Bacterial Toxin Research Citations

EAE induction

All animal studies are approved by the Johns Hopkins University Animal Care and Use Committee. Animals received standard husbandry. C57Bl/6 mice (female, 6–10 weeks, Jackson Laboratories, n = 5–8) were injected s.c. with 200 μl of emulsion containing incomplete Freund’s adjuvant (Sigma F5506) Mycobacterium tuberculosis H37Ra (Difco BD 231141, 5 mg/ml) and myelin oligodendrocyte glycopeptide (MOG35-55, Johns Hopkins Synthesis and Sequencing Facility, 0.5 mg/ml). Mice were injected i.p. with 100 μl of pertussis toxin (List Laboratories #180, 300 ng) on the day of induction and 2 days later. …

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

… To deplete cDC1s in XCR1DTR mice, diphtheria toxin (DT; List Biological Laboratories) at 500 ng/mouse was injected intraperitoneally every three days beginning on day …

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

… For imaging, cells were seeded in 96 well μClear® plates (Greiner Bio-One) or NuncTM MicroWellTM optical bottom 96-well plates (Thermo Fisher Scientific) and allowed to establish a confluent monolayer 16–24 h prior to evaluation by high content or scanning confocal microscopy, respectively. For immunofluorescence analysis, cells were seeded on a 12-mm microscope cover glass (VWR) in a 24-well plate overnight. Where indicated, BMMØs were cultured in the presence of either 10 ng/mL recombinant murine IL-4 or 100 U/mL recombinant murine IFN-γ (PeproTech) for 24 and 18 h respectively, prior to phagosomal assessment. Where indicated, cells were pulsed overnight with 2 mg/mL Bella Fluor (BF) 350SE-labelled (Setareh Biotech) dextran (70 kDa) with the dextran being chased into lysosomes the following day for 2 h. Where indicated, 2.5 mM of LLOMe was added to cells 30 min after phagocytosis of experimental particles and incubated for 2 h prior to imaging. Where indicated, 10 nM of myricetin (Alfa Aesar), 10 nM of kaempherol (Selleckchem) or 100 nM of EACC (Life Chemicals) were added to cells at the same time as experimental particles. Where indicated 0.5 μM Torin 1 (Tocris Bioscience), 0.5 μM rapamycin (Sigma Aldrich), 1× MEM amino acid solution (Sigma Aldrich), 10 ng/mL ultrapure lipopolysaccharide from S. Minnesota R595 (LPS; List Biologicals) were added to assay medium 30 min after the addition of reporter particles or 50 nM Lysotracker® Red (Invitrogen) 2 h after the addition of experimental particles and remained in the assay medium for the duration of phagosomal assessment.

• Product #434 – ULTRA PURE LPS from Salmonella minnesota R595 (Re)

… LPS from Salmonella Typhimurium was purchased from List Biological Laboratories …

• Product #225 – LPS from Salmonella typhimurium

… Pertussis toxin (300 ng per mouse; List Labs, Campbell, CA, USA) was injected intraperitoneally on the same day and 2 days later. Mice were scored daily for clinical symptoms from 0 to 5 with 0.5 gradations: 0, no detectable signs; 1, limp tail; 2, limp tail …

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)

… For anti-filamentous hemagglutinin (FHA) or anti-pertactin (PRN) ELISAs, CoStar 96 well high-binding
plates (Corning, Corning, NY) were coated with either 2 µg/ml FHA (BP-FHA-100, The Native Antigen
Company/Cedarlane, Burlington, NC) or 1 µg/ml PRN (B. pertussis Pertactin, Strain: B. pertussis strain
165, The List Labs, St Campbell, CA) in carbonate buffer pH 9.6, incubated overnight at 4°C, washed 3x
with wash buffer (KPL 10X Phosphate Buffered Saline with 0.05% Tween 20 (Fisher Scientific)) and
blocked with Superblock (ScyTek) for 1 h at room temperature (RT). …

• Product #187 – Pertactin from B. pertussis (69 kDa Protein)

2.2. Antigens Vibrio cholerae toxin (CT holotoxin; lot 10154B5A; Mr 84,000) was from List Biological Labs (Campbell CA, USA) …

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

Cell culture

… Where indicated, cells were also treated with nigericin (5 μM, Invivogen, 10–45 min), CTB (20 μg ml−1, List Biological Laboratories, 6 h), silica (200 μg ml−1, Invivogen, 6 h), TcdB (10 μg ml−1, R&D Systems, 8 h) and ATP (5 mM, Millipore-Sigma, 25 min); cultured in K+-free medium (0.8 mM MgCl2, 1.5 mM CaCl2, 10 mM HEPES, 5 mM glucose and 140 mM NaCl, pH 7.2, 3 h)50; or transfected with flagellin (500 ng ml−1, Invivogen, 4 h), poly(dA:dT) (1 μg ml−1, Invivogen, 4 h) or FSL-1 (0.2 μg ml−1, Invivogen, 4 h) and ultrapure LPS (1 μg ml−1, 4 h) with Lipofectamine 2000 (Invitrogen) or as otherwise indicated.

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

… Stimuli: LPS O111:B4 (L3012, sigma), LPS O55:B5 (L2637, sigma), CTB (103B, List Biological Laboratories), …

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

… XCR1+ cDC1 cells were depleted by injecting Xcr1DTR mice with 500ng un-nicked DT (150, List Biologicals) intraperitoneally in 100µl PBS. DT was typically administered 2d before induction of MICB expression on tumour cells by doxycycline treatment and then every 48h until the study endpoint. …

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