheal epithelial cell harvest and growth conditions

Mice of each genotype (WT, SP-A-/-, SP-A2 223Q, and SP-A2 223K) were euthanized with Urethane and tracheas were removed and placed in Ham’s F-12 media on ice. … The media used to culture the mouse tracheal epithelial cells (MTECs) consisted of Ham’s F-12 media supplemented with 250 ng/ml amphotericin B solution (Hyclone), 20 ng/ml cholera toxin (LIST biological), 104 μg/ml bovine pituitary extract (Lonza), 5 μg/ml insulin (Sigma), 5 μg/ml apo-transferrin (Sigma), 0.1uM dexamethasone (Sigma), 5ng/ml mouse epidermal growth factor (Sigma), 0.01 μM retinol (Sigma), 20 U/ml nystatin (Sigma), 50 μg/ml gentamicin. During the growth and expansion phase of culture, this media was further supplemented with 10% FBS. …

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

… emulsified with Freund’s Complete Adjuvant (CFA) containing 5 mg/mL heat-killed Mycobacterium tuberculosis H37RA (Difco Laboratories, Detroit, MI, USA) on day 0. In addition, 200 ng of Pertussis toxin (List Biological…

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. Reagents

… Ultrapure Lipopolysaccharide (LPS) was purified from Salmonella minnesota R595 (List Biological …

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

… A total of 50 µl emulsion was deposited on either side of the tail base. Pertussis toxin (300 ng per 200 µl; 180, List Biological Laboratories) was intraperitoneally injected on days 0 and 2 after MOG immunization …

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

2.5. Multiplex immunoassay

IgG concentrations specific to vaccine antigens PT, PRN, FHA, TT and DT were measured using an in-house multiplex bead-based immunoassay (MIA). IgG specific to FIM 2/3, which is not included in Boostrix, was also measured as a proxy for natural infection. PT, PRN and FHA were kindly provided by GlaxoSmithKline (Belgium). TT was purchased from Sigma-Aldrich. DT and FIM 2/3 were sourced from List Biological Laboratories (USA). A standard curve was generated using a 10-step 3-fold serial dilution of an in-house reference serum. Blanks and two quality control (QC) samples were included on every plate to calculate % critical variance across all assays, which fell between 5.1 and 14.6%. Assay specificity was determined using inhibition and interference assays. No cross reactivity was detected (Supplementary Figs. 1 and 2). …

• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #186 – Fimbriae 2/3 from B. pertussis

Antigens and biotinylation

… pertussix toxin (List Biological Laboratories), …

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)