Mye-EUNITER
 

Hrckova, Gabriela

Models

Parasitic infections remain a worldwide human and animal health concern. Cestode parasites such as Echinococcus granulosus, Echinococcus multilocularis, Taenia saginata and Taenia solium are agents of severe human and animal diseases. Belonging to the same group of parasitic helminths, Mesocestoides vogae constitutes an interesting model system to study molecular and immunological mechanisms regulating host–parasite relationship which is in long-term under control of these flatworms. In addition, this cestode species was proposed by WHO (1996) as a suitable model for the slower developing metacestode infections such as Echinococcus spp. and Taenia spp. in pharmacological research.

Major cell types studies and key scientific question(s)

MDSC are key players in pathology and chronicity of infections with tissue-dwelling helminth parasites, of which macrophages, neutrophils and NK cells are dominant populations localized in the close vicinity of larvae in the host´s tissues and peritoneal cavity.
- We will evaluate phenotypes, functions and suppressive capacity of above mentioned myeloid cells isolated from two anatomically distinct infection sites: the liver and the peritoneal cavity (PC) in Balb/c mice infected orally with Mesocestoides vogae larvae.
- We will characterise effector functions of subpopulations of macrophages/ Kupffer cells, neutrophils isolated from liver granulomas and PC in the early and late stage of infection. The study will involve regulation of oxidative stress and impaired cytotoxic activity of granulocytes towards larvae. The rationale for this is that liver granulomatous inflammation and fibrosis interfere with the effects of drugs and ROS produced by MDSC are one of the main triggers of fibrosis.
- We wish to understand mechanisms how imunogenic molecules secreted by larvae (collected after in vitro culturing ) affect functions, receptor expression and transcription factors of naive blood monocytes, peritoneal macrophages and Kupffer cells as well as blood neutrophils.
- In the context of practical outcomes of research we will set experiments aimed to enhance efficacy of therapy with anthelmintic albendazole by means of co-administration of human immunological product with commercial name "Immodin“ (SevaPharma, CZ). Apart of study selected parameters of adaptive immune response we will focus on peritoneal exudate cells (macrophages, granulocytes and NK cells and cells isolated from liver granulomas (preliminary data are already available).

Technologies used

- Cells will be isolated from blood, peritoneal cavity and granulomatous inflammatory lesions in the liver of mice (following digestion with a set of enzymes).
- Established in vitro culture system of cells will be used for functional analyses to study suppressive activity towards lymphocytes and production of cytokines/chemokines, APC activity of macrophages
- Flow cytometry (BD FACSCantoTM , Becton Dickinson Biosciences) will be used for phenotypic studies, apoptosis, ROS production and phagocytosis.
- Molecular biology techniques: RT-PCR to examine gene expression profile in cells (for example: Fc receptors and MHC II, STAT6 and STAT3, genes for chemokines and cytokines)
- Immunohistochemistry for localization of selected markers on the liver sections and in isolated cells (light and fluorescent microscopy)
- In vitro culture system of larvae is available for collection of secretory products and study of interaction with cells (neutrophils, NK cells).
- ELISA system will be used for quantification of soluble markers (cytokines, chemokines, antibodies)
- SDS-PAGE, WB system and 2D electrophoresis for proteonomic studies and quantitative protein expression (for example, SOD, Bcl family of apoptotic proteins).
- Albendazole and Immodin are registered drugs for human administration.

 

Selected publications relevant to Mye-EUNITER

  1. HRČKOVÁ, Gabriela – VELEBNÝ, Samuel.  (2014) Application of Praziquantel in Experimental Therapy of Larval Cestodoses and Benefits of Combined Therapy and Drug Carriers. In:  QUICK W.  (ed)  Anthelmintics:  Clinical Pharmacology, Uses in Veterinary Medicine and Efficacy. NOVA Science Publisher, Inc.,  New York,  2014, p. 109-154.   ISBN 978-1-63117-714-9.      
  2. HRČKOVÁ, Gabriela – VELEBNÝ, Samuel. (2013) Pharmacological Potential of Selected Natural Compounds in the Control of Parasitic Diseases. Springer: Vienna, Heidelberg, New York, Dordrecht, p. 133.  ISSN 1864-8118,     ISBN 978-3-7091-1324-0.   DOI: 10.10007/978-3-7091-1325-7.          
  3. HRČKOVÁ, Gabriela – VELEBNÝ, Samuel – SOLÁR, P. (2010) Dynamics of hepatic stellate cells, collagen types I and III synthesis and gene expression of selected cytokines during hepatic fibrogenesis following Mesocestoides vogae (Cestoda) infection in mice. In: International Journal for Parasitology, vol. 40, no. 2, p. 163-174.  ISSN: 0020-7519
  4. HRČKOVÁ, Gabriela – VELEBNÝ, Samuel - DAXNEROVÁ, Zuzana - SOLÁR, Peter (2006) Praziquantel and liposomized-glucan treatment modulated liver fibrogenesis and mastocytosis in mice infected with Mesocestoides vogae (M.corti, Cestoda) tetrathyridia. In: Parasitology, vol. 132, no. 4, p. 581-594.  ISSN: 0031-1820 
  5. DVOROŽŇÁKOVÁ, Emília – HRČKOVÁ, Gabriela - BOROŠKOVÁ, Zora - VELEBNÝ, Samuel – DUBINSKÝ, Pavol (2004) Effect of treatment with free and liposomized albendazole on selected immunological parameters and cyst growth in mice infected with E. multilocullaris. In: Parasitology International,  vol. 53, no. 4, p. 315-325.  ISSN: 1383-5769

Contact details:

Institute of Parasitology of the Slovak Academy of Sciences
Hlinkova 3, 04001 Košice
Slovak Republic

Tel: 00421 556334455

hrcka@saske.sk