Transcript The Future of Topical Nasal Therapy for Chronic Rhinosinusitis

Eotaxin: a Potential Target for
Therapy in Chronic Rhinosinusitis
Alisha N. West, MD
Grand Rounds
University of California, Los Angeles
December 1, 2010
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
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
Hypothesis
Eotaxin plays a role in CRS
inflammation and could serve as a
potential target for therapy
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
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
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
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
Tissue Harvest
Results
H&E Stain 10x
CRS
Control
Eosinophilia
CRS
Control
Eosinophilia in Endosinus
Mucosa
5
4.5
4
3.5
3
2.5
2
1.5
1
0.5
0
Normal
CRS
Normal
CRS
Eotaxin ELISA
20
18
16
14
12
10
8
6
4
2
0
O hr
24 Hr
NS TSB
NS
Pseud
NS
CS TSB
CS
Staph
Pseud
CS
Staph
Luminex Results
120
100
80
Concentration in
pg/ml
60
40
20
0
IL1a
IL4
IL6
IL8
IL12p40IL12p70 TNFa GMCSFRANTES
IL13
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
References
1. Ponikau JU, Sherris DA, Kephart GM, Kern EB, Congdon DJ, Adolphson CR,
Springett MJ, Gleich GJ, Kita H.
Striking deposit ion of toxic eosinophil major basic protein in mucus : implications for
chronic rhinosinusitis. J Allergy Clin Immunol. 2005 Aug;116(2):362-9.
2. Dhong HJ, Kim HY, Cho DY.
Histopat hologic characterist ics of chronic sinusitis with bronchial asthma.
Acta Otolaryngol. 2005 Feb;125(2):169-76.
3. Sasama J, Sherris DA, Shin SH, Kephart GM, Kern EB, Ponikau JU. New paradigm
for the roles of fungi and eosinophils in chronic rhinosinusit is.
Curr Opin Otolaryngol Head Neck Surg. 2005 Feb;13(1):2-8. Review.
4. Bernardes JF, Shan J, Tewfik M, Hamid Q, Frenkiel S, Eidelman DH.
Prot ein nitration in chronic sinusit is and nasal polyposis: role of eosinophils.
Otolaryngol Head Neck Surg. 2004 Nov;131(5):696-703.
5. Seto H, Su zaki H, Shioda S.
Immunohistochemical localizat ion of eotaxin immunoreact ivity in nasal polyps.
Acta Otolaryngol Suppl. 2004 Aug;(553):99-104.
6. Ferguson BJ.
Categorization of eosinophilic chronic rhinosinusitis.
Curr Opin Otolaryngol Head Neck Surg. 2004 Jun;12(3):237-42. Review.
PMID: 15167036[P ubMed - indexed for MEDLINE]
Thank You