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*
Department of Pediatrics and Clinical Immunology, Mie University School of Medicine, Tsu, Mie, Japan; and
Department of Pediatrics, National Mie Chuo Hospital, Tsu, Mie, Japan
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. Treatment of
DC with C3 (DC/C3) resulted in profound morphological changes by losing
dendrites and emerging of shrunk membrane processes that were in
parallel with marked reduction of polymerized actin in the marginal
area. Inactivation of Rho-associated coiled coil-containing kinase
(p160ROCK) by a specific 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. Conjugate
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. Alloantigen-presenting capacity of DC/C3 was significantly
suppressed in a dose-dependent manner. Because C3 treatment did not
affect the surface expression of HLA, costimulatory, and adhesion
molecules of DC, we examined cytokine production 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 increased percentage of IFN-
-producing cells, whereas
the percentage of IL-2-producing T cells was decreased. These results
demonstrate that Rho GTPase in DC controls both characteristic shape
and immunogenic capacity.
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