Citations

Immunization protocol:

A standard nasal immunization protocol was used in this study [12]. C57BL/6 mice were nasally immunized to both nostrils on days 0, 7 and 14 with 25 g of diphtheria toxoid (DT) (1050 Lfu/mg) kindly provided by the Biken Foundation (Osaka University) either alone or combined with 5g of the LPS-free form of mCT or 0.5g of nCT (List Biologic Laboratories Inc.) as mucosal adjuvants.

Detection of neutralizing Abs by use of diphtheria toxin-specific Vero cell assay:

The cytotoxicity induced by the diphtheria toxin (List Biological Laboratories Inc.) in Vero cell cultures was used to measure the toxin-specific neutralizing titers of the Abs induced by the nasal vaccine.

• Product #100B – Cholera Toxin (AZIDE-FREE) from Vibrio cholerae
• Product #150 – Diphtheria Toxin, Unnicked, from Corynebacterium diphtheriae

Materials:

TeTx was provided by List Biological Laboratories, Inc. (Campbell, CA, USA). …

Results:

• TeTx, like TPA, induces [3H]5-HT uptake inhibition in rat-brain synaptosomes
• TeTx induces dose-dependent translocation of PKC isoforms
• TeTx induces both PLC activation and PLCQ1 phosphorylation at Tyr residues via tyrosine kinase receptors (TyrKRs)
• TeTx, like TPA, induces protein phosphorylations in Ser residues and specic SERT phosphorylation
• Bisindolylmaleimide-1 (BIM-1) blocks the TeTx and TPA induction of SERT phosphorylation

• Product #190 – Tetanus Toxin from Clostridium tetani

Reagents:

LPS from the LCD 25 strain of Escherichia coli K12 (Rb-LPS mutant) and LPS of E. coli 0111:B4 (S-LPS) were purchased from List Biological Laboratories (Campbell, CA).

Microtiter Plate LPS Binding Assay:

Immulon-2 96-well microtiter plates were coated with LCD 25 E. coli K 12 LPS or 0111:B4 E. coli LPS (5 g/well), AP-SP-A (positive control for SP-D binding) and myosin (positive control for SP-A binding [550 ng/well]) or nothing (BSA) in 0.1 M Na₂CO₃, 20 mM EDTA, pH 9.6, for 3 h at 37C. After incubation, the LPS solution was removed, and the plates were air-dried overnight. …

• Product #314 – LPS from Escherichia coli K12 strain LCD25
• Product #201 – Escherichia coli O111:B4

Mice and reagents:

Female C3H/HeJ mice 5 weeks of age were purchased from the Jackson Laboratory (Bar Harbor, Me) and maintained on peanut-free chow under specific pathogen-free conditions. …

Freshly ground whole peanut was used as the antigen. Crude peanut extracts (CPEs), Ara h 1 and Ara h 2, were prepared as described previously.^12, 13 Cholera toxin was purchased from List Biological Laboratories, Inc (Campbell, Calif). …

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