Fil: Gutierrez, Maximiliano Gabriel. Laboratorio de Biología Celular y Molecular, Instituto de Histología y Embriología–Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza; Argentina.Fil: Saka, Hector A. Departamento de Bioquímica Clínica, Centro de Investigaciones en Bioquímica Clínica e Inmunología–CONICET, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba; Argentina.Fil: Chinen, Isabel. ANLIS Dr.C.G.Malbrán. Instituto Nacional de Enfermedades Infecciosas. Departamento de Bacteriología. Servicio Fisiopatogenia; Argentina.Fil: Zoppino, Felipe C M. Laboratorio de Biología Celular y Molecular, Instituto de Histología y Embriología–Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza; Argentina.Fil: Yoshimori, Tamotsu. Department of Cell Genetics, National Institute of Genetics, Yata 1111 Mishima; Japan.Fil: Bocco, Jose Luis. Departamento de Bioquímica Clínica, Centro de Investigaciones en Bioquímica Clínica e Inmunología–CONICET, Facultad de Ciencias Químicas, Universidad Nacional de Córdoba, Argentina.Fil: Colombo, María Isabel. Laboratorio de Biología Celular y Molecular, Instituto de Histología y Embriología–Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Facultad de Ciencias Médicas, Universidad Nacional de Cuyo, Mendoza; Argentina.Autophagy is the unique, regulated mechanism for the degradation of organelles. This intracellular process acts as a prosurvival pathway during cell starvation or stress and is also involved in cellular response against specific bacterial infections. Vibrio cholerae is a noninvasive intestinal pathogen that has been studied extensively as the causative agent of the human disease cholera. V. cholerae illness is produced primarily through the expression of a potent toxin (cholera toxin) within the human intestine. Besides cholera toxin, this bacterium secretes a hemolytic exotoxin termed V. cholerae cytolysin (VCC) that causes extensive vacuolation in epithelial cells. In this work, we explored the relationship between the vacuolation caused by VCC and the autophagic pathway. Treatment of cells with VCC increased the punctate distribution of LC3, a feature indicative of autophagosome formation. Moreover, VCC-induced vacuoles colocalized with LC3 in several cell lines, including human intestinal Caco-2 cells, indicating the interaction of the large vacuoles with autophagic vesicles. Electron microscopy analysis confirmed that the vacuoles caused by VCC presented hallmarks of autophagosomes. Additionally, biochemical evidence demonstrated the degradative nature of the VCC-generated vacuoles. Interestingly, autophagy inhibition resulted in decreased survival of Caco-2 cells upon VCC intoxication. Also, VCC failed to induce vacuolization in Atg5-/- cells, and the survival response of these cells against the toxin was dramatically impaired. These results demonstrate that autophagy acts as a cellular defense pathway against secreted bacterial toxins.