Title : ( The effects of germinal layer antigens of hydatid cyst on the expression level of ovine Toll-like Receptor 2 and 4( TLR2 and TLR4) in Peripheral blood mononuclear cells (PBMCs) )
Authors: fatemeh taran , nooshinmehr soleymani , Hassan Borji , M. Torabi , M.H. Nazemshirazi , Mohammad Azizzadeh , Alireza Haghparast ,Access to full-text not allowed by authors
Abstract
Background: Echinococcosis is one of the most important zoonosis disease throughout the world and represents a major public health and economic burden in many countries. Echinococcosis Granolosus causesd hydatid cyst in human and domesticated animals. Hydatid cyst develops in many organs, mostly in liver and lungs. Immune responses against hydatidosis comprises of various mechanisms of innate and adaptive immunity. In recent years the importance of innate immune responses and in particular, pattern recognition receptors (PRRs) has been recognized as an essential mechanisms for development of an effective immune response. PRRs are the main sensors of pathogen and danger signals in innate immunity. Toll like receptors (TLRs) are the most studied and best characterized PPRs which are responsible for sensing pathogen associated molecular patterns (PAMPs). The role of TLRs in the molecular mechanisms underlying the pathogenesis and immunity in helminthes infection has not been clearly defined. Methods: In this study we focused on the expression levels of two important ovine TLRs transcripts, namely TLR2 and TLR4 in a culture of ovine lymphocytes exposed to different concentrations of germinal layer antigens of hydatid cysts in a time point experiments. Blood samples were taken from healthy young lambs. After isolation of peripheral blood mononuclear cells (PBMC), the cells were cultured with different concentrations(50µg/ml &100µg/ml) of germinal layer antigens (antigens were extracted and concentrated according to the standard protocols) in different time points. Then, total RNA was isolated from the cell pellets and cDNA was synthesized using Oligo dT primers. ThenAfterwards, the primer pairs for TLR2, TLR4 as target genes and GAPDH as housekeeping and calibrator gene were optimized in a gradient RT-PCR experiment. Subsequently, qPCR analysis was set up to quantify and compare the relative expression levels of TLR2 and TLR4 transcripts in PBMC of antigen treated samples versus control (untreated) samples. Results: Statistical analysis using one way T-test showed an up-regulation of TLR2 and TLR4 in treated as compared to the untreated control group. Moreover, the increased expression of TLR4 was significant. Conclusion: the results presented in this study, can shed more lights to the insight mechanisms behind the molecular immuopathogenesis of hydatidosis which might eventually pave the way for designing novel and more effective therapeutic as well as preventive strategies.
Keywords
, Echinococcosis Granolosus, hydatidosis, Gene expression, Innate Immunity, Toll Like Receptors(TLRs), real time quatitative PCR (qPCR)@inproceedings{paperid:1029047,
author = {Taran, Fatemeh and Soleymani, Nooshinmehr and Borji, Hassan and M. Torabi and M.H. Nazemshirazi and Azizzadeh, Mohammad and Haghparast, Alireza},
title = {The effects of germinal layer antigens of hydatid cyst on the expression level of ovine Toll-like Receptor 2 and 4( TLR2 and TLR4) in Peripheral blood mononuclear cells (PBMCs)},
booktitle = {11th international Congress of Immunology & Allergy},
year = {2012},
location = {تهران, IRAN},
keywords = {Echinococcosis Granolosus; hydatidosis; Gene expression; Innate Immunity; Toll Like Receptors(TLRs); real time quatitative PCR (qPCR)},
}
%0 Conference Proceedings
%T The effects of germinal layer antigens of hydatid cyst on the expression level of ovine Toll-like Receptor 2 and 4( TLR2 and TLR4) in Peripheral blood mononuclear cells (PBMCs)
%A Taran, Fatemeh
%A Soleymani, Nooshinmehr
%A Borji, Hassan
%A M. Torabi
%A M.H. Nazemshirazi
%A Azizzadeh, Mohammad
%A Haghparast, Alireza
%J 11th international Congress of Immunology & Allergy
%D 2012