Citations

Materials:

… Shiga toxin Stx2 was purchased from List Biological Laboratories, Inc. (Campbell, CA). …

Milk Inhibits Ricin but Not Shiga Stx2 Toxin:

To demonstrate that inhibition of toxin activity is specific to ricin, we also evaluated the effect of milk on the Stx2 produced by enterohemorrhagic strain Escherichia coli O157:H7. This bacterial toxin acts similarly to ricin in inactivating ribosome proteins, and similar to ricin, is composed of two subunits. However, in contrast to ricin, the Stx2 B chain binds specifically to the cell surface receptor glycolipid globotriasylceramides Gb3 and Gb4. In addition, the two toxins seem to use different endocytic pathways (21).
To test the specificity of milk inhibition and antibody neutralization, we compared the inhibition of GFP expression in Vero cells by Stx2 (1 ng/ml) and ricin (1 ng/ml) in the presence and absence of milk or antibodies. Fig. 6 shows that milk had the same protective effect in inhibiting ricin as did the anti-ricin antibody, as indicated by GFP fluorescence intensity levels. The results were nearly the same for the milk samples with and without ricin, indicating that 1% milk was fully effective at inhibiting ricin toxicity. 

List Labs has discontinued Product #162 (Lyophilized) and has replaced Product #162 with Product #162L (Liquid):

• Product #162L (Liquid) – Shiga Toxin 2 from Escherichia coli

Sensory tracing:

Three days prior to sacrifice, all 17 remaining animals in Experiment 1 and the remaining 19 animals in Experiment 2 that did not receive CST tracing, were reanesthetized with isoflurane and 2% O₂, and the sciatic nerve was isolated, crushed, and injected with 5L of cholera-toxin beta (CT; ListBiological Laboratories, Inc., Campbell, CA). The needle was left in place briefly before the skin was closed with wound clips. The animals were given buprenorphine and gentamicin as above.

Tissue collection and histology:

… CT-labeled sensory axons originating from L3L5 were identified by immunohistochemistry for CT followed by ABC staining. An entire set of slides containing all cases from Experiments 1 and 2 that received injections of CT were processed at the same time. Tissue sections were washed with Tris-buffered saline with 2% Triton (TBS-T), treated with 2:1 chloroform:methanol to remove lipids for 1h, rewashed, and treated with 10% methanol, then rinsed with 30% H₂O₂ in distilled H₂O for 1h to quench endogenous peroxidase activity. The tissue was then washed with TBS-T and blocked overnight with 5% normal rabbit serum. This was followed by incubation with goat anti-CT (1:20:000; List Biological Laboratories, Campbell, CA) for 60h, …

Author did not specify which CTB was utilized.  List Labs provides Product #103B (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae)  and Product #104 (Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt).

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

… LPS(lipopolysaccharide) List Biological …

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/

Surgical Procedure:

… Injections were made with an adapted Hamilton syringe of 10 µl, the plunger of which was moved by a programmable microsyringe pump (flow 200 nl/min), connected by way of thin tubing (inner diameter, 0.1 mm) in which the movement of an air bubble was monitored to enable control of the injected volume. The injectate contained a mixture of 1 part 1% CTb (low salt; List Biological Laboratories, 1% w/v in 0.2 M phosphate buffer, pH 7.4) and 4 parts RV (Ruigrok et al., 2008; Prevosto et al., 2009). …

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

Induction of experimental autoimmune encephalomyelitis (EAE):

EAE was induced in mice by active immunization with MOG_35-55 peptide (MEVGWYRSPFSRVVHLYRNGK), of murine origin (W. M. Keck Biotechnology Resource Center, Yale University), as described [25]; following Animal Care and Use Guidelines of the University of Connecticut Health Center (Animal Welfare Assurance # A3471-01). Briefly, on day 0, one group of female mice 7-9 weeks of age was injected subcutaneously with 300 µg of MOG peptide in complete Freunds adjuvant (CFA, DIFCO) into the right and left flank, 100 l per site. These mice were also injected i.p. with 500 ng pertussis toxin (PTX, List Laboratories, Campbell CA) in PBS on days 0 and 2 following the first immunization (referred to as the MOG-CFA/PTX group). The second group of age-matched mice received CFA alone and PTX (500 ng) injections on day 0 and a second injection of 500 ng PTX alone on day 2 (referred to as the CFA/PTX group).

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)

Preparation of Murabutide, Cholera Toxin and Gardiquimod:

… Cholera toxin (List Biological Laboratories, Inc., Campbell, CA) …

Mouse Vaccination:

… In the first study, mice were vaccinated intranasally on day 0 and 21, as previously described [8] for liquid formulations containing 25 µg VLP alone, 25 µg VLP +25 µg MB, 25 µg VLP +100 µg MB, 25 µg VLP +250 µg MB, 250 µg MB alone, 25 µg VLP +1 µg CT (1 mg/ml in PBS, List Biological Laboratories, Inc.) which is a well characterized mucosal adjuvant, …

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

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.