Development of Safe and Non-Self-Immunogenic Mucosal Adjuvant by Recombinant Fusion of Cholera Toxin A1 Subunit with Protein Transduction Domain

Figure 5

In vitro and in vivo toxicities of TCTA1T. (a) 20 μg of TCTA1T and TmCTA1T or 1 μg of CT in 10 μl of PBS was injected into the hind paw. The thickness of the footpad was measured after 24 hrs. (b) Mice were anesthetized with ketamine, and abdomen was opened to make a loop in the small intestine. Then, 10 μg of TCTA1T and TmCTA1T, or 2 μg of CT, in 100 μl of PBS were injected into the loop. The ratio of weight to length was determined after 6 hrs and the data expressed as milligrams per centimeter (mg/cm) of fluid accumulation. (c) BHK21 cells were seeded at 1 × 10⁶ cells/well in a 6-well plate. The cells were incubated with 10 μg of TCTA1T and TmCTA1T, or 1 μg of CT for 3 hrs. The concentration of cAMP in the supernatant was measured by ELISA according to the manufacturer’s instructions. The results are expressed as mean ± SD for the group ( to 4). Data are representative of two separate experiments. The different letters within the same samples are significantly different ().

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