Bacterial Toxin Research Citations

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

Reagents. LPS from E. coli serotype 0111:B4 was purchased from List Biological Laboratories (Campbell, CA). …

Chromatin Preparation and ChIP Assay.

Macrophages were stimulated in the presence or absence of LPS (10 ng/mL) in RPMI containing 10% FBS for up to 8 hours. …

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

Establishment of the EAE model

According to a previously established method30, WT C57BL/6 mice were subcutaneously immunized with 200 μg of murine myelin oligodendrocyte glycoprotein (MOG)35–55 peptide (Catalog Number: MOG-001, Chinese Peptide Co., Ltd., Hangzhou, China) in complete Freund’s adjuvant (Catalog Number: 7001, Chondrex Inc., Woodinville, WA, USA). Each mouse was intraperitoneally injected with 400 ng of pertussis toxin (Catalog Number: 181, List Biological Laboratories, Campbell, CA, USA) twice at 0 h and 48 h postimmunization. …

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

Toxin challenge

In vivo challenge experiments with CT were performed as described previously (Tokuhara et al., 2010). Briefly, vaccinated mice (10 mice per group) were orally administered 20 μg CT (List Biological Laboratories, Campbell, CA, USA); 12 h later, mice were euthanized and their small intestine and colon were removed for clinical evaluation of diarrhea and collection of intestinal contents. Samples were centrifuged and intestinal water content was measured.

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

Retrograde viral tracing from the trachea and the lungs
… In the subsequent confirmatory experiments, the Cre-dependent viral tracer, AAVrg-CAG-FLEX-tdTomato-WPRE10GH, was injected into the trachea of heterozygous CGRP-Cre mice (n = 2). Again, animals were anaesthetized with isoflurane (4% for induction and 1.8–2.5% for maintenance), a midline incision was made in the neck and the muscles overlying the trachea were retracted to expose the trachea. Next using a Hamilton syringe, 1 μl of AAVrg-CAG-FLEX-tdTomato-WPRE10GH together with 2μl of saline was injected into the lumen of the trachea. After 4 weeks, the animals were euthanized and their brains, trachea and vagal ganglia were collected for immunohistochemistry. Alternatively, cholera toxin B (CTB, catalogue number #104, List Biological Labs, 0.3% w/v, 5 μl per injection), a retrograde tracer, was injected into the trachea of Phox2b-Cre;loxP-ChR2 mice (n = 6) following the experimental procedures described above. After one week, the animals were euthanized and their brains and vagal ganglia were collected for immunohistochemistry.

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

Immunizations, oral tolerance induction, and FTY720 treatment

To induce local IgA immunity mice were fed 10 mg of ovalbumin (OVA) protein grade V (Merck KGaA, Darmstadt, Germany) dissolved in 0.5 mL of phosphate buffer saline (PBS) containing 3% (w/v) sodium bicarbonate with or without the addition of 10 µg cholera toxin (CT) adjuvant (List Biological Laboratories, Campbell, CA, USA) to each dose. The mice were given three oral administrations, 10 days apart, using intragastric intubation. To induce oral tolerance, mice were fed three doses of 20 mg OVA dissolved in 0.5 mL of PBS containing 3% (w/v) sodium bicarbonate 7 days apart. …

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

Brain Imaging

Mice were anaesthetized with isoflurane and rectal temperatures recorded before being inoculated i.t. with either EPS+, EPS- or 50 μg ultra-pure lipopolysaccharide from Escherichia coli O111:B4 (ListLabs; #421) in a final volume of 50 μl PBS. At 4 h post infection, the mice were anaesthetized under isoflurane and rectal temperatures recorded before intravenous perfusion with cold PBS. The mice were then perfused with cold 4% formalin. The brains were then harvested and stored in 4% formalin for 24 h at 4 °C. After 24 h, the brains were then washed with PBS 3 times (10 minutes each) while swirling. Brains were then stored in 20% sucrose and kept at 4 °C.

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

Phagocytosis Marker Assay

BMDMs were stimulated with LPS at 100 ng/mL (ULTRA PURE LPS from Escherichia coli O111:B4, #421, List Labs) or saline vehicle for 4 hours. …

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

2.2 DT or saline administration

Adult mice (6–16 weeks of age) received two intramuscular (IM) injections of DT (List Biological Laboratories, Campbell, CA) at either 25 ng/g (low-dose) or 50 ng/g (high-dose), spaced 2 days apart. Some control mice were administered IM saline.

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