1. Oxidative Stress, Septic Shock, and Survival Matthew Greenwood April 29, 2015

2. Hypothesis: double blockade of complement C5 and the TLR coreceptor CD14 will improve survival during polymicrobial sepsis

3. Methods Use cecal ligation model to induce polymycrobial sepsis Treat C57BL/6 mice with anti-inflammatory agents – Coversin – Anti-mouse CD14 Monitor – Levels of cytokines and other biomarkers – Mouse survival

4. What is Coversin? Inhibitor of complement C5 – Prevents immune chemotaxis – Prevents formation of membrane attack system Originally from Ornithodoros moubata http://www.vipimmunopharma.com/coversin/ http://lymeaware.free.fr/lyme/Websave/maladie satiques/www.maladies-a- tiques.com/Ornithodoros%20moubata.jpg

5. Figure 1 Cytokine levels after 24 hours

6. Figure 2 Inflammatory marker levels after 24 hours.

7. Figure 3 Inflammatory marker levels after 24 hours.

8. Figure 4 Subject survival

9. Conclusions Combined C5 and CD14 inhibition significantly improves systemic inflammation, clinical signs, and survival rate Reducing the magnitude and duration of both the pro- and anti-inflammatory conditions improves outcome

10. Future directions Correlate cytokine levels with mortality and morbidity Treat with antibiotics with the immunosupressants Testing in humans to improve patient outcomes

11. Hypothesis: protecting mitochondria with mitochondrial targeted antioxidants will alleviate myocardial inflammation and rescue heart function in sepsis

12. Background

13. Methods Used intratracheal injection of Streptococcus pneumonia to induce sepsis Sprague-Dawley male rats served as the model organism Administered Mito-Vit-E in a 21.5 uMol/kg dose

14. Figure 1 Distribution of Mito-Vit-E

15. Figure 2

16. Figure 3 Oxidation of molecules

17. Figure 4 Mitochondrial membrane integrity

18. Figure 5

19. Figure 6

20. Figures 7 & 8

21. Conclusions Mitochondrial targeted Vitamin E – Improved antioxidant capacity – Reduced Oxidation of mitochondrial molecules – Protected mitochondrial structure – Reduced Cytokine production – Reduced neutrophil infiltration – Protected heart function

22. Future Directions Survival studies Administer with antibiotics Testing in humans to improve patient outcomes

23. My Experiment Hypothesis: Administration of reactive oxygen species or pro-oxidative species will improve immune response and survival following sepsis Specific Aim: Determine the reactive oxygen species or oxidants that are able to boost immune response in healthy and immunosuppressed mice to recover from sepsis.

24. Experimental Approach Treat mice with oxidants and mitochondrially targeted antioxidants Determine cytokine levels Find optimal treatment for subject survival