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Chapter 122 : Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis

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Abstract:

This chapter focuses on the detection of antimitochondrial autoantibodies (AMA) in primary biliary cirrhosis (PBC) and the detection of liver-kidney microsomal antibodies (LKM antibodies) in autoimmune hepatitis (AIH). PBC is an autoimmune disease of the liver characterized by autoimmunity-mediated destruction of intrahepatic bile ducts with progressive inflammatory scarring and, eventually, liver function failure. Immunologically, PBC is characterized by the presence of AMA in the circulation and the infiltration of T cells into the liver. AMA can be detected by the following methods: (i) indirect immunofluorescence (IF), (ii) immunoblotting, and (iii) enzyme-linked immunosorbent assays (ELISAs). LKM antibodies have been defined due to their typical immunofluorescent staining of liver and kidney tissue. LKM autoantibodies can be detected by the following methods: (i) IF, (ii) competitive ELISA, (iii) immunoblotting with human liver microsomes, and (iv) immunoblotting with recombinant antigens.

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122

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Figures

Image of FIGURE 1
FIGURE 1

Detection of AMA by IF on HEp-2 cells. Note the characteristic mitochondrial staining pattern by PBC sera.

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
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Image of FIGURE 2
FIGURE 2

Reactivity of PBC sera to bovine heart mitochondria. Bovine heart mitochondria were resolved by SDS-PAGE, transferred to a nitrocellulose filter, and tested for their reactivity with PBC sera (lanes 1 to 5). Note the different patterns of AMA reactivities for various patients with PBC to the 70-kDa PDC-E2, 55-kDa E3 binding protein (protein X), 52-kDa BCOADC-E2, 48-kDa OGDC-E2, and 41-kDa PDC-E1a proteins. Sera from healthy controls do not react to any of these mitochondrial proteins (lane 6).

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
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Image of FIGURE 3
FIGURE 3

Detection of LKM antibodies and AMA by IF on cryostat rat liver and kidney sections. (A) Staining of liver lobule by LKM-1 antibodies. (B) Staining of proximal but not distal renal tubules by LKM-1 antibodies.

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
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References

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Tables

Generic image for table
TABLE 1

Summary of 2-oxo-acid dehydrogenase mitochondrial antigens in PBC

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
Generic image for table
TABLE 2

Clinical characteristics of PBC

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
Generic image for table
TABLE 3

Diagnostic criteria for AIH

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122
Generic image for table
TABLE 4

Heterogeneity of microsomal antigens

Citation: Leung P, Manns M, Coppel R, Gershwin M. 2006. Detection of Antimitochondrial Autoantibodies in Primary Biliary Cirrhosis and Liver Kidney Microsomal Antibodies in Autoimmune Hepatitis, p 1082-1090. In Detrick B, Hamilton R, Folds J (ed), Manual of Molecular and Clinical Laboratory Immunology, 7th Edition. ASM Press, Washington, DC. doi: 10.1128/9781555815905.ch122

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