Citations

Induction of EAE in mice

EAE mice were induced with MOG35-55 (200 µg), and the mice were intraperitoneally injected with pertussis toxin (500 ng, List Biological Laboratories, Inc.) at 0 and 48 h following immunization. At least two investigators weighed and evaluated the animals for clinical scores in a blinded manner. There were two animals which appeared to be in intolerable distress and self-mutilated limbs; these were sacrificed using pentobarbital sodium (150 mg/kg).

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)

… One hundred nanograms of pertussis toxin (List Biological Labs, Campbell, CA, USA) in 0.1 mL saline was also injected intraperitoneally on day 0, and 48 hours 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)

Antibody measurements:

For PT-specific IgG ELISA, plates were coated with 0.5µg/mL purified PT (List Biologicals Campbell, California) in PBS and the assay performed as previously described [133]. …

Author did not specify which List Labs Pertussis Toxin was utilized.  List Labs provides Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer and Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free).

… The effect of Ex-4 on cytokine levels in microglia cell cultures was also studied. Whole brains of young mice (day 2-3 after birth) were homogenized and microglia cells cultured and enriched. They were then stimulated with lipopolysaccharide (LPS) (10 ng/mL, List Biological Laboratories Inc., Cambell, CA) with or without Ex-4 (10 nmol/L). …

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

… During the final 8 weeks, mice were challenged with 10 ng of Staphylococcal enterotoxin B (SEB; Product# 122; List Biological Laboratories Inc., Campbell, CA, USA), diluted in 1 PBS, immediately after the instillation of 3% OVA. …

STAR★Methods
Key Resources Table

Cholera toxin (Azide-free) from Vibrio cholerae List Biological Laboratories Cat#100B

Oral Immunization and ELISA

Before immunization with CTX, mice were deprived of food for 2 h and then given 0.25 mL of a solution containing eight parts of Hanks balanced salt solution (HBSS) and two parts of 7.5% sodium bicarbonate by oral gavage to neutralize stomach acidity. After 30 min, mice were orally immunized with 30 μg CTX (List Biological Laboratories). Serum and fecal pellets were collected 14 days after immunization and were analyzed by ELISA as previously described (Ushijima et al., 2018). Briefly, 96-well polystyrene plates (Thermo 3855) were coated overnight at 4°C with CTX (0.5 μg/well; List Biological Laboratories) or OVA (0.5 μg/well; Sigma-Aldrich). After the wells were blocked with PBS containing 1% sodium casein (Wako) and 0.1% Tween-20 (Wako), serial dilutions of sera were added to the well. …

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

Recombinant antigens and coupling to microsphere beads.

… Tetanus toxoid (Massachusetts Biological Laboratories, Boston, MA), diphtheria toxoid from Corynebacterium diphtheriae (List Biological Laboratories, Campbell, CA), and recombinant measles nucleoprotein (MV-N, Meridian Life Sciences, Memphis, TN)31 were purchased from commercial sources. Tetanus toxoid was coupled to SeroMap beads as previously described.32 Diphtheria toxoid was coupled in buffer containing 50 mM 2-(N-morpholinoethanesulfonic acid (MES) at pH 5.0 with 0.85% NaCl at a concentration of 60 µg of protein per 1.25 × 107 beads in 1 mL final volume. …

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

2.11 EAE induction

EAE was induced by subcutaneous injection of 150 μg MOG peptide (amino acids 35‐55), in 0.1 mL of complete Freund’s adjuvant (CFA) emulsion containing 400 μg Mycobacterium tuberculosis (Difco Laboratories, Detroit, MI, USA), into wild‐type or Iqgap1−/− B6 mice on day 0. The mice received intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories, Campbell, CA, USA) on days 0 and 2. …

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)

2.9. Experimental Autoimmune Encephalomyelitis (EAE) Induction in a Murine Model

In order to mimic the pathology of human MS, EAE was induced using a murine model as previously described [27,28]. Three-month old C57BL/6J mice (male and female) were immunized with myelin oligodendrocyte glycoprotein (MOG) peptide 35–55 to induce EAE (IACUC #19014). In brief, 300 μg of rodent MOG peptide (amino acids 35–55, New England Peptides, Gardner, MA, USA) in Complete Freund’s Adjuvant (CFA) containing 5 mg/mL killed Mycobacterium tuberculosis (Difco, Thermo Fischer Scientific) was administered into the subcutaneous flank of mice at day 0. At day 0, each mouse received two subcutaneous injections of the MOG solution as well as a 100 µl dose of 2 ng/µl pertussis toxins and virulence factors (List Biological Laboratories Inc., Campbell, CA, USA) diluted in sterile PBS (Thermo Fisher Scientific). Pertussis toxins and virulence factors were administered again on day 2. Pertussis toxins allow an increase in the blood–brain barrier permeability in order to facilitate the incursion of the different treatments into the CNS. …

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)

Additional Pertussis products can be found at https://listlabs.com/products/bacterial-toxins/?sub=pertussis-toxin-and-virulence-factors

… tetanus toxoid (TT) (List Biological Laboratories, Campbell, CA). Peptides were plated at 10 g/ml, TT at 1 g/ml, ECD at 25 ng/ml and …

• Product #191 – Tetanus Toxoid from Clostridium tetani