Noteworthy is the fact that we now have proven that C. jejuni infection of cells results in B1 integrin and EGF receptor association and EGF receptor activation. So, we particularly examined if MBCD therapy of HeLa cells disrupts B1 in tegrin and EGF receptor association. The results of this experiment revealed that MBCD totally inhibited EGF receptor activation and disrupted the association on the B1 integrin and EGF receptor, as judged by IP ex periments. HPBCD treatment method of HeLa cells also inhibited EGF receptor activation and disrupted the association from the B1 integrin and EGF receptor, and MBCD treatment of Caco 2 cells prevented EGF recep tor activation. These findings present a plausible explanation to the inhibi tory effect of MBCD on C. jejuni internalization.
Discussion Caveolae are flask like structures which are enriched in cholesterol and glycosphingolipids, on the other hand, caveolae are various from planar lipid rafts according to the presence in the cholesterol binding more info here protein caveolin 1. Caveolin one dose dependent manner. Wooldridge et al. observed a dose dependent reduction in C. jejuni invasion of Caco 2 cells when the cells were handled with filipin III, and concluded that caveolae are involved with C. jejuni uptake. Subsequently, Watson and Galan re ported the therapy of human T84 cells with MBCD blocked C. jejuni internalization. On the other hand, determined by supplemental assays, Watson and Galan speculated that C. jejuni internalization might not be associated with caveolae mediated endocytosis, but that that caveo lae or caveolin one could selleck chemical NSC 74859 perform a part within the host cell signal ing occasions vital for bacterial uptake.
Our data indicate that C. jejuni internalization takes place in the caveolae independent manner. We did discover that deal with ment of HeLa cells with MBCD and HPBCD, two cholesterol depleting compounds, reduced C. jejuni in ternalization within a dose dependent method. These results support the hypothesis that C. jejuni internalization oc curs within a caveolae dependent manner, as caveolin 1 is a cholesterol binding protein along with the treatment method of cells using a cholesterol depleting compound would disrupt the cellular localization of caveolin one. However, knock down of caveolin one protein in HeLa cells by therapy with caveolin 1 siRNA had no impact on C. jejuni intern alization. In addition, therapy of Caco 2 cells with MBCD inhibited C. jejuni internalization. Caco 2 cells tend not to express caveolin one and, as a consequence, do not have caveolae. We expressed caveolin 1 in Caco two cells to attempt to enrich the invasiveness of C. jejuni, but did not observe a modify in the quantity of seems to stabilize the invaginated framework of caveolae.