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Autophagy is induced by the type III secretion system of Vibrio alginolyticus in several mammalian cell lines

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Abstract

Vibrio alginolyticus is a gram-negative bacterium and has been recognized as an opportunistic pathogen in marine animals as well as humans. Here, we further characterized a cell death mechanism caused by this bacterium in several mammalian cell lines. The T3SS of V. alginolyticus killed HeLa cells by a very similar cell cytolysis mechanism in fish cells, as evidenced by cell rounding and LDH release; however, DNA fragmentation was not observed. Further studies showed that caspase-1 and caspase-3 were not activated during the T3SS-mediated cell death, indicating that the death mechanism is completely independent of pyroptosis and apoptosis in HeLa cells. Conversely, autophagy was detected during the T3SS-mediated cell death by the appearance of MDC-labeled punctate fluorescence and accumulation of autophagic vesicles. Moreover, western blot analysis revealed increase in conversion of LC3-I to LC3-II in infected mammalian cell lines, confirming that autophagy occurs during the process. Together, these data demonstrate that the death process used by V. alginolyticus in mammalian cells is different from that in fish cells, including induction of autophagy, cell rounding and osmotic lysis. This study provides some evidences hinting that differences in death mechanism in responses to V. alginolyticus infection may be attributed to the species of infected cells from which it was derived.

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Abbreviations

CLC:

Carp leucocyte cell

DMEM:

Dulbecco’s minimal essential medium

FHM:

Fathead minnow epithelial cell

EPC:

Epithelioma papulosum cyprinid

FBS:

Fetal bovine serum

LC3:

Microtubule-associated protein light chain 3

LDH:

Lactate dehydrogenase

PFA:

Paraformaldehyde

PBS:

Phosphate-buffered saline

MDC:

Monodansylcadaverine

MEM:

Minimum essential medium with Earle’s balanced salts

SDS–PAGE:

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

STS:

Staurosporine

T3SS:

The type III secretion system

TUNEL:

Terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling

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Acknowledgments

We thank Dr. Luo Xiao-Chun (SCUT, CHINA) for providing mammalian cell lines. This research was supported by grants from the National Basic Research Program of China (2006CB101803), the National Key Technology R&D Program (2009BAB44B02), the National Natural Science Foundation of China (30901117) and the China Postdoctoral Science Foundation (20090450100 and 201003364). The authors also gratefully acknowledged the support of the K.C. Wong Education Foundation, Hong Kong.

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Authors and Affiliations

  1. Key Laboratory of Applied Marine Biology of Guangdong Province (LAMB) and Key Laboratory of Marine Bio-Resources Sustainable Utilization (LMB), South China Sea Institute of Oceanology, Chinese Academy of Sciences, Guangzhou, 510301, People’s Republic of China

    Zhe Zhao, Lvping Zhang, Chunhua Ren, Jingjing Zhao, Chang Chen, Xiao Jiang, Peng Luo & Chao-Qun Hu

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  1. Zhe Zhao
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  2. Lvping Zhang
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  3. Chunhua Ren
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  4. Jingjing Zhao
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  5. Chang Chen
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  6. Xiao Jiang
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  7. Peng Luo
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  8. Chao-Qun Hu
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Corresponding author

Correspondence to Chao-Qun Hu.

Additional information

Communicated by David Kelly.

Z. Zhao, L. Zhang—contributed equally to this work.

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Zhao, Z., Zhang, L., Ren, C. et al. Autophagy is induced by the type III secretion system of Vibrio alginolyticus in several mammalian cell lines. Arch Microbiol 193, 53–61 (2011). https://doi.org/10.1007/s00203-010-0646-9

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  • DOI: https://doi.org/10.1007/s00203-010-0646-9

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