Bacterial Toxin Research Citations

Alcohol administration and intratracheal injection:

Mice received intraperitoneal (i.p.) injection with ethanol [4.0 g/kg; 20% v/v ethanol in sterile phosphate buffered saline (PBS)] or an equivalent volume of PBS. Thirty minutes after i.p. injection, animals were anesthetized with isoflurane for exposure and direct intratracheal (i.t.) administration of 10 g Escherichia coli LPS (List Biological Laboratories; Campbell, CA). n 5 per group for all in vivo experiments.

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/

Samples of LF used for comparison were purchased from List Biological Laboratories (Campbell, CA).

Results – Toxicities of LF proteins produced from avirulent B. anthracis strains:

… B. anthracis strains cured of the pXO1 and pXO2 virulence plasmids [18]. LF expressed from pSJ115 is here termed LF-HMA to denote the presence of the two residues His-Met (HM) added at its N-terminus due to the cloning manipulations. This expression system is licensed to List Biological Laboratories (Campbell, CA), and the LF sold by them is therefore also LF-HMA. …

• Product #172 – Anthrax Lethal Factor (LF), Recombinant from B. anthracis

Materials:

SNAPtide substrate #520 contains the FRET pair o-aminobenzoic acid/2,4 dinitrophenyl (o-Abz/Dnp) and SNAPtide substrate Mca/Dnp contains (7methoxy-coumarin-4-yl)acetyl/2,4 dinitrophenyl as its FRET pair. Botulinum Neurotoxin Type A (Product #130A) and the SNAPtidesubstrates are products of List Biological Laboratories.

• Product #520 –  SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #130A – Botulinum Neurotoxin Type A from Clostridium botulinum

Cells and reagents:

… Escherichia coli LPS 0111:B4 was purchased from List Biological Laboratories, …

… On day 4, the BM cells were treated with 0 or 50 mM EtOH for another 5 days. On day 9, the cell density was regulated to 1 10⁶/ml/well in 24-well plates. After culturing overnight, medium was changed and cells were incubated for additional 6 h with 0 or 100 ng/ml LPS (E. coli 0111:B4; List Biological Laboratories). …

TNF- mRNA half-life assay:

A total of 1 g/ml actinomycin was added to the culture at 0, 30, and 60 min after stimulation with LPS/PMA. Mono Mac 6 cells were collected at each time point and TNF- mRNA was measured by real-time PCR described previously.

Chromatin IP (ChIP) assay for human IRF-3:

Mono Mac 6 cells culture in control or EtOH conditions were plated to 24-well plate at a cell density of 2 10⁶ of cells/ml/well, and stimulated with LPS and PMA for 1 h as stated above. Then the cells were crosslinked with 1% (v/v) formaldehyde at 37C for 20 min. …

Cell transduction with lentivirus and luciferase assay:

… BM cells were transduced overnight by lentivirus with ISRE, AP-1 driving luciferase as a reporter on day 3 before treating EtOH, and followed with EtOH and LPS as described above. After stimulation by LPS/PMA (for Mono Mac 6 cells) or LPS (for BM cells) for 6 h, the cells were harvested and ISRE, AP-1, or TNF- promoters activities were analysis using the Luciferase Reporter Assay System (Promega). The luciferase activities were normalized by total cellular proteins. …

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

Materials:

SNAPtide substrates (Product # 520), Botulinum Neurotoxin Type A (Product #130A) and Botulinum Neurotoxin Type B (Product #136) are products of List Biological Laboratories.

• Product #520 – SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #130A – Botulinum Neurotoxin Type A from Clostridium botulinum
• Product #136 – Botulinum Neurotoxin Type B, Nicked, from Clostridium botulinum

Methods – The above materials were used in the following (refer to poster for details):

• Sandwich ELISA assay showing specificity of BTA for binding to GST-SV2c
•  GST Pull Down Assay
• Dynabead Experiment
• Immobilized Protein G Experiment

Microinjection of Toxin Proteins and Use of Inhibitors:

… LF was purchased from List Laboratories, Inc. …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169L (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

New High Affinity Antibodies Against Botulinum Neurotoxin Type A:

… To test the affinity purified antibodies we determined the titer of the anti-HccA antibodies against Botulinum Neurotoxin Type A (BTA) holotoxin, List Product #130. The ability of the anti-HccA antibodies to capture BTA was also tested using a Sandwich ELISA.

Comparison of FRET Substrates for Botulinum Neurotoxin Type A:

A new FRET substrate for the  zinc endoprotease activity of the 50 kDa light chain (LcA) of the type A Botulinum neurotoxin has been designed and evaluated. This new substrate, which contains the FRET pair Mca/Dnp was compared to the SNAPtide substrate (Product #520 and #521) FRET peptides, which contain the oAbz/Dnp and FITC/DABCYL FRET pairs, respectively …

• Product #130 – Botulinum Neurotoxin Type A from Clostridium botulinum
• Product #520 – SNAPtide Peptide Substrate (o-Abz/Dnp) for C. botulinum Type A Neurotoxin
• Product #521 – SNAPtide Peptide Substrate (FITC/DABCYL) for C. botulinum Type A Neurotoxin

Reagents:

LPS was from List Biological Laboratories (Campbell, CA) …

Results – Monocytes of cHCV patients lack tolerance to TNF -inducing TLR ligands:

… Monocytes of controls and HCV patients were primed in vitro with LPS, a TLR4 ligand, and re-stimulated with LPS to assess homotolerance. In agreement with previous reports (8,9), we found that LPS priming lead to low TNF production in response to a re-challenge with LPS in normal monocytes (Fig 1A). However, homo-tolerance to TLR4 ligand was not found in monocytes from HCV patients (Fig 1B). We further identified that pretreatment with LPS induced hetero-tolerance to subsequent stimulation with TLR2 (PGN), TLR2/TLR6 (PAM₃CSK₄), TLR2/TLR1 (PAM₂CSK), TLR3 (poly I:C) and TLR7/8 (Gardiquimod) ligands in controls (Fig 2A), and in patients with liver inflammation due to non-viral non-alcoholic steatohepatitis (NASH) (Fig 2C) but not in HCV-infected patients (Fig 2B). This data indicated that monocytes of HCV-infected patients have lost TLR tolerance.

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/

Quantification of PA₈₃ bound to chimeras 206 and 264:

Recombinant PA₈₃ (List Biological Laboratories) in 5 mM Hepes, 50 mM NaCl (pH 7.5) was mixed with purified chimeras 206 and 264 in 50 mM Hepes (pH 7.5) in a ratio of 180:1 (equimolar amounts of PA₈₃and VWA domains). Following incubation for 20 min at room temprature, an aliquot from each of the samples was removed and stored at 20 C pending analysis. The remainder of the samples was transferred to an ultracentrifuge tube and underlayed with a 30% (w/w) sucrose cushion in 50 mM Hepes (pH 7.5). Complexes of chimeras decorated with PA₈₃ were pelleted by centrifugation at 200,000g for 45 min. …

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis

…Recombinant B. anthracis lethal factor (rLF) and protective antigen (rPA) were acquired from List Biological Laboratories, Inc. (Campbell, CA) …

Author did not indicate which specific lethal factor was utilized.  List Labs provides Product #172 (Anthrax Lethal Factor (LF), Recombinant from B. anthracis) and Product #169 (Anthrax Lethal Factor (LF-A), Recombinant from B. anthracis Native Sequence).

• Product #171 – Anthrax Protective Antigen (PA), Recombinant from B. anthracis