EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions •Repeat EAEC40 with nourished control diet made with egg white protein •Generate additional data for Bolick, et al. 2013 manuscript Day mice delivered -3 Tues - 14 Tues Design 1) C57Bl/6 male mice 4 weeks old on arrival 2) Nourished (N) or Nourished-Zinc (NZD) diet fed for two weeks 3) Antibiotic cocktail pretreatment for 2 days, followed by 1 day off 4) weights, stools collected every day, sac day 3 EAEC 10 9 0 weights and stools Diet Start -1 Thur s ab Euthanasia for collection 3 Zinc depletion

EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

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EAEC51 Study start 8-13-2013N or NZD fed 14d/ab pretreatment/10^9 EAEC 042

EAEC51 Study DesignObjectives/Questions•Repeat EAEC40 with nourished control diet made with egg white protein•Generate additional data for Bolick, et al. 2013 manuscript

Day

mice delivered

-3

Tues

-14

Tues

Design1) C57Bl/6 male mice 4 weeks old on arrival2) Nourished (N) or Nourished-Zinc (NZD) diet fed for two weeks3) Antibiotic cocktail pretreatment for 2 days, followed by 1 day off4) weights, stools collected every day, sac day 3

EAEC 109

0weights and stools

Diet Start

-1

Thurs

Euthanasia for collection

3Zinc depletion

Page 2: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

Page 3: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

Page 4: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

Page 5: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

To do for EAEC 51 Mx

• Histology (done, needs pics and IHC for CD45)• qPCR for cytokines• qPCR for in vivo virulence factor expression• Final revisions and where to submit

Page 6: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

IL-17r Study

Page 7: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

Page 8: EAEC51 Study start 8-13-2013 N or NZD fed 14d/ab pretreatment/10^9 EAEC 042 EAEC51 Study Design Objectives/Questions Repeat EAEC40 with nourished control

IL-17rKO To Do

• Histology (done, needs pics and IHC?)• qPCR (Th17 pathway plate)• Tissue burden (done, needs graph)• Mice are in, MTA is still in approval process• IL-25?