1. Eotaxin: a Potential Target for Therapy in Chronic Rhinosinusitis
Alisha N. West, MD
Grand Rounds
University of California, Los Angeles
December 1, 2010

2. History
Chronic Rhinosinusitis (CRS) is characterized by signs and symptoms of inflammation
Stimulated by environmental pathogen
Chronic epithelial stimulation  immune response with a subset of patients exhibiting eosinophilia
Lack of effective mucociliary clearance
Currently, paucity of effective therapies

3. History
Chronic Rhinosinusitis is one of the leading causes of annual visits to the Otolaryngologist
66 million adults in the US reported sinus problems last year
$5.8 billion dollars is spent annually on treatment for CRS
Symptoms usually recur within three months of therapy

4. Hypothesis
Eotaxin plays a role in CRS inflammation and could serve as a potential target for therapy

5. Background
Eotaxin-1 and 2 implicated in inflammation seen in ABPA and asthma
IL-13 and IL-4 up-regulate production of Eotaxin by epithelial cells
Eotaxin is a chemotactic factor that attracts eosinophils to the site of inflammation
Eosinophils release MBP which has been shown to cause epithelial cell damage in ABPA patients
Toxic levels of MBP have been found in the mucus of CRS patients
Prior to our current project, the role of eosinophils and eotaxin had not been examined in CRS

6. Materials and Methods UNC IRB approved protocols
Sinonasal mucosa harvested during endoscopic sinus surgery60 subjects
Tissue embedded in parrifin and H&E stain performed for EOS
Cell culture protocol
Cellular disaggregation
Plated in serum-free media until confluent, dispersion in trypsin
Re-plated on Milicells™ in air-liquid interface media at density of 250,000 cells/Millicell™
After confluent monolayeraspirate apical media
Washed and re-fed basally for 21 days until ciliated and mucus production
4 cell cultures/specimen
Apical challenge with Pseudomonas, Staph, and TSB
Basal Media collected, washed and replaced daily 0-96 hours

7. Materials and Methods Luminex 22-plex battery of Cytokines
IL-1beta
IL-2
IL-4
IL-5
IL-6
IL-10
IL-12p70
TNF-alpha
IFN-gamma
G-CSF
IP-10
GM-CSF
IL-9
IL-13 KC
MCP-1
MIP-1a
RANTES
IL-1a
IL-7
IL-15
IL-17

8. Materials and Methods
Alveolar Lavage has demonstrated eosinophilia in ABPA and asthma patients as well as animal models
Endosinus lavage with evaluation of eosinophilia in CRS patients and normal volunteers
Complete cell count and Eosinophil count performed with cytospin

9. Tissue Harvest

10. Results

11. H&E Stain 10x CRS Control

12. Eosinophilia CRS Control

13. Eosinophilia in Endosinus Mucosa

14. Eotaxin ELISA

15. Luminex Results

16. Conclusions
There is significant Eosinophilia in CRS endosinus epithelium when compared to normal controls
CRS epithelium produces a higher concentration of eotaxin at baseline and after an infectious challenge when compared to controls
Eotaxin should be investigated as a potential target for therapy in CRS

17. References

18. Thank You