1. Cytokine and chemokine responses as predictive markers for distinct stages of Alveolar Echinococcosis
X. Huang1, C.J. Lechner1, B. Grüner2,
W.H. Hoffmann1, P. Kern2, P.T. Soboslay1
1Institute for Tropical Medicine, University Clinics of Tübingen
2Division of Infectious Diseases and Clinical Immunology, University Clinics of Ulm
Innovation for the management of Echinococcosis. 27th March,2014 1

2. Study Objective
Evaluation of cytokines and chemokines in Alveolar Echinococcosis (AE) patients for their use as predictive markers of regression or progression of AE.
Focus of this study is on mediators of inflammation (IL-27, IL-31 and IL-33), as well as chemokines which activate eosinophil granulocytes (Eotaxin-1, Eotaxin-2 and Eotaxin-3).
Chemokine genes expression profiles in AE patients
Eotaxins can activate and chemoattract eosinophil granulocytes and thus drive Th2 type immunity, which predominates in AE patients with active disease 2

3. Sample Collections and Methods
Peripheral blood mononuclear cells (PBMC) from AE patients with cured (n=12), stable (n=36) or progressive (n=21) diseases and infection-free controls (n=10)
PBMC stimulation with E.m. specific antigens
ELISA based quantification of cytokines and chemokines in cell culture supernatants
Gene Microarrays to study the inducible expression of chemokine genes in PBMC from progressive and cured AE patients.

4. Production of IL-31 and IL-33 were depressed in AE patients
Results
Baseline cellular production of
IL-27, IL-31 and IL-33 in AE patients
(Cured:n=10; Stable n=10; Progressive: n=6; CTRL: n=6)
Production of IL-31 and IL-33
were depressed in AE patients

5. Results Baseline cellular production of
Eotaxin-1, Eotaxin-2 and Eotaxin-3 in AE patients
(Cured:n=10; Stable n=10; Progressive: n=6; CTRL: n=6)
Production of Eotaxin-1, Eotaxin-2 and Eotaxin-3 enhanced with AE progression in patients

6. Results
E.m. metacestode antigen-induced production of Eotaxin-2 by PBMC in AE patients
Eotaxin-2 production became strongly activated
by E.m. antigen in cured AE patients

7. Gene Microarrays were applied to study the inducible expression of chemokine genes
Peripheral blood mononuclear cells from patients with progressive and cured AE were stimulated in vitro with E. multilocularis metacestode Antigen or remained without stimulation.
Gene expression is indicated as stimulation index
[Stimulation Index = ]

8. Results The strongest chemokine gene expression
is Liver and Activation Regulated Chemokine (LARC)
in Progressive AE patients 8

9. Results
The highest chemokine gene expression is Monocyte Chemo-attractant Protein 4 (MCP4) in Cured AE patients 9

10. Summary
Cellular production of IL-31 and IL-33 were depressed in progressive AE patients; while Eotaxin-1, Eotaxin-2 and Eotaxin-3 levels were enhanced with active and progressive AE.
E.m. antigen-specific production of Eotaxin-2 was highly inducible in AE patients.
Liver and activation regulated chemokine (LARC) gene expression was highest in progressive AE; while monocyte chemo-attractant protein 4 (MCP4) was strongly expressed in cured AE.
Plasma levels of IL-17B is highest in progressive AE and high in all AE patients. Serum level of IL-12 and IL-13 is low in AE patients.
Spontaneous releases of Pro-inflammatory MIP-1 alpha/CCL3, MIP-1 beta/CCL4, RANTES/CCL5 are higher in AE patients, and persist after Em antigen stimulation, may contribute to prevent tissue damage.
Spontaneous releases of Monokine induced by gamma-inteferon MIG/CXCL9, Th1 type IL-12 and TNF-alpha are low, and persist after Em antigen stimulation.
Spontaneous release of MCP4, tissue and activation associatetd chemokines LARC/CCL20, PARC/CCL18 and TARC/CCL17 are low in AE patients
IL-31 and the Eotaxin chemokines as well as MCP4/CCL13 and LARC/CCL20 are suggested as biomarkers for staging of Alveolar Echinococcosis 10

11. Acknowledgements University Clinics of Ulm Prof. Dr. Peter Kern
Dr. Beate Grüner
University Clinics of Tübingen
Prof. Dr. Peter Soboslay
Dr. Christian Lechner 11

12. Thank you for your attention!
Merci beaucoup