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The Journal of Immunology, 2001, 167: 3585-3591.
Copyright © 2001 by The American Association of Immunologists

A Pivotal Role of Rho GTPase in the Regulation of Morphology and Function of Dendritic Cells1

Michihiro Kobayashi*, Eiichi Azuma2,*, Masaru Ido*, Masahiro Hirayama*, Qi Jiang*, Shotaro Iwamoto*, Tadashi Kumamoto*, Hatsumi Yamamoto{dagger}, Minoru Sakurai* and Yoshihiro Komada*

* Department of Pediatrics and Clinical Immunology, Mie University School of Medicine, Tsu, Mie, Japan; and {dagger} 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-{gamma}-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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