Citations

2.1.1. Experimental Materials

The following materials were used: anthrax protective antigen (PA), from List Labs in California, USA, stored at 4 °C; anti-protective antigen (Anti-PA) monoclonal antibody, from List Labs in California, USA, stored at 4 °C; …

… Pertussis toxin (300 ng per 200 µL; 180, List Biological Laboratories) was intraperitoneally injected on days 0 and 2 after MOG immunization. Daily monitoring of EAE mice …

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

Induction of EAU model

On day 0, we injected mice subcutaneously with a 1:1 mixture of 200 μg retinal antigen interphotoreceptor retinoid-binding protein 1–20 (IRBP1–20) (amino acid sequence: GPTHLFQPSLVLDMAKVLLD; GL Biochem, Shanghai, China) and complete Freund’s adjuvant (Difco, Detroit, MI, USA) containing 2.5 mg/mL of Mycobacterium tuberculosis strain H37Ra (BD Difco, San Jose, CA, USA). After immunization, 0.25 ug pertussis toxin (PTX) (List Biological Laboratories, Campbell, California, USA) dissolved in phosphate-buffered saline (PBS) was injected intraperitoneally 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)

… Primary antisera contained antibodies raised against PHAL (species: rabbit; dilution: 1:4,000; Vector Labs Cat # AS-2224); RRID: AB_2313686) or CTB (species: goat; dilution: 1:10,000; List Biological Cat # 703; RRID: AB_10013220) …

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

2.5. Experimental human endotoxemia

On days 0 and 7, experimental endotoxemia was performed according to our standardized protocol8. Briefly, subjects were admitted to the research unit of the Radboud university medical center for eight hours, where a radial artery catheter (BD Infusion Therapy Systems) and antebrachial venous cannula were placed to allow serial blood sampling, hemodynamic monitoring and administration of fluids and LPS, respectively. In the 45 minutes prior to LPS administration, hydration fluids (2.5% glucose/0.45% sodium chloride) were administered as a 1.5L prehydration bolus to reduce the risk of vasovagal collapse13, and thereafter at a rate of 150 mL/h for the remainder of the experiment. Directly after prehydration, a bodyweight-adjusted bolus dose of 1 ng/kg LPS (E. Coli-derived, Type O113, lot no. 94332B1; List Biological Laboratories) was administered.

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

2.2. EAE induction and assessment

EAE was triggered using a four-point paraspinal subcutaneous injection of an emulsion containing complete Freund’s adjuvant (CFA) (0.05 mL, Sigma, St Louis, USA), myelin oligodendrocyte glycoprotein (MOG) 35–55 peptide (250 μg, Peptide Biotechnology, Nanjing, China), 0.05 mL of sterilized saline and Mycobacterium tuberculosis H37Ra (4 mg/mL, Difco Laboratories, Detroit, USA); subsequently, pertussis toxin (500 ng, List Biological, CA, USA) was given through intraperitoneal injection, and again the same amount was injected after 48 h. …

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)

… Recombinant, nicked PA63 from B. anthracis was obtained from List Biological Laboratories Inc., Campbell, CA. …

• Product #174 – Anthrax Protective Antigen, Activated (PA 63) from B. anthracis

Cells and reagents

… 1 mM intermittently vortexed and incubated in warm media [37°C] until soluble), or LPS (433, List Biologicals, suspended in media) as detailed in representative figures. …

• Product #433 – LIST™ HPT™ from Escherichia coli O113

2.14. Cell depletion in mice

Diphtheria toxin receptor (DTR)-transgenic mice were intraperitoneally administered 40 μg/kg of diphtheria toxin (DT) (List Labs, Campbell, CA, USA) to deplete plasmacytoid dendritic cells (pDCs) in BDCA2-DTR mice or CD169+ cells in CD169-DTR mice. To deplete lung macrophages, mice were intranasally (100 μl/mouse) and intravenously (200 μl/mouse) treated with clodronate liposomes (Liposoma BV, Amsterdam, Netherlands).

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

Immunization of mice with the H2Kb-restricted SIINFEKL peptide together with the CASAC

C57BL/6 mice were immunized intraperitoneally (i.p.) with CASAC (43) to the H2Kb-restricted SIINFEKL peptide containing the following reagents and their amount per mouse in double-distilled water: antigen peptide, SIINFEKL (100 μg; AS-60193-1 Eurogentec); stimulating antibody, anti-CD40 (clone 3/23, 25 μg; MCA1143 Bio-Rad); T helper 1 cell cytokine, IFN-γ (100 ng; Peprotech); Toll-like receptor 4 (TLR4) agonist emulsion, monophosphoryl lipid A + trehalose 6,6-dimycolate emulsion (Sigma Adjuvant System, used as per manufacturer’s instructions; Sigma-Aldrich); and TLR9 agonist, CpG 1826 (25 μg; Invivogen). Final vaccine volumes were 200 μl. Vaccines are warmed to 37°C and shaken vigorously before injection. All mice are injected i.p. with PT (400 ng; List Biological Laboratories) 24 and 72 hours after 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)