Bacterial Toxin Research Citations

Western blot analysis:

Cell lysates and culture supernatants were obtained from cultures at the transition phase of growth (4 h). Four-milliliter culture samples were centrifuged at 10,000 g for 10 min. Preparation of cell lysates for Western blot analysis of AtxA was performed according to the methods described by Hammerstrom et al. (24) … Primary antisera (anti-LF [R. J. Collier] and anti-EF [R. J. Collier]) or antibody (anti-PA [List Biological Laboratories, Inc., Campbell, CA]) was added to TBS-T and allowed to react with the membrane for 1 h at RT. …

Chemicals and Antibodies:

… We used LT from List Biological Laboratories. …

Results-

LT enhances ETEC H10407 adherence to HCT-8 cells:

We showed previously that ETEC strains possessing LT adhere more avidly to cultured intestinal epithelial cells and induce the production of higher levels of cAMP, as compared with ETEC strains lacking LT (Johnson et al., 2009). To begin to determine the mechanism governing LT-induced adherence, we further developed assays using both the prototypical human isolate ETEC H10407 (Evans et al., 1975) and HCT-8 cells, a cell line derived from a human ileocecal colorectal adenocarcinoma (Tompkins et al., 1974) frequently used to study both ETEC … We also performed bacterial adherence assays to verify that LT expression enhances ETEC adherence to HCT-8 cells. Relative to the adherence of eltA ETEC (set to 1.0), wt ETEC adherence was increased 6.7 0.5-fold (Fig. 1B). This difference in adherence between wt and eltA ETEC was independent of the multiplicity of infection (moi) used in infection experiments (not shown). By exposing HCT-8 cells to purified LT holotoxin, we augmented eltA adherence to levels not significantly different (6.0 0.5-fold) from wt ETEC. By contrast, adding purified LT to wt ETEC induced no further increase in adherence. Thus, LT enhances both cAMP production and ETEC adherence to HCT-8 cells.

Lethal Toxin (LT) = LF + PA

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

The PA standard used in these experiments was obtained from List Biological Labs, Inc. and used at 1000 ng/mL concentrations.

Place Exchange of PA onto AuNPs:

For a ligand to peptide feed ratio (L/P) of 25:1 GS/PA, 20 mg GS AuNP was
added to 1.63 mg loop PA or 1.54 mg linear PA in 6.66 mL deionized water and stirred
for 3 days at room temperature.  For 25:1 TioEG/PA, 25 mg TioEG MPC was added to
3.94 mg loop PA or 3.72 mg linear PA in 25 mL deionized H2O at room temperature.

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

Antigens:

… Recombinant rPA from Bacillus anthracis were purchased from List Biological Laboratories, Inc. …

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

Analysis of cytokine and chemokine expression in BMDCs or in nasal septal tissues:

BMDCs were cultured for 5 days as described above. A total of 4 10⁶ BMDCs/treatment were stimulated with 0.001, 0.01, or 0.1% NE, 1, 10, or 30 g/ml CT (List Laboratories). As a positive control BMDCs were treated also with 1 or 10 ng/ml LPS Salmonella minnesota (List Laboratories) or left untreated as negative control. …

• Product #304 – LPS from Salmonella minnesota R595 (Re)

Author did not specify which Cholera toxin was utilized.  List Labs provides Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae and Product #101 – Cholera Toxin from Vibrio cholerae.

Generation of MDDC:

… On day 6, MDDC were retrieved, washed and pulsed with CT (1 µg/ml) (List Biological Laboratories, Inc., Campbell, CA, USA) or mockpulsed in 1ml of medium for 2h at 37°C. …

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

… Cells were stimulated with 100 ng/ml ultrapure LPS from Salmonella minnesota R595 (List Biological Laboratories) in the presence or absence of 100 nM CGRP or CGRP 8-37 CGRP (Bachem) for the indicated time periods. …

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

… Purified LPS was purchased from List Biologicals (San Jose, CA, USA). …

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/

Toxicity assays in vitro:

For toxicity studies, indicated amounts of ssRNA40 complexed with LyoVec, imiquimod, loxoribine (all from InvivoGen, San Diego, CA), LPS (List Biological Laboratories, Campbell, CA),and other reagents were added to cell cultures for indicated durations. …

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/

Reagents:

The reagents used in this study were Escherichia coli serotype 0111:B4 LPS (List Biological Laboratories, Campell, CA …

Cell Culture:

Bone marrowderived macrophages (BMDMs) were cultured in macrophage medium (RPMI 1640, 10% FBS, 30% L929 cell supernatant, 2 mM l-glutamine, 100 IU/mL penicillin, and 100 g/ml streptomycin) as described (20). After being cultured in macrophage medium for 6 days, BMDMs were isolated, counted, and cultured in macrophage media for 24 hours and then stimulated with LPS (10 or 100 ng/ml), IL-4/IL-13 (10 ng/ml), IL-10 (10 ng/ml), or RPMI 1640 for up to 48 hours. Alveolar macrophages isolated with repeated BAL using PBS were cultured for 24 hours in macrophage media and then stimulated with LPS for 4 hours. BEAS-2B cells were cultured in RPMI 1640 supplemented with 10% BSA, penicillin, and streptomycin for 5 to 6 days until they reached 90% confluence. The culture medium was then replaced, and low-molecular-weight heparin, syndecan-4, or RPMI 1640 was added for 1 hour. Cells were washed with PBS and incubated with LPS (1,000 ng/ml), TNF- (1 ng/ml), or RPMI 1640 without heparin or syndecan-4 for 3 hours. Mouse tracheal airliquid interface cultures were performed as described and stimulated with LPS or RPMI 1640 for 4 and 24 hours (33). Supernatants were removed and RNA was harvested.

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

… PTX was purchased from List Laboratories (Campbell, CA, USA). …

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)