Citations

Experimental human endotoxemia

All endotoxemia-related procedures were performed according to our standard continuous LPS-infusion protocol (12). In short, subjects were admitted to the research unit situated on the Medium Care department of the Radboud university medical center for approximately ten hours. Subjects had to refrain from consumption of alcohol and caffeine (24 h), and food and drinks (12 h) prior to LPS administration. Upon admission on the first LPS challenge day, two antebrachial venous cannulas were placed to allow administration of fluids, study medication and LPS. A radial artery catheter (BD Infusion Therapy Systems, Sandy, Utah, USA) was placed under ultrasound guidance to allow serial blood sampling and continuous hemodynamic monitoring. To reduce the risk of vasovagal responses (14), hydration fluids (2.5% glucose/0.45% sodium chloride) were administered as an initial prehydration bolus of 1.5L in the 45 min prior to LPS administration, and at a rate of 150 mL/hr for the remainder of the experiment. After prehydration, a bodyweight adjusted bolus dose of 1 ng/kg LPS (E. coli type O:113, lot no. 94332B1; List Biological Laboratories, Campbell, USA) was administered intravenously, directly followed by continuous infusion of LPS at a rate of 0.5 ng/kg/hr for three hours. Blood samples were serially obtained to construct time-concentration curves of circulating cytokines. Heart rate and intra-arterial blood pressure were measured using a 4-lead electrocardiogram (M50 Monitor, Philips, Eindhoven, the Netherlands) and a radial artery pressure transducer (Edwards Lifesciences, Irvine, California, USA). Hemodynamic parameters were recorded every 5 s using in-house developed software. Every 30 min, body temperature was measured using a tympanic thermometer (FirstTemp Genius 2, Covidien, Dublin, Ireland). Flu-like symptoms (headache, nausea, cold shivers, myalgia and back pain) were scored every 30 min using a numeric six-point Likert scale (0 = no symptoms, 5 = worst ever experienced). A composite score was then calculated as the sum of all previous elements, resulting in a total symptom score ranging from 0 to 25. Furthermore, general malaise was scored on an eleven-point scale (0 = no symptoms, 10 = worst ever experienced).

• Product #9433 – GMP LPS Lipopolysaccharide List™ Hpt™ From Escherichia Coli Type O113

The induction of the active immunization NMDAR mouse model

C57BL/6 mice (10 weeks old, female) were immunized with GluN1356–385 extracellular peptides, which were emulsified in complete Freund’s adjuvant (CFA) and supplemented with mycobacterium tuberculosis H37Ra (4 mg/ml), and the final peptide concentration was 1 mg/ml. Mice were injected subcutaneously on the back with 200 μg of peptide in the emulsion mixture and received two enhanced injections with peptide emulsion at 4 and 8 weeks after the first immunization. Mice in the control group received an emulsion mixture of CFA and an equal volume of phosphate-buffered saline (PBS). All mice were intraperitoneally injected with 200 ng pertussis toxin (List Biological Laboratories) on the day of the last immunization and 48 h 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)

… All mice then received an intraperitoneal injection of Pertussis toxin (List Biological Laboratories, #180) at a concentration of 2 ng/µL in 200 µL of PBS at the day of EAE induction and 2 days later. …

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

We used the following antibodies: … Goat anti-cholera toxin subunit B (List Labs – Cat#: 703); …

… In our experiments, this antibody strongly amplifies the signal that is produced in mice where Cre drives the production of EGFP in specific neuronal populations (GlyT2Cre;Ai140; Ai140 from Jackson mouse line 030220). Goat anti-cholera toxin subunit B (List Labs – Cat#: 703; RRID AB_10013220) is a goat polyclonal antibody against the Cholera Toxin B subunit.

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

… After 30 days, these animals were administered diphtheria toxin (DT, 50 ng/g BW, List Biological Lab, California) for three continuous days to deplete microglia (Parkhurst et al., 2013). …

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

HIV-1-specific T cell responses and assessment of T cell function

Thawed PBMCs were rested overnight in RPMI-1640 medium with 10% fetal bovine serum a. The following day we stimulated 1.5 x
106 PBMCs for 6 h in the presence of 2 mg/mL Gag potential T cell epitopes (PTE) peptides pool (AIDS Reagent Program, Division of AIDS, National Institute of Allergy and Infectious Diseases [NIAID], National Institutes of Health [NIH]. 1.5 x 106 PBMCs were simultaneously stimulated with staphylococcal enterotoxins B (SEB; List Biological Laboratories) at 1 mg/mL and another aliquot in the presence of media alone as the negative control. …

In vivo treatments with chemicals and antibodies

For DT-induced depletion of IM, IMDTR mice were injected i.p. with a single dose of 50 ng DT (List Biological Labs, 150), unless otherwise stated. …

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

CD115+ M-MDSC depletion

MMDTR mice were implanted with 2.5 × 105 EL4 cells. Each mouse received one intraperitoneal injection (10 ng/g of body weight) of diphtheria toxin (DT) (List Biological Laboratories, Campbell, CA, USA) on days 7, 9, and 11 after tumor implantation (Supplementary Fig. 7b). Time intervals (every 48 h) of DT injection were optimized by flow cytometry blood analysis (Supplementary Fig. 7a). After sacrifice on day 14 after tumor implantation, single cells were prepared for flow cytometry analysis. CD11b+Ly6G+Ly6Clo cells from naive or TB mice were sorted for OLFM4 immunohistochemistry (IHC) and qRT-PCR analysis. C57BL/6 mice were used as a negative control to exclude effects related to DT.

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

2.2.1. Antigens and Couplings

Tetanus toxoid was purchased from Massachusetts Biological Laboratories (Boston, MA, USA) and diphtheria toxoid was purchased from List Biological Laboratories (Campbell, CA, USA). Antigens were coupled to MagPlex microspheres (Luminex Corp, Austin, TX, USA), as described previously [5,23], using antigen concentrations of 12.5 µg/12.5 × 106 microspheres for tetanus toxoid and 60 µg/12.5 × 106 microspheres for diphtheria toxoid.

• Product #151 – Diphtheria Toxoid, from Corynebacterium diphtheriae

… Anthrax Protective Antigen List Biological Labs Cat# 171E Anthrax Lethal Factor List Biological Labs Cat# 172C …

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