Generation of monoclonal antibodies to murine bile duct epithelial cells: identification of annexin V as a new marker of small intrahepatic bile ducts.
Biliary epithelial cells (BECs) are distributed along the length of both the extrahepatic and intrahepatic biliary tree, but have distinctly different phenotypes and functions according to their anatomical location. It has been reasoned that the distinct appearance of pathogenic lesions in different biliary diseases may be associated with the expression of distinct proteins. These data prompted us to immunize rats with cultured murine BECs with the objective of determining if there are unique antigens on BECs. Of the 45 monoclonal antibodies (mAbs) produced, 12 mAbs (MBEC 1-12) were selected for detailed study based on their classification into three major groups. These groups included four antibodies (MBEC 1-4) that reacted in a staining pattern typical of mucin. A second group of mAbs, MBECs 5 to 8, reacted strongly along the biliary tract and by immunoblot analysis, reacted with several bands ranging from 44 kd to 64 kd. These antibodies were considered as markers of pan BECs and their staining pattern proved similar to that of a control polyclonal pan-cytokeratin. The final group of mAbs, MBECs 9 to 12, recognized a 36-kd antigen using lysates of murine BECs. These antibodies also predominantly stained small peripheral bile ducts. The reactive antigen was purified by immunoprecipitation and microsequenced; the peptides sequenced showed 100% homology with murine annexin V. The identification of annexin V with predominantly intrahepatic bile ducts, is of significant interest because of the multiple roles of annexin V, including that of membrane cytoskeletal interactions during transport and apoptosis.
Katayanagi
K
K
Division of Rheumatology, Allergy and Clinical Immunology, University of California at Davis, School of Medicine, CA, USA.
Van de Water
J
J
Kenny
T
T
Nakanuma
Y
Y
Ansari
A A
AA
Coppel
R
R
Gershwin
M E
ME
D000595:Amino Acid Sequence; D000818:Animals; D017304:Annexin A5 / Q000276:immunology / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000096:biosynthesis*; D000918:Antibody Specificity; D001653:Bile Ducts, Intrahepatic / Q000378:metabolism*; D015415:Biomarkers / Q000032:analysis; D002478:Cells, Cultured; D004847:Epithelial Cells / Q000276:immunology / Q000378:metabolism; D005260:Female; D006825:Hybridomas / Q000276:immunology / Q000378:metabolism; D007150:Immunohistochemistry; D051379:Mice; D008807:Mice, Inbred BALB C; D008969:Molecular Sequence Data; D008970:Molecular Weight; D009928:Organ Specificity; D011233:Precipitin Tests; D051381:Rats; D017421:Sequence Analysis
D017304:Annexin A5; D000911:Antibodies, Monoclonal; D015415:Biomarkers