Volume 39, Issue 5, Pages (November 2003)

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Volume 39, Issue 5, Pages 800-806 (November 2003) Cytochrome P450 2A6: a new hepatic autoantigen in patients with chronic hepatitis C virus infection Georgios N Dalekos, Petra Obermayer-Straub, Moritz Bartels, Takashi Maeda, Anne Kayser, Sabine Braun, Stephanie Loges, Eleonore Schmidt, M.Eric Gershwin, Michael P Manns Journal of Hepatology Volume 39, Issue 5, Pages 800-806 (November 2003) DOI: 10.1016/S0168-8278(03)00356-8

Fig. 1 Detection of anti-CYP2A6 (A) and anti-CYP1A2 (B) by the radioligand assay in several groups of patients. CYP2A6, cytochrome P450 2A6; CYP1A2, cytochrome P450 1A2; HCV/LKM-1+ and −, hepatitis C virus/positive or negative for liver kidney microsomal antibodies type 1, respectively; HBV, hepatitis B virus infection; HDV, hepatitis D virus; HGV, hepatitis G/GBVC virus; AIH-1, autoimmune hepatitis type 1; AIH-2, autoimmune hepatitis type 2; PBC, primary biliary cirrhosis; PSC, primary sclerosing cholangiitis; ARD, autoimmune rheumatic diseases; HC, healthy controls. Every sample with values more than the mean value of healthy controls plus 4 S.D.s was considered positive (cut-off point of the assays: CYP2A6: 650 cpm; CYP1A2 700 cpm). Journal of Hepatology 2003 39, 800-806DOI: (10.1016/S0168-8278(03)00356-8)

Fig. 2 Results of immunoprecipitation experiments with [35S]-CYP2A6 (A) and [35S]-CYP2D6 (B) as target antigens. [35S]-CYP2A6 and [35S]-CYP2D6 were visualized after SDS-polyacrylamide gel electrophoresis (SDS-PAGE) by means of a Fujix Bas 1000 bioimaging analyzer. Lane 1: signal of 30.000 cpm [35S]-CYP2A6 and [35S]-CYP2D6 proteins, respectively; lanes 2–5: HCV sera positive for both anti-CYP2A6 and anti-CYP2D6 autoantibodies; lanes 6–8: anti-CYP2A6 positive sera from patients with HCV, HGV/GBVC, AIH-1, respectively, lane 9: serum from a healthy blood donor; lane 10: serum from a patient with autoimmune polyglandular syndrome type 1 (APS-1) strongly positive for anti-CYP2A6 only; lanes 11, 12: AIH-2 serum and HCV serum, respectively, positive for anti-CYP2D6; lane 13: HCV serum negative for both anti-CYP2A6 and anti-CYP2D6 autoantibodies tested; lane 14: HGV/GBVC serum negative for both anti-CYP2A6 and anti-CYP2D6 autoantibodies tested; abbreviations are same as in the text. Journal of Hepatology 2003 39, 800-806DOI: (10.1016/S0168-8278(03)00356-8)

Fig. 3 Staining pattern of monospecific anti-CYP2A6 autoantibodies on cryosections of rat liver (A) and rat kidney (B). Cytoplasmic staining of the whole liver lobule (A) and staining of the proximal renal tubules (B) in a serum with high titers of anti-CYP2A6 antibodies from a patient with autoimmune polyglandular syndrome type 1 (APS-1). This pattern cannot be distinguished from the anti-LKM-1 staining on the basis of IIF. Expression of other antibodies against several cytochromes P450 was excluded by tests on Western blots with a panel of 10 different recombinant hepatic cyto-chromes P450 (CYP2D6, CYP1A1, CYP1A2, CYP2A6, CYP3A4, CYP2B6, CYP2C8, CYP2C9, CYP2C19 and CYP2E1). Only anti-CYP2A6 antibodies were detectable on these blots. Tests by the anti-LKM-1 ELISA, and the anti-CYP2D6 and anti-CYP1A2 radioligand assays were also negative. Journal of Hepatology 2003 39, 800-806DOI: (10.1016/S0168-8278(03)00356-8)