Immunoblotting as a confirmatory test for antimitochondrial antibodies in primary biliary cirrhosis.

Provenzano, G.; Diquattro, Orazia; Craxı̀, Antonio; Almasio, Piero Luigi; Pinzello, Giovambattista; Marino, L; Fiorentino, Gabriella; Rinaldi, F; Pagliaro, L

doi:10.1136/gut.34.4.544

Cited by 13 publications

(4 citation statements)

References 21 publications

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“…We found that the frequency of serum AMA in patients with AIH was 34%, relatively higher than in previous reports. In AIH patients, AMA have been sought by IIF [13][14][15][16][17] and ELISA. [16][17][18] These data demonstrate that the prevalence of AMA in AIH sera is 5-10% by IIF and ELISA.…”

Section: Discussionmentioning

confidence: 99%

Presence of antimitochondrial autoantibodies in patients with autoimmune hepatitis

Nezu¹,

Tanaka

Yasui

et al. 2006

J of Gastro and Hepatol

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Section: Discussionmentioning

confidence: 99%

Presence of antimitochondrial autoantibodies in patients with autoimmune hepatitis

Nezu¹,

Tanaka

Yasui

et al. 2006

J of Gastro and Hepatol

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“…However, AMA cannot be detected in serum from about 10% of patients with clinical and pathological features of PBC. 7 In a previous study, we reported that anti-M2 antibody has four major proteins (70, 50, 47, 40 kDa) and was detected in both AMA-positive and AMA-negative PBC patients by the immunoblotting method with IgG, IgA, and IgM subclasses. 8,9 Recently, AMA-negative PBC in which AMA and anti-M2 antibodies are not detected but a high titer of antinuclear antibody (ANA) is present in the serum was proposed as autoimmune cholangitis (AIC).…”

Section: Introductionmentioning

confidence: 98%

Detection of IgA, IgM, and IgG subclasses of anti-M2 antibody by immunoblotting in autoimmune cholangitis: Is autoimmune cholangitis an early stage of primary biliary cirrhosis?

Nakajima

Shimizu

Miyazaki

et al. 1999

Journal of Gastroenterology

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Autoimmune cholangitis (AIC) has been proposed as a distinct disease entity from primary biliary cirrhosis (PBC), without antimitochondrial antibody (AMA) and anti-M2 antibody but with a high titer of antinuclear antibody (ANA) in the serum. However, negativity for AMA and anti-M2 antibody was determined by different methods in different studies. We hypothesized that anti-M2 antibody negativity in AIC resulted from methodological differences, including selection of the immunoglobulin subclass of the autoantibody. Twenty-three patients compatible with AIC whose serum tested negative for AMA and positive for ANA (> or = 1:80) were compared with 71 AMA-positive PBC patients. Laboratory findings, histology, and the pattern of anti-M2 antibody assessed by immunoblotting were compared. Alkaline phosphatase, total bilirubin, total cholesterol, and IgM values were lower in patients with AIC (P < 0.05, 0.01, respectively). Anti-smooth muscle antibody was detected more frequently in patients with AIC (P < 0.01). However, anti-M2 antibody was detected using immunoblotting not only in PBC but also in AIC cases. IgA class alone, IgM class alone, or both IgA and IgM classes of anti-M2 antibody were detected in 13%, 17%, and 22% of AIC patients, respectively, whereas they were not detected in PBC patients (P < 0.05, P < 0.01, P < 0.01). IgG class anti-M2 was detected in all patients with PBC, whereas it was detected in 48% of patients with AIC (P < 0.01). Histological evaluation showed that the early stages of disease were found more frequently in AIC (78%) than in PBC patients (39%) (P < 0.01). Anti-M2 antibody was detected by immunoblotting in all AIC patients. Hence, AIC is not a distinct disease from PBC. For diagnosing AIC and/or PBC, anti-M2 antibody should be examined by the immunoblotting assay to detect not only IgG but also IgA and IgM subclasses.

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“…However, it may be difficult to discern AMA from concurrent antibodies e.g. liver/kidney microsomal antibodies when using HEp-2 cells [69]. With the multiorgan substrate, AMA will preferentially stain distal over proximal renal tubules as well as the gastric cells.…”

Section: Detection Of Amamentioning

confidence: 99%

“…For example, in a study performed by Provenzano, et al, 16 non-PBC patients had positive AMA by IIF but only 3 were confirmed to be AMA positive by immunoblotting. In addition, over the course of two years, 18 out of 34 patients with positive AMA by IIF were subsequently diagnosed as PBC [69]. In spite of such limitations, IIF is still a sufficiently sensitive method that will be useful in diagnosis when utilized with other assays [71].…”

Section: Detection Of Amamentioning

confidence: 99%

A contemporary perspective on the molecular characteristics of mitochondrial autoantigens and diagnosis in primary biliary cholangitis

Leung

Choi

Yang

et al. 2016

Expert Review of Molecular Diagnostics

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Primary biliary cholangitis (PBC) is an autoimmune hepatobiliary disease characterized by immune mediated destruction of the intrahepatic small bile ducts and the presence of antimitochondrial antibodies (AMAs). The mitochondrial autoantigens have been identified as the E2 subunits of the 2-oxo-acid dehydrogenase complex, including the E2 subunits of pyruvate dehydrogenase, branched-chain 2-oxo acid dehydrogenase complex, oxoglutarate dehydrogenase complex, E3 binding protein and PDC E1 alpha subunit. The AMA epitope is mapped within the E2 lipoic acid binding domain, which is particularly important for oxidative phosphorylation. In addition, lipoic acid, which serves as a swinging arm to capture electrons, is particularly susceptible to an electrophilic attack and may provide clues to the etiology of PBC. This review emphasizes the molecular characteristics of AMAs, including detection, immunochemistry and the putative role in disease. These data have significance not only specifically for PBC, but generically for autoimmunity.

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Immunoblotting as a confirmatory test for antimitochondrial antibodies in primary biliary cirrhosis.

Cited by 13 publications

References 21 publications

Presence of antimitochondrial autoantibodies in patients with autoimmune hepatitis

Presence of antimitochondrial autoantibodies in patients with autoimmune hepatitis

Detection of IgA, IgM, and IgG subclasses of anti-M2 antibody by immunoblotting in autoimmune cholangitis: Is autoimmune cholangitis an early stage of primary biliary cirrhosis?

A contemporary perspective on the molecular characteristics of mitochondrial autoantigens and diagnosis in primary biliary cholangitis

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