Bacterial Toxin Research Citations

AD models

For the mouse AD model, the mice were epicutaneously sensitized with HDMs and SEB as described previously [33] or treated with MC903 [46, 47].

In brief, a solution of 10 μg of HDM extract (Stallergen Greer) and 500 ng of SEB (List Biological Laboratories) in PBS was applied to a 1 cm2 gauze pad placed on the shaved skin and occluded with a Tegaderm Transparent Dressing (3 M Healthcare). Three days later, the gauze pads were replaced. …

Immunizations and EAE induction

Mice were immunized subcutaneously in the flanks with 100 µg of PLP178-191 or OVA323-339 emulsified in Complete Freund’s Adjuvant (CFA) supplemented with 4 mg/ml Mycobacterium tuberculosis (Becton Dickinson, Franklin Lakes, NJ). The day of immunization was considered day 0. For EAE induction, mice immunized with PLP178-191 received two doses of 250 ng of Pertussis Toxin (PTX) in glycerol (List Labs, California) on days 0 and 2 intraperitoneally. …

• Product #179A – Pertussis Toxin from B. pertussis (in Glycerol)

Tissue processing: … The following primary antibodies and dilutions were used: mouse anti-laminin (1:50, Sigma-Aldrich, Burlington, MA, USA, #CC095-M), goat anti-CTB (1:200, List Biological Laboratories, Campbell, CA, USA, #703), …

• Product #703 – Anti-Cholera Toxin B Subunit (Goat)

… Anti-PA IgG was measured in rabbit sera via electrochemiluminescence similar to the PA-ECL screening assay. Biotinylated recombinant PA83 (List Biological Laboratories, Inc.) was bound to streptavidin-coated plates (MSD) and used as the capture antigen. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

Immunization, Nb infection and in vivo treatment

All immunization procedures were conducted in the rear footpad with 20 µL injection volume per footpad. Mice were immunized as indicated in each Figure with 5 µg low-endotoxin OVA (Worthington Biochemical Corporation) mixed with either 50 µg papain (P4762, Sigma), 10 µL FCA (F5881, Sigma), 10 µL alum (77161, Thermo Scientific), or 10 µg CpG2216 (Invivogen) in phosphate-buffered saline (PBS). For Nb infection, mice were injected subcutaneously in the right ear with 600 third-stage (L3) larvae (gift from William Gause, Rutgers New Jersey Medical School) in 0.2 mL of PBS. For DC depletion, mice were injected i.p. with 500 ng DT in PBS (List Biological Laboratories) at indicated time-points. …

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

STAR★Methods
Key resources table

PTX (100 ng/mL) List Biological Laboratories Cat# 180

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

STAR★Methods
Key resources table

Pertussis toxin List Biological Cat# 181

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

Preparation of DT-conjugated latex beads and ICV injection

Two microliters of 1 μm carboxylated latex beads (Sigma-Aldrich; L4655) were diluted to 40 μl in ACSF (artificial cerebrospinal fluid)(Coolaber, SL6630-500ml), followed by co-incubation with 160 μl ACSF containing 100 μg of DT (List labs; 150) in a light-protected tube with continuous vortex overnight at room temperature. After that, beads were washed with cold PBS for three times to remove unbound free DT. Finally, the DT-conjugated beads were resuspended in 40 μl PBS. BSA-conjugated beads were used as controls. Beads were injected ICV at a volume of 5 μl per mouse at 1 μl min−1.

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

Colony-forming assay

Colony-forming assay (CFA) was performed following the protocol previously reported11,50. Briefly, trypsinized limbal epithelial cells were seeded on the 3T3-J2 feeder cell layer treated with mitomycin C (MMC) (MilliporeSigma) at 500 cells per well on 6-well plates and were cultured for 10 days in keratinocyte culture medium (KCM) supplemented with 10 ng/ml KGF and 10 µM Y-27632. KCM consists of DMEM without glutamine and Ham’s F-12 Nutrient Mix (Thermo Fisher Scientific) combined in a ratio of 3:1, supplemented with 10% FBS, 0.4µg/ml hydrocortisone hydrogen succinate (MilliporeSigma), 2nM 3,3’,5-triiodo-l-thyronine sodium salt (MilliporeSigma), 1 nM cholera toxin (List Biological Laboratories, Campbell, CA), 2.25 µg/ml bovine transferrin HOLO form (Thermo Fisher Scientific), 2mM L-glutamine (Thermo Fisher Scientific), 0.5% (vol/vol) insulin transferrin selenium solution (Thermo Fisher Scientific), and 1% (vol/vol) penicillin-streptomycin solution GE Healthcare Life Sciences). …

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

Animal surgeries

… For central lesions, wild-type and Spastin knockout mice underwent spinal cord dorsal hemisection at the T8 level using a micro-feather ophthalmic scalpel (#72045-45, Delta Microscopes). For conditioning lesions, a bilateral complete sciatic nerve transection was performed first, followed a week later by spinal cord dorsal hemisection. Regeneration of central axons was evaluated after 6 weeks post-spinal cord injury. During this period, animals received analgesia, and bladder voiding was supported twice daily via gentle abdominal compression. To trace regenerating dorsal column axons, 2 μl of 1% cholera toxin-B (List Biologicals, 103B) was injected into the left sciatic nerve with a 10 μl Hamilton syringe 4 days before euthanasia. …

Analysis of regeneration of dorsal column tract axons

Serial sagittal sections of the spinal cord (50 µm) from non-injured mice, spinal cord-injured mice, and mice where the spinal cord injury was preceded by a sciatic nerve injury (conditioning group) were collected for immunofluorescence against cholera toxin-B. The sections were blocked and incubated overnight with goat anti-cholera toxin-B primary antibody (1:30.000, #703, List Biologicals, RRID:AB_10013220), followed by …

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