Bacterial Toxin Research Citations

Materials:

… E. coli O111 B4 derived LPS was obtained from List Biological Laboratories, Inc. (Campbell, CA, USA).

Results:

Cox2 and Tnfa inhibition by antioxidants alone and in equimolar combinations. The inhibitory effects of BHTrelated compounds on LPS– or P. gingivalis-fimbria induced expression of the Cox2 or Tnfa genes in RAW264.7 cells were investigated at a non-cytotoxic concentration of 10 M using real-time PCR. LPS-induced gene expression of Cox2 was inhibited more dramatically by the BHT/BHA combination than by either antioxidant alone (Figure 2). The 1:1 BHT/BHA combination induced a 50% decrease in gene expression of Cox2. Similarly, equimolar BHT/TBP and BHA/TBP combinations inhibited the expression of Cox2weakly. LPS-induced expression of Tnfawas weakly but significantly suppressed by the 1:1 BHT/BHA combination, whereas the other antioxidant combinations had no suppressive effect (Figure 3). By contrast, fimbriainduced expression of the Cox2 gene was slightly but significantly inhibited by BHA, BHT and TBP, and by the BHA/TBP combination. The equimolar BHT/BHA and BHT/TBP combinations, particularly the former, induced a 50% decrease in the expression of Cox2(p<0.01; Figure 4). BHA, BHT, and TBP did not inhibit the expression of Tnfa mRNA induced by P. gingivalis fimbriae, whereas each combination, particularly BHT/BHA, induced a 50% decrease in the expression of TnfamRNA (p<0.01; Figure 5). These results indicate that the BHT/BHA combination exerted a strong inhibitory effect on the expression of both Cox2and TnfamRNA. Cox2 and Tnfa inhibition by different combinations of two antioxidants at different molar ratios. The results are shown in Figures 6 and 7, respectively. The inhibitory effects of the BHT/BHA combination on LPS-stimulated expression of the Cox2 and Tnfa genes were investigated using different BHA concentrations and time periods. The BHT/BHA combination at molar ratio of 1:2 and 2:1 inhibited LPS-stimulated gene expression of Cox2 more markedly than did their 1:1 combination. However, treatment with an excess of BHA, as in the 1:3 BHT/BHA combination had no inhibitory effect (Figure 6). LPS induced gene expression of Tnfa was more greatly suppressed by the 1:2 and 2:1 BHT/BHA combinations, than by the 1:1 combination (p<0.05) (Figure 7). The 1:2 and 2:1 BHT/BHA combinations induced a 50% decrease in LPS-induced gene expression of Tnfa. By contrast, treatment with excess BHA in the 1:3 BHT/BHA combination induced a more marked expression of Tnfa gene than did stimulation with P. gingivalisfimbriae alone. These results indicate that a combination of BHT/BHA at 1:2 and 2:1 molar ratios has a more effective antiinflammatory activity than that at 1:1 or of either antioxidant alone. 

Product #201 – LPS from Escherichia coli O111:B4

…Anti cholera toxin B-subunit (anti-CTb), List Biological Laboratories, Inc. #703, …

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

Cells and Reagents:

…Polyclonal goat antibody (#771B) against Protective Antigen (PA) was from List Biological Laboratories (Campbell, CA) and used at 1/2000 dilution…

EAE Model:

Pertussis Toxin (List Biological Laboratories Inc.) was injected 200 ng per mouse in PBS intraperitoneally on days 0 and 2 after MOG 3555 injections. …

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)

Reagents:

…Escherichia coli LPS 0111:B4 obtained from…List Biological Laboratories (Campbell, CA) was used for studies involving human PBMCs and mice,…

Human PBMCs and Cytokine Induction by LPS:

PBMCs from healthy human subjects were prepared under a Hebrew University Institutional Review Boardapproved protocol described elsewhere [3, 8] and were cultured at 1 10⁶ cells/mL in Roswell Park Memorial Institute 1640 medium [3, 8]. Cells were allowed to rest at 37C in 5% CO₂ for 4 hours. After LPS stimulation of cells in the presence or absence of AB103/p2TA, culture medium was collected at intervals for triplicate evaluation of cytokines by an enzyme-linked immunoassay (ELISA) [3, 8].

LPS Challenge:

CD1 or BALB/c mice were administered 20 mg D-galactosamine (D-galN) intraperitoneally, followed in 2 hours by 0.25 mg/kg E. coli 0111:B4 LPS. AB103 was injected intraperitoneally 30 minutes before LPS.

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

Chronic EAE immunization:

Preparations of 200-g MOG₃₅₅₅ peptide emulsified in CFA… were injected subcutaneously into the lower flanks of 8-week-old female C57BL/6J mice (Jackson Laboratory, Bar Harbor, MN). Two intraperitoneal (i.p.) injections of 400-ng pertussis toxin each (no. 181, List Biological Laboratories,…)

• Product #181 – Pertussis Toxin from B. pertussis, Lyophilized (Salt-Free)

ANIMAL EXPERIMENTS:

…At 3 weeks post infection, EAE induction treatment was commenced: mice were immunized via sub-cutaneous injection at two sites with MOG peptide MOG_35-55…, containing 4 mg/ml Mycobacterium tuberculosis H37 Ra… An intraperitoneal injection of 200 ng pertussis toxin (List Biological Laboratories Inc., Campbell, CA, USA) was also administered on the same day, with a second subsequent dose 2 days later. …

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)

Induction, clinical evaluation and treatment protocols of EAE:

… EAE was induced by MOG₃₅₅₅ in female mice used between 8 and 10 weeks of age. Briefly, each mouse was immunized subcutaneously with 300µg of MOG₃₅₅₅ emulsified with an equal volume of complete Freund’s adjuvant…and then injected to the caudal vein with 200ng of pertussis toxin (PTX) (dissolved in 200µl PBS, List Biological Laboratories, USA) at the time of immunization and 2 days later.

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)

Dorsal column transection with sciatic nerve conditioning lesion:

… Five days before tissue collection, the left sciatic nerve was exposed again and 1.5 l of 10 mg/ml cholera toxin subunit (CT; List Biological Laboratories), a trans-ganglionic tracer, was injected into the proximal nerve section using a pulled glass capillary tube pipette.

For immunohistochemical analyses of tracer-labeled regenerated axons, a series of every second section of the T4T9 spinal cord segments (1116 sections per mouse, 100 m apart) was processed for CT immunoreactivity, with the starting section number chosen for each mouse at random. Individual sections were washed,…and incubated in goat anti-CT (List Biological Laboratories, 1:8000) for 15 h.

• Product #104 – Cholera Toxin B Subunit (Choleragenoid) from Vibrio cholerae in Low Salt
• Product #703 – Anti-Cholera Toxin B Subunit (Goat)
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Antibodies, reagents and medium:

…CT was from List Biological Laboratories (Campbell, CA, USA); …

Measurement of CT– and BLG-specific antibodies in mouse serum:

…For the measurement of CT-specific antibodies, Maxisorp microtitre plates (Nunc) were coated with 100 µl/well of 1 µg/ml CT in PBS and incubated overnight at room temperature. …

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