Formiminotransferase cyclodeaminase is an organ-specific autoantigen recognized by sera of patients with autoimmune hepatitis Pascal Lapierre, Oumnia.

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Formiminotransferase cyclodeaminase is an organ-specific autoantigen recognized by sera of patients with autoimmune hepatitis Pascal Lapierre, Oumnia Hajoui, Jean-Claude Homberg, Fernando Alvarez Gastroenterology Volume 116, Issue 3, Pages 643-649 (March 1999) DOI: 10.1016/S0016-5085(99)70186-1 Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 1 Human hepatoma HepG2 cell line expresses the LC1 antigen. (A) A 62-kilodalton (kDa) protein in 10% SDS-PAGE is immunoprecipitated by anti-LC1– and LKM1/LC1–positive sera from [35S]cysteine-labeled HepG2 proteins. The 48-kilodalton antigen specifically recognized by LKM1-positive sera is not present in the HepG2 cell line. (B) Northern blot analysis shows that the LC1 antigen (FTCD) mRNA is present in human liver and HepG2 cells, but the LKM1 antigen (P450 2D6) is not found in HepG2 cells, confirming that the HepG2 cell line is a good model for identification of the LC1 antigen. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 1 Human hepatoma HepG2 cell line expresses the LC1 antigen. (A) A 62-kilodalton (kDa) protein in 10% SDS-PAGE is immunoprecipitated by anti-LC1– and LKM1/LC1–positive sera from [35S]cysteine-labeled HepG2 proteins. The 48-kilodalton antigen specifically recognized by LKM1-positive sera is not present in the HepG2 cell line. (B) Northern blot analysis shows that the LC1 antigen (FTCD) mRNA is present in human liver and HepG2 cells, but the LKM1 antigen (P450 2D6) is not found in HepG2 cells, confirming that the HepG2 cell line is a good model for identification of the LC1 antigen. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 2 Similarities between the FTCD pig sequence and the LCHC1 sequence: (A) nucleotides and (B) amino acid sequences. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 3 FTCD is the liver-specific antigen recognized by anti-LC1s. (A) LCHC1 recombinant fusion protein test by immunoblot, containing the C-terminal region of human FTCD, is recognized only by LC1/LKM1- and anti-LC1–positive sera. In total, 20 anti-LC1–positive sera (alone or in association with LKM1s) recognize the LCHC1 recombinant fusion protein. (B) Cross-reactivity between LCHC1 recombinant fusion protein and the human liver cytosol 62-kilodalton protein. The anti-FTCD affinity-purified antibodies (lane b) and the sera from mice immunized with the LCHC1 recombinant fusion protein react against a human liver 62-kilodalton cytosolic protein in an immunoblot assay. Lane a, anti-LC1–positive serum; lane c, anti-LC1–negative serum; lanes d–f, mouse sera. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 3 FTCD is the liver-specific antigen recognized by anti-LC1s. (A) LCHC1 recombinant fusion protein test by immunoblot, containing the C-terminal region of human FTCD, is recognized only by LC1/LKM1- and anti-LC1–positive sera. In total, 20 anti-LC1–positive sera (alone or in association with LKM1s) recognize the LCHC1 recombinant fusion protein. (B) Cross-reactivity between LCHC1 recombinant fusion protein and the human liver cytosol 62-kilodalton protein. The anti-FTCD affinity-purified antibodies (lane b) and the sera from mice immunized with the LCHC1 recombinant fusion protein react against a human liver 62-kilodalton cytosolic protein in an immunoblot assay. Lane a, anti-LC1–positive serum; lane c, anti-LC1–negative serum; lanes d–f, mouse sera. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions

Fig. 4 Ouchterlony double immunodiffusion method showing precipitation lines of identity between anti-LC1s and anti-pig FTCD polyclonal antibodies. Cytosol, liver cytosol subcellular fraction (2.5 mg); LC1, liver cytosol type 1–positive serum from a patient with AIH; anti-pig FTCD, rabbit polyclonal anti-pig FTCD antibody (a gift from Dr. R. E. MacKenzie, McGill University, Montréal, Québec, Canada) that cross-reacts with human FTCD. Gastroenterology 1999 116, 643-649DOI: (10.1016/S0016-5085(99)70186-1) Copyright © 1999 American Gastroenterological Association Terms and Conditions