1. Vibrio parahaemolyticus Disruption of Epithelial Cell Tight Junctions Occurs Independently of Toxin Production Tarah Lynch, Scott Livingstone, Enrico Buenaventura, Erika Lutter, Jason Fedwick, Andre G. Buret, David Graham, and Rebekah DeVinney INFECTION AND IMMUNITY, Mar. 2005, p. 1275–1283

2. Background Vibrio parahaemolyticus Toxin production: TDH(thermostable direct hemolysin) & TRH(TDH-related hemolysin) Barrier function is maintained by tight junction Tight junction comprise of transmembrane proteins (ex: actin 、 occludin 、 ZO-1) Cytoskeletal components are anchored to tight junction

3. Background Latex agglutination test

4. Background Lactate dehydrogenase (LDH)

5. Background TER ( transepithelial resistance ) & paracellular permeability

6. Background Southern confirmation

7. -/++/--/-+/+ PCRSouthern confirmation TLH TRH TDH TDH / TRH

8. +/+ +/- -/+ -/-

9. Purified TDH Uninfected TDH(-) / TRH(-) toxigenic nontoxigenic -/+ +/- +/+ +/- -/- V. parahaemolyticus disrupts TER and that this disruption occurs independently of toxin production.

10. +/+ +/- -/+ -/- TER ↓  paracellular permeability ↑ V. parahaemolyticus disrupts TER and that this disruption occurs independently of toxin production.

11. -/- +/+ -/- red/green V. parahaemolyticus disrupts TER and that this disruption occurs independently of toxin production.

12. +/+ +/- -/+ -/- White ： 3 hr Black ： 6 hr Disruption of epithelial barrier function does not correlate with cytotoxicity V. parahaemolyticus disrupts TER and that this disruption occurs independently of toxin production.

14. Discussion 破壞上皮的 barrier function 產物 TDH & TRH 不 是主要的腳色，可能有其他未知物質 TDH/TRH TDH TRH + + + TDH purify TER ↓

15. Discussion 高濃度 TDH(> 5 HU/ml) 一樣會造成 TER↓ ，並增加 LDH release ，所以並不排除 TDH and/or TRH 的毒性，只是作用較晚 非 TDH and/or TRH 的毒素，影響上皮的 barrier function 作用較早 (1.5 ~ 3 hr postinfection)

16. Discussion 不同的 toxin genotypes 腸炎弧菌，在上皮 的 barrier function 有相同的反應，說明可 能是由其他物質造成的 當測 LDH release 指出破壞上皮的 barrier function 跟 cytotoxicity 無關， LDH release 測得於 cell lysis ，但不能決定 cell 開始 apoptosis

17. TDH/TRH TDH TRH + + + TDH purify TER ↓ V. parahaemolyticus disrupts TER and that this disruption occurs independently of toxin production. Summary

18. TER ↓ TDH purifyLDH release Disruption of epithelial barrier function does not correlate with cytotoxicity Summary