Citations

Immunofluorescence Histochemical Staining of DMV and Analysis

Triple immunofluorescence staining of CTb, VGLUT1, and ChAT was conducted for CTb-injected rats. Sections containing DMV was processed for CTb, VGLUT1, and ChAT triple-immunofluorescence staining. Briefly, the sections were incubated at room temperature with a mixture of goat anti-CTb Ig (1:2,000; List Biological, Campbell, CA, USA), …

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

Reagents

… and pertussis toxin was obtained from List Biological Laboratories (Campbell, CA, 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)

Multiplex luminex immunoassays

Antigen-specific antibody responses were measured through a modified multiplex Luminex assay as previously reported (22, 45). Pertussis (PTM, PT, inactivated PT), tetanus, and diphtheria proteins, PRN, FHA, FIM2/3, adenylate cyclase toxin (ACT), lipooligosaccharide (LOS), tetanus toxoid (T), and diphtheria toxoid (d), were purchased from List Biological Laboratory (Campbell) and …

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)
• Product #184 – Pertussis Toxin Mutant
• Product #188L – Adenylate Cyclase Toxin, Recombinant from B. pertussis
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae
• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae
• Product #190 – Tetanus Toxin from Clostridium tetani
• Product #191 – Tetanus Toxoid from Clostridium tetani

… Azide-free cholera toxin (CT: #100B) from Vibrio cholerae was obtained from List Biological Laboratories (Campbell, CA, U.S.A.) and used as the adjuvant. …

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

STAR★Methods
Key Resources Table

Pertussis toxin List Biological Laboratories Cat# 180(LB)

EAE induction and monitoring

For adoptive transfer of effector T cells, 5×106 Th17 or Th1 1C6 T cells were injected i.v., in cold PBS into NOD.Scid recipient mice, which additionally received 200 ng of pertussis toxin (List Biological Labs) i.p. on d0 and d2 post-injection. FCG B6 x NOD F1 mice were actively immunized with 200 μg MOG[35-55] in 200 μL of incomplete Freund’s adjuvant supplemented with 500 μg M. tuberculosis extract, and additionally received 200 ng pertussis toxin (List Biological Labs) on d0 and d2. Th1 and Th17 cells generated from FCG B6 × 1C6 mice were adoptively transferred to NOD.Scid at 2 × 106 cells/recipient. …

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

STAR★Methods
KEY RESOURCES TABLE
REAGENT or RESOURCE SOURCE IDENTIFIER

Chemicals, peptides, and recombinant proteins

Lethal factor List Biologicals Cat#172B

Protective antigen List Biologicals Cat#171E

Activation of BMDMs and human macrophages

All cells were grown, maintained and activated at 37°C and 5% CO2 in a water-jacketed incubator. 1 × 106 WT or Casp1/11 DKO murine macrophages were plated per 12-well in DMEM, 10% (v/v) FCS (DMEM complete medium) one day prior to stimulation. All BMDMs were primed with 200 ng/mL LPS from Salmonella Typhimurium for 2 h or left untreated and subsequently washed three times with serum- and phenol red-free DMEM supplemented with 2 mM L-glutamine (DMEM minimal medium). For the time course experiment (Dataset 1), cells were preincubated with 0 or 100 μM glyburide (#NBP2-30141, Novus Biologicals) for 15-30min before 2 mM ATP (#7699, Sigma) or 10 μM nigericin (N1495, Invitrogen) were added for 15, 30, 45, and 60 min. Lethal factor and protective antigen were added at 1ug/ml. …

• Product #171E – Anthrax Protective Antigen (PA), Recombinant from B. anthracis‘
• Prodcut #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

ELISA

The three strains cultured in TY medium for 12, 24, 48, and 72 h were centrifuged to collect the supernatants containing TcdA and TcdB toxins; the supernatant was filtered with a 0.45 µm filter. Next, a 96-well plate was coated with the supernatant at 4 °C overnight and then washed with PBST (phosphate-buffered saline pH 7.4 + 0.05% Tween 20). The plate was blocked with 5% skim milk at 37 °C for 2 h. The polyclonal antibodies of TcdA and TcdB (chicken IgY; 1:1000 dilution; List Biological laboratories) were used for first hybridization; then, goat anti-chicken IgY secondary antibody, (HRP-conjugated; 1:5000 dilution; Invitrogen) was used. The TMB chromogenic reagent kit (Sangon, China) was used to determine the absorbance at 450 nm. Pure toxin A and toxin B (List Biological laboratories) were used as positive controls, and the standard curves were obtained using 20, 2, 0.2, 0.02, and 0.002 ng/ml of TcdA and TcdB [19].

• Product #152C – Toxin A from Clostridium difficile
• Product #155 – Toxin B from Clostridium difficile

Induction and assessment of EAE

C57BL/6 mice were immunized by subcutaneous injection with 150 μg of MOG35-55 peptide (MEVGWYRSPFSRVVHLYRNGK, Sigma-Aldrich) emulsified in a complete Freund’s adjuvant containing 5 mg/ml H37RA (Difco Laboratories). On day 0 and 2 after immunization, 200 ng pertussis toxin (List Biological Laboratories) was given intravenously. …

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)

EAE Induction and Treatment

EAE was induced in 8-week-old mice by subcutaneous immunization with 150 μg MOG35–55 peptide emulsified in complete Freund adjuvant (Difco Laboratories, Detroit,
MI) per mouse, followed by administration of 200 ng pertussis toxin (List biological Laboratories, Inc., Campbell, CA) on days 0 and 2 as described. …

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)

… In each of three locations, 100 µl volume was injected s.c. Pertussis toxin (200 ng in 100 µl PBS; List Biological Laboratories) was injected i.p. on the day of 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)