Standardisation of the factor H autoantibody assay

Watson, Rochelle; Lindner, S.; Bordereau, Pauline; Hunze, Eva-Maria; Tak, F; Ngo, Stéphanie; Zipfel, Peter F.; Skerka, Christine; Dragon-Durey, Marie-Agnès; Marchbank, Kevin J.

doi:10.1016/j.imbio.2013.06.004

Cited by 49 publications

(39 citation statements)

References 29 publications

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“…The autoantibodies are detected by ELISA that uses purified factor H and allows determination of antibody titers and follow-up of titer changes in response to treatment. Our own experiments as well as a recently published comparison of various test methods support the originally described factor H autoantibody ELISA as a robust and well reproducible assay [59][60][61]. Some patients have IgA class autoantibodies in addition to IgG [62].…”

Section: Factor H Autoantibodiessupporting

confidence: 73%

Autoantibodies to complement components in C3 glomerulopathy and atypical hemolytic uremic syndrome

et al. 2014

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The alternative pathway of complement is implicated in the pathogenesis of several renal diseases, such as atypical hemolytic uremic syndrome, dense deposit disease and other forms of C3 glomerulopathy. The underlying complement defects include genetic and/or acquired factors, the latter in the form of autoantibodies. Because the autoimmune forms require a specific treatment, in part different from that of the genetic forms, it is important to detect the autoantibodies as soon as possible and understand their characteristics. In this overview, we summarize the types of anti-complement autoantibodies detected in such diseases, i.e. autoantibodies to factor H, factor I, C3b, factor B and those against the C3 convertases (C3 nephritic factor and C4 nephritic factor). We draw attention to newly described autoantibodies and their characteristics, and highlight similarities and differences in the autoimmune forms of these diseases.

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Section: Factor H Autoantibodiessupporting

confidence: 73%

Autoantibodies to complement components in C3 glomerulopathy and atypical hemolytic uremic syndrome

et al. 2014

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“…2b). Transformation of background subtracted OD450 value to RU using a titrated GB24 antibody as a standard curve (similar to (Watson et al, 2014)) confirmed no samples reached the nomimal 100 RU cut off for positivity used for determining positive titre in the FH autoantibody ELISA. However, this cut off maybe artificially high as comparing a mouse monoclonal to human serum has many technical pit falls.…”

Section: Resultsmentioning

confidence: 69%

“…Using the consensus ELISA protocol established in our recent FH autoantibody standardisation paper (Watson et al, 2014) as a template, we analysed 100 BDC samples and 89 aHUS patient samples for reactivity with the GB24 column purified Newcastle generatedrhCD46. We found relatively low reactivity of patient or control sera with coated plates ELISA (Fig.…”

Section: Resultsmentioning

confidence: 99%

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Autoantibodies to CD59, CD55, CD46 or CD35 are not associated with atypical haemolytic uraemic syndrome (aHUS)

Watson

Wearmouth

McLoughlin

et al. 2015

Molecular Immunology

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Autoantibody formation against Factor H (FH) is found in 7–10% of patients who are diagnosed with atypical haemolytic uraemic syndrome (aHUS). These autoantibodies predominately target the C-terminal cell binding recognition domain of FH and are associated with absence of FHR1. Additional autoantibodies have also been identified in association with aHUS, for example autoantibodies to Factor I. Based on this, and that there are genetic mutations in other complement regulators and activators associated with aHUS, we hypothesised that other complement regulator proteins, particularly surface bound regulators in the kidney, might be the target for autoantibody formation in aHUS. Therefore, we assayed serum derived from 89 patients in the Newcastle aHUS cohort for the presence of autoantibodies to CD46 (membrane cofactor protein, MCP), CD55 (decay accelerating factor, DAF), CD35 (complement receptor type 1, CR1; TP10) and CD59. We also assayed 100 healthy blood donors to establish the normal levels of reactivity towards these proteins in the general population. Recombinant proteins CD46 and CD55 (purified from Escherichia coli) as well as soluble CR1 (CD35) and oligomeric C4BP-CD59 (purified from eukaryotic cell media) were used in ELISA to detect high responders. False positive results were established though Western blot and flow cytometric analysis. After excluding false positive responders to bacterial proteins in the CD46 and CD55 preparations, and responses to blood group antigens in CD35, we found no significant level of patient serum IgG reactivity with CD46, CD55, CD35 or CD59 above that detected in the normal population. These results suggest that membrane anchored complement regulators are not a target for autoantibody generation in aHUS.

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“…The anti-CFH autoantibodies are generally measured by ELISA. Although several ELISA methods have been reported, Watson et al have recommended the use of the Paris method, which is the most robust, costeffective, and easy to establish [51].…”

Section: Complement Assessment In Ahusmentioning

confidence: 99%

Atypical hemolytic uremic syndrome

2017

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Atypical hemolytic uremic syndrome (aHUS) is a thrombotic microangiopathy (TMA) characterized by microangiopathic hemolytic anemia, thrombocytopenia, and acute renal failure. Most cases of aHUS are caused by uncontrolled complement activation due to genetic mutations in the alternative pathway of complement. More recently, mutations in the gene of coagulation system have also been identified in patients with aHUS. In Japan, the recent studies of aHUS have identified the unique genetic characteristics in our country and enabled us to revise the diagnostic criteria. In this article, we review the classification of TMAs and describe the pathophysiology, diagnosis, and management of aHUS. We also highlight current progress in clinical and basic research of the patients with aHUS in Japan.

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Standardisation of the factor H autoantibody assay

Cited by 49 publications

References 29 publications

Autoantibodies to complement components in C3 glomerulopathy and atypical hemolytic uremic syndrome

Autoantibodies to complement components in C3 glomerulopathy and atypical hemolytic uremic syndrome

Autoantibodies to CD59, CD55, CD46 or CD35 are not associated with atypical haemolytic uraemic syndrome (aHUS)

Atypical hemolytic uremic syndrome

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