A pivotal role of Rho GTPase in the regulation of morphology and function of dendritic cells

Dendritic cell (DC) is the most potent activator of CD4(+) T cells and has unique dendrites and veils. To explore the function of Rho in DC, exoenzyme C3 from Clostridium botulinum was used as a specific inhibitor of Rho. Tre atment of DC with C3 (DC/C3) resulted in profound morphological changes by losing dendrites and emerging of shrunk membrane processes that were in par allel with marked reduction of polymerized actin in the marginal area. Inac tivation of Rho-associated coiled coil-containing kinase (p160ROCK) by a sp ecific ROCK inhibitor Y-27632 also led to disappearance of dendrites of DC with retaining large membrane expansions. In scanning electron microscopy, untreated DCs interacted with CD4(+) T cells more efficiently than DC/C3. C onjugate formation assay showed that the number of DCs associated with CD4( +) T cells was 2-fold higher in untreated DCs than that of DC/C3. Alloantig en-presenting capacity of DC/C3 was significantly suppressed in a dose-depe ndent manner. Because C3 treatment did not affect the surface expression of HLA, costimulatory, and adhesion molecules of DC, we examined cytokine pro duction of DC and naive CD4(+) T cells to further elucidate the inhibitory mechanism of MLR. Unexpectedly, DC/C3 increased IL-12 production after LPS stimulation. Naive CD4(+) T cells cocultured with DC/C3 produced the increa sed percentage of IFN-gamma -producing cells, whereas the percentage of IL- 2-producing T cells was decreased. These results demonstrate that Rho GTPas e in DC controls both characteristic shape and immunogenic capacity.