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Archives of Microbiology

, Volume 193, Issue 1, pp 53–61 | Cite as

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

  • Zhe Zhao
  • Lvping Zhang
  • Chunhua Ren
  • Jingjing Zhao
  • Chang Chen
  • Xiao Jiang
  • Peng Luo
  • Chao-Qun HuEmail author
Original Paper

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.

Keywords

Vibrio alginolyticus The type III secretion system (T3SS) Cell death Autophagy Apoptosis 

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

Notes

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.

Supplementary material

203_2010_646_MOESM1_ESM.doc (11.8 mb)
Supplementary material 1 (DOC 12104 kb)

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Copyright information

© Springer-Verlag 2010

Authors and Affiliations

  • Zhe Zhao
    • 1
  • Lvping Zhang
    • 1
  • Chunhua Ren
    • 1
  • Jingjing Zhao
    • 1
  • Chang Chen
    • 1
  • Xiao Jiang
    • 1
  • Peng Luo
    • 1
  • Chao-Qun Hu
    • 1
    Email author
  1. 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 SciencesGuangzhouPeople’s Republic of China

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