Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease

Despite major advances in the diagnosis of infections for many pathogenic organisms, there is still a problem obtaining an accurate immunodiagnosis of Echinococcus infection due to its serological cross-reactivity with other species of taeniid cestodes or at higher taxonomic levels. For this reason,...

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Main Author: Barwary, Nafal
Format: Thesis (University of Nottingham only)
Language:English
Published: 2018
Subjects:
Online Access:https://eprints.nottingham.ac.uk/55533/
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author Barwary, Nafal
author_facet Barwary, Nafal
author_sort Barwary, Nafal
building Nottingham Research Data Repository
collection Online Access
description Despite major advances in the diagnosis of infections for many pathogenic organisms, there is still a problem obtaining an accurate immunodiagnosis of Echinococcus infection due to its serological cross-reactivity with other species of taeniid cestodes or at higher taxonomic levels. For this reason, there are ongoing efforts to develop a better method for diagnosis of echinococcosis, especially when the parasite has a crucial role in hypersensitivity reactions. Like other helminthic infections, one of the immunological hallmarks is an elevated serum concentration of parasite-specific IgE. Our aim was to assess the use of IgE reporter system as a possible new method for diagnosis Echinococcus spp infection, using RS-ATL8 NFAT Reporter System, which is a humanised rat basophilic leukaemia (RBL) cell line that can be used to detect the presence of specific human IgE directed against Echinococcus allergens and cross-link their receptors, depending on luciferase generation as an indication of presence of parasite-specific IgE, pointing to infection. Towards this goal, we first optimised the use of the humanised RS-ATL8 Reporter System. This was achieved by optimisation of experimental conditions, such as cell density, stimulation time, optimum conditions for sensitising factors and stimulant optimum concentration. Once a robust standard operating procedure had been elaborated, the second goal was to choose a few Echinococcus antigens for the investigation into their immunogenic properties and potential diagnostic value and to express them recombinantly for testing through the RS-ATL8 NFAT Reporter System. The chosen antigens were EF1-alpha, EgAg5, AgB2, Cyclophilin, Eg19, EgTeg, and EgTPx.
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spelling nottingham-555332025-02-28T14:18:05Z https://eprints.nottingham.ac.uk/55533/ Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease Barwary, Nafal Despite major advances in the diagnosis of infections for many pathogenic organisms, there is still a problem obtaining an accurate immunodiagnosis of Echinococcus infection due to its serological cross-reactivity with other species of taeniid cestodes or at higher taxonomic levels. For this reason, there are ongoing efforts to develop a better method for diagnosis of echinococcosis, especially when the parasite has a crucial role in hypersensitivity reactions. Like other helminthic infections, one of the immunological hallmarks is an elevated serum concentration of parasite-specific IgE. Our aim was to assess the use of IgE reporter system as a possible new method for diagnosis Echinococcus spp infection, using RS-ATL8 NFAT Reporter System, which is a humanised rat basophilic leukaemia (RBL) cell line that can be used to detect the presence of specific human IgE directed against Echinococcus allergens and cross-link their receptors, depending on luciferase generation as an indication of presence of parasite-specific IgE, pointing to infection. Towards this goal, we first optimised the use of the humanised RS-ATL8 Reporter System. This was achieved by optimisation of experimental conditions, such as cell density, stimulation time, optimum conditions for sensitising factors and stimulant optimum concentration. Once a robust standard operating procedure had been elaborated, the second goal was to choose a few Echinococcus antigens for the investigation into their immunogenic properties and potential diagnostic value and to express them recombinantly for testing through the RS-ATL8 NFAT Reporter System. The chosen antigens were EF1-alpha, EgAg5, AgB2, Cyclophilin, Eg19, EgTeg, and EgTPx. 2018-12-11 Thesis (University of Nottingham only) NonPeerReviewed application/pdf en arr https://eprints.nottingham.ac.uk/55533/1/Thesis%20Final%20Version%2011-11-2018.pdf Barwary, Nafal (2018) Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease. PhD thesis, University of Nottingham. Parasitology Immunology Allergy Medicine Diagnosis Molecular Biology
spellingShingle Parasitology
Immunology
Allergy
Medicine
Diagnosis
Molecular Biology
Barwary, Nafal
Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title_full Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title_fullStr Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title_full_unstemmed Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title_short Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease
title_sort use of the rs-atl8 nfat reporter system for diagnosis of hydatid disease
topic Parasitology
Immunology
Allergy
Medicine
Diagnosis
Molecular Biology
url https://eprints.nottingham.ac.uk/55533/