Figure 7.

Induction of acute inflammatory responses by wild-type TcdB in the presence of a cysteine protease inhibitor. TcdB with or without AWP19, or AWP19 alone was injected into ligated ileal loops. The mice (n = 6) were killed and the ileal loops were collected for subsequent analysis. (A) Immunoprecipitation of the GTD fragment in tissue lysates. Mouse ligated ileal loops were injected with TcdB alone or TcdB in the presence of AWP19 and the intestinal tissues were collected 4 hours later for the immunoprecipitation assay. Samples from 3 different mice treated with TcdB together with AWP19 were shown. Wild-type TcdB pretreated with InsP₆ was loaded as an immunoblotting control. (B–E) Analysis of ileal loops by (B) quantitation of intestinal fluid accumulation by weight-to-length ratio, (C) MPO activity, (D) messenger RNA expression of inflammatory cytokines by reverse-transcription PCR, and (E) H&E-stained intestinal tissue sections are shown. *P < .05, **P < .01 (compared with PBS control). GAPDH, glyceraldehyde-3-phosphate dehydrogenase; KC, keratinocyte chemoattractant.