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

Barwary, Nafal (2018) Use of the RS-ATL8 NFAT reporter system for diagnosis of hydatid disease. PhD thesis, University of Nottingham.

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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.

Item Type: Thesis (University of Nottingham only) (PhD)
Supervisors: Falcone, Franco
Winkler, Sebastiaan
Keywords: Parasitology, Immunology, Allergy, Medicine, Diagnosis, Molecular Biology
Subjects: Q Science > QL Zoology > QL360 Invertebrates
R Medicine > RA Public aspects of medicine > RA 421 Public health. Hygiene. Preventive Medicine
Faculties/Schools: UK Campuses > Faculty of Science > School of Pharmacy
Item ID: 55533
Depositing User: Barwary, Nafal
Date Deposited: 17 Dec 2018 15:26
Last Modified: 11 Dec 2020 04:30

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