Citations

Toll-like receptor (TLR) 4 and 5 activity assays

TLR4 and TLR5 activity assays were performed as described previously with minor modifications (14, 15). Briefly, HEK-Blue-hTLR4 cells and HEK-Blue-hTLR5 cells carrying a secreted embryonic alkaline phosphatase (SEAP) reporter construct were obtained from InvivoGen (CA). Cells were maintained in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS) and 0.5% penicillin/streptomycin with and without 0.2% Normocin (InvivoGen, CA) for TLR5 cells and TLR4 cells at 37°C with 5% CO2, respectively. Monolayers of 1 × 105 cells per well in a 96-well plate were incubated with media alone, PBS, and FlaA and FlaB proteins at different concentrations ranging from 10 pg/mL to 10 µg/mL for 24 h. E. coli lipopolysaccharide O111:B4 (List Biological Laboratories, Inc., CA) was added to HEK-Blue-hTLR4 cell as a positive control. …

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

Tdap-specific IgG and IgG subclass detection

All available leftover maternal and cord serum samples from both deliveries were tested at the Université Libre de Bruxelles (ULB). Total and subclass Tdap-specific IgG antibodies were quantified using an in-house bead-based multiplex immunoassay (28). In brief, serum samples were incubated with distinct fluorescent magnetic beads, individually coated with specific purified target antigens: PT (List Labs, #180), FHA (Sigma, #F5551), PRN (NAC, #NAT41666), diphtheria toxin (DT, List Labs, #151) and tetanus toxin (TT, MassBiologics, # LP1105P). Samples were measured at appropriate dilutions with a Multigam reference (CAF-DCF, Belgium), a WHO international standard NIBSC 06/140 and blanks included on each plate. Serum dilutions used to titrate IgG and IgG1 were 1:9000-1:18000-1:36000, 1:70-1:140-1:280 for IgG2 and IgG3, and 1:70-1:700-1:7000 for IgG4. After washing, the captured IgG was detected with R-phycoerythrin labelled antibodies specific for each isotype (IgG1-4). …

• Product #180 – Pertussis Toxin from B. pertussis, Lyophilized in Buffer
• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

Pertussis toxin List Biological Laboratories, Campbell, CA Cat# 181238A1

EAE induction

EAE was induced in female C57BL/6 mice (8–10 weeks of age).38 Mice were subcutaneously immunized with 200 mg of the MOG35–55
peptide in a 100-ml solution (containing 50% complete Freund’s adjuvant with 10 mg/ml of heat-killed M. tuberculosis H37Ra strain).
The MOG35–55 peptide was injected into areas that drained into the axillary and inguinal lymph nodes. Pertussis toxin (200 ng/mouse) was administered intraperitoneally on the day of immunization and again at 48 h later. Animals were examined for disease symptoms
daily. The evaluation of disease severityEAE induction. …

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

Induction of EAE

EAE was induced in male C57BL/6 J mice (Jackson Laboratories) at 8–12 weeks of age by subcutaneous immunization with an emulsion containing 200 μg of myelin oligodendrocyte glycoprotein (35–55) (MOG35–55) in saline and an equal volume of complete Freund’s adjuvant (CFA) containing 400 ng of mycobacterium tuberculosis H37RA (Difco Laboratories, Detroit, MI). All mice were administered 400 ng of pertussis toxin (List Biological, Campbell, CA) intraperitoneally (i.p.) at 0 and 48 h post-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)

4.2. Air Pouch Model

To evaluate the possible immunomodulatory effects of PHA 568487 in vivo, the air pouch model of acute inflammation was used [12,24]. At 8 weeks of age, male C57BL/6JRj mice (Janvier Labs, Le Genest-Saint-Isle, France) were injected with 3 mL of sterile air between the scapulae under
isoflurane anaesthesia. Three days later, the air pouch was re-filled with 3 mL of sterile air. On the sixth day, the mice received an intraperitoneal injection of either PBS (Thermo Fisher Scientific, Waltham, MA, USA) or PHA 568487 (Tocris Bioscience, Ellisville, MO, USA) at 12.4 μmol kg-1 (5 mg/kg) or 123.4 μmol kg-1 (50 mg/kg). 15 min later, all mice were injected with LPS (10 μg, List Biological Laboratories Inc., Campbell, USA) directly into the pouch. The mice were sacrificed six hours after the LPS-injection by an overdose of pentobarbital (Apoteket AB, Stockholm, Sweden). …

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

… Cells were stimulated with 100 ng/ml ultrapure Escherichia coli LPS (List Biological Laboratories, Campbell, CA) and then 5 mM ATP (SigmaAldrich, St. Louis, MO) for the last 30 min of culture, as indicated. “Mock” treatment was the addition of the equivalent volume of media (without parasites or LPS) to cells. At the indicated time point, monocytes were pelleted …

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

4.3. EAE induction

EAE was induced using 200 μg/dose of MOG(35–55) peptide (MDBiosciences, Redoxis exp.) or 100 μg MOG(35–55) (Genemed Synthesis, Inc., UCSF exp.) emulsified with complete Freunds Adjuvant (CFA) containing IFA (Difco) with M. Tuberculosis H37RA (300 μg or 800 μg H37RA/dose (BD Difco) in the Redoxis experiment (Exp. 1) or UCSF experiment (Exp. 2), respectively). Mice were anaesthetized with isoflurane and the emulsion was injected subcutaneously (volume 100 μl or 200 μl in the Redoxis experiment or UCSF experiment, respectively) in the flank. Pertussis toxin (Ptx, 200 ng/dose, MerckMillipore or List Biological) was administered intraperitoneally (i.p.) 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)

… Mycobacterium tuberculosis (strain H37Ra; Cat# 231141, Becton, Dickinson and Company, Franklin Lakes, USA), followed by intraperitoneal administration of 400ng of Pertussis toxin (List Biological Laboratories, Cat# 181, Campbell, USA) on day 0 and after 48?h (Bonfiglio et al., 2017). …

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

Reagents and Biological resources

… E. coli O111:B4 Lipopolysaccharide (LPS) was purchased from List Biological Laboratories (catalog #201). …

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

Retrograde Tracing

Injections of CTB (List Biological Labs, Campbell, CA) into the NAcc (A: 1.30, L: ±0.85, D: -3.80), CPVM (A: 0.26, L: ±1.25, D: -3.0), LHb (A: -1.4, L: ±0.75, D: -3.90), VTA (A: -3.3, L: ±0.4, D: -4.0), and DRN (A: -2.24, L: -1.1, D: -2.85 with 20° angle) were performed stereotaxically in WT and 15q dup mice: 50 μL of 0.5% CTB solution dissolved in saline was injected using a glass micropipette. The mice were anesthetized 5 days post-CTB injection, with an intraperitoneal injection of an anesthetic combination comprising 0.3 mg/kg medetomidine, 4.0 mg/kg midazolam, and 5.0 mg/kg butorphanol. Then, they underwent cardiac perfusion fixation with 4% PFA in 0.01 M PBS. The post-fixed brains were rinsed with saline and cryoprotected overnight with 30% sucrose in PBS at 4°C. Brain sections (50 μm) were cut using a sliding microtome (HM430; Thermo Fisher Scientific, Waltham, MA, USA). Immunostaining was performed as previously described.

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