کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
1965702 1538680 2013 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
Development of a recombinant cell-based indirect immunofluorescence assay (RC-IFA) for the determination of autoantibodies against “rings and rods”-associated inosine-5′-monophosphate dehydrogenase 2 in viral hepatitis C
موضوعات مرتبط
علوم زیستی و بیوفناوری بیوشیمی، ژنتیک و زیست شناسی مولکولی زیست شیمی
پیش نمایش صفحه اول مقاله
Development of a recombinant cell-based indirect immunofluorescence assay (RC-IFA) for the determination of autoantibodies against “rings and rods”-associated inosine-5′-monophosphate dehydrogenase 2 in viral hepatitis C
چکیده انگلیسی

Background and aimsAutoantibodies against so-called “rings and rods” structures, as determined by indirect immunofluorescence assay (IFA) using the human cell line HEp-2, have been described in chronic hepatitis C virus (HCV) infected patients treated with interferon/ribavirin. Recently, cytidine triphosphate synthase (CTPS) and inosine-5′-monophosphate dehydrogenase 2 (IMPDH2), the enzyme inhibited by ribavirin, were proposed as the target antigens. We wanted to confirm the identification and setup a robust system for autoantibody testing in routine laboratories.MethodsCTPS and IMPDH2 were individually expressed in HEK293 cells and the recombinant cells were used in IFA (RC-IFA) to analyze sera from 33 anti-“rings and rods” antibody positive individuals with unknown diagnosis, 50 patients with chronic HCV infection, 100 with autoimmune hepatitis, 50 with primary biliary cirrhosis and 50 healthy blood donors.ResultsWe found that all sera with anti-“rings and rods” reacted with recombinant IMPDH2 but none with CTPS. In western blot or ELISA, anti-IMPDH2 positive sera reacted only weakly, if at all, with Escherichia coli derived recombinant IMPDH2 indicating that the autoantibody reaction probably depends on the 3-dimensional conformation of the antigen. A rabbit hyperimmune serum raised against bacterially expressed IMPDH2 produced the ring/rods pattern in IFA using HEp-2.ConclusionsWe conclude, that IMPDH2 is indeed the main target of anti-“rings and rods” while CTPS is an unlikely target. Moreover, the novel RC-IFA test system allows a standardized semi-quantitative determination of anti-IMPDH2 basing on a defined recombinant antigen.


► Confirmation of IMPDH2 as the main target of anti-rings/rods in HCV
► Development of a standardized test system
► Detection of antibodies in HCV samples that are negative by conventional IFA

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Clinica Chimica Acta - Volume 418, 15 March 2013, Pages 91–96
نویسندگان
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