Highlights
- •IFT on HEp-2 cells is a valid alternative for the diagnosis of AIH when cut-off titers are increased.
- •ANA ELISA and F-actin ELISA represent potential alternatives to IFT for the diagnosis of AIH.
- •ANA ELISA kits should include HEp-2 nuclear extracts to account for unrecognized autoantigens.
- •ELISA cut-offs need to be validated locally to be predictive in diagnosing AIH.
Background & Aims
The simplified criteria for the diagnosis of autoimmune hepatitis (AIH) include immunofluorescence
testing (IFT) of antinuclear and smooth muscle autoantibodies (ANA and SMA) on rodent
tissue sections. We aimed to establish scoring criteria for the implementation of
ANA IFT on human epithelioma-2 (HEp-2) cells and ELISA-based testing.
Methods
ANA and SMA reactivity of 61 AIH sera and 72 non-alcoholic fatty liver disease controls
were separately assessed on tissue sections and HEp-2 cells to compare the diagnostic
value at increasing titers. A total of 113 patients with AIH at diagnosis and 202
controls from 3 European centers were assessed by IFT as well as 3 different commercially
available ANA ELISA and 1 anti-F-actin ELISA.
Results
ANA assessment by IFT on liver sections had 83.6% sensitivity and 69.4% specificity
for AIH at a titer of 1:40. On HEp-2 cells, sensitivity and specificity were 75.4%
and 73.6%, respectively, at an adjusted titer of 1:160. Area under the curve (AUC)
values of ANA ELISA ranged from 0.70–0.87, with ELISA coated with HEp-2 extracts in
addition to selected antigens performing significantly better. SMA assessment by IFT
had the highest specificity for the SMA-VG/T pattern and anti-microfilament reactivity
on HEp-2 cells. ELISA-based anti-F-actin evaluation was a strong predictor of AIH
(AUC 0.88) and performed better than SMA assessment by IFT (AUC 0.77–0.87).
Conclusion
At adjusted cut-offs, both ANA IFT using HEp-2 cells and ELISA-based autoantibody
evaluation for ANA and SMA are potential alternatives to tissue-based IFT for the
diagnosis of AIH.
Lay summary
Autoantibodies are a hallmark of autoimmune hepatitis and are traditionally tested
for by immunofluorescence assays on rodent tissue sections. Herein, we demonstrate
that human epithelioma cells can be used as a reliable substrate for immunofluorescence
testing. ELISA-based testing is also a potentially reliable alternative for autoantibody
assessment in autoimmune hepatitis. We propose the implementation of these testing
methods into the simplified criteria for the diagnosis of autoimmune hepatitis.
Graphical abstract
Graphical Abstract
Keywords
To read this article in full you will need to make a payment
Purchase one-time access:
Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online accessOne-time access price info
- For academic or personal research use, select 'Academic and Personal'
- For corporate R&D use, select 'Corporate R&D Professionals'
Subscribe:
Subscribe to Journal of HepatologyAlready a print subscriber? Claim online access
Already an online subscriber? Sign in
Register: Create an account
Institutional Access: Sign in to ScienceDirect
References
- Simplified criteria for the diagnosis of autoimmune hepatitis.Hepatology. 2008; 48: 169-176
- Liver autoimmune serology: a consensus statement from the committee for autoimmune serology of the International Autoimmune Hepatitis Group.J Hepatol. 2004; 41: 677-683
- Range of antinuclear antibodies in “healthy” individuals.Arthritis Rheum. 1997; 40: 1601-1611
- Challenges in the standardization of autoantibody testing: a comprehensive review.Clin Rev Allergy Immunol. 2017; 53: 68-77
- Sera with high levels of anti-smooth muscle and anti-mitochondrial antibodies frequently bind to cytoskeleton proteins.Clin Exp Immunol. 1990; 82: 52-56
- Classification of smooth muscle autoantibodies detected by immunofluorescence.J Clin Pathol. 1976; 29: 403-410
- Antibodies to filamentous actin (F-actin) in type 1 autoimmune hepatitis.J Clin Pathol. 2006; 59: 280-284
- Diagnostic relevance of anti-filamentous actin antibodies in autoimmune hepatitis.J Clin Pathol. 2007; 60: 107-108
- Smooth muscle antibodies and type 1 autoimmune hepatitis.Autoimmunity. 2002; 35: 497-500
- Patterns of nuclear immunofluorescence and reactivities to recombinant nuclear antigens in autoimmune hepatitis.Gastroenterology. 1994; 107: 200-207
- EASL Clinical Practice Guidelines: autoimmune hepatitis.J Hepatol. 2015; 63: 971-1004
- EASL Clinical Practice Guidelines: management of cholestatic liver diseases.J Hepatol. 2009; 51: 237-267
- EASL Clinical Practice Guidelines: the diagnosis and management of patients with primary biliary cholangitis.J Hepatol. 2017; 67: 145-172
- EASL-EASD-EASO Clinical Practice Guidelines for the management of non-alcoholic fatty liver disease.J Hepatol. 2016; 64: 1388-1402
- Immunomorphological characterisation of antinuclear antibodies in chronic liver disease.J Clin Pathol. 1985; 38: 801-805
- International recommendations for the assessment of autoantibodies to cellular antigens referred to as anti-nuclear antibodies.Ann Rheum Dis. 2014; 73: 17-23
- Enzyme-linked immunosorbent assay screening then indirect immunofluorescence confirmation of antinuclear antibodies: a statistical analysis.Am J Clin Pathol. 2011; 135: 678-684
- Comparison of an enzyme immunoassay to an indirect fluorescent immunoassay for the detection of antinuclear antibodies.Am J Clin Pathol. 1999; 111: 503-506
- Screening of antinuclear antibodies: comparison between enzyme immunoassay based on nuclear homogenates, purified or recombinant antigens and immunofluorescence assay.Clin Chem Lab Med. 2004; 42: 1155-1160
- Comparison of antinuclear antibody testing methods by ROC analysis with reference to disease diagnosis.Clin Chem. 1997; 43: 1987-1989
- Detection of antinuclear antibodies by solid-phase immunoassays and immunofluorescence analysis.Clin Chem. 2004; 50: 2141-2147
- Anti-ribosomal P protein: a novel antibody in autoimmune hepatitis.Liver Int. 2013; 33: 909-913
- Evaluation of F-actin ELISA for the diagnosis of autoimmune hepatitis.Am J Gastroenterol. 2006; 101: 2731-2736
Article info
Publication history
Published online: July 27, 2020
Accepted:
July 22,
2020
Received in revised form:
June 30,
2020
Received:
January 20,
2020
Identification
Copyright
© 2020 European Association for the Study of the Liver. Published by Elsevier B.V. All rights reserved.
