Autoimmune liver disease-related autoantibodies in patients with biliary atresia

Table 1 Demographic and clinical features, and biochemical parameters¹ of biliary atresia patients and non-biliary atresia controls n (%)

Variable	BA, n = 124	Non-BA, n = 140
		Other liver diseases2, n = 92	Healthy, n = 48
Age, mo	2.9 (1.9-3.0)	2.8 (2.0-3.2)	3.4 (2.0-4.0)
Sex, male/female	60/64	50/42	25/23
METAVIR score
F0	4 (3.2)	NA	NA
F1	37 (29.8)	NA	NA
F2	24 (19.4)	NA	NA
F3	50 (40.3)	NA	NA
F4	9 (7.3)	NA	NA
ALT, U/L	162.4 (104.3-204.6)a	132.0 (70.5-247.5)	28.1 (22.4-40.8)
AST, U/L	190.5 (151.4-257.4)a	195.9 (117.0-292.5)	38.0 (26.0-52.3)
γ-GT, U/L	716.0 (411.0-1142.5)a	598.4 (189.5-1043.2)	32.7 (23.4-46.3)
ALP, U/L	406.8 (316.5-522.2)a	517.0 (409.7-660.3)	275.8 (189.0-345.6)
TBIL, μmol/L	157.0 (126.5-184.0)a	145.7 (107.1-196.5)	6.5 (2.8-14.8)
DBIL, μmol/L	130.5 (103.5-152.7)a	110.1 (88.4-137.8)	3.1 (1.0-4.9)

Data are described as median (interquartile range: 25^th-75^th percentile). ¹Reference intervals: ALT, 3-35 U/L; AST, 5-60 U/L; γ-GT, 13-57 U/L; ALP, 118-390 U/L; TBIL, 2-17 μmol/L; DBIL, 0-7 μmol/L;

²Choledochal cysts, transient cholestasis of unknown origin, and neonatal intrahepatic cholestasis caused by citrin deficiency were included as disease controls.

^aP < 0.05 vs healthy controls. ALP: Alkaline phosphatase; ALT: Alanine aminotransferase; AST: Aspartate aminotransferase; BA: Biliary atresia; DBIL: Direct bilirubin; FO-F4: Fibrosis scores 0-4; γ-GT: Gamma-glutamyl transpeptidase; NA: Not applicable; TBIL: Total bilirubin.

Table 2 Prevalence profile of autoimmune liver disease in biliary atresia patients and non-biliary atresia controls n (%)

Variable	BA, n = 81	Non-BA, n = 88
		Other liver diseases1, n = 44	Healthy, n = 40	Total
PBC-related antibodies	15 (18.5)c	4 (9.1)	1 (2.5)	5 (5.7)
AMA-M2	1 (1.2)	0 (0)	0 (0)	0 (0)
Anti- BPO	12 (14.8)ac	2 (4.5)	0 (0)	2 (2.2)
Anti-Sp100	1 (1.2)	0 (0)	0 (0)	0 (0)
Anti-gp210	2 (2.5)	1 (2.3)	1 (2.5)	2 (2.2)
Anti-PML	3 (3.7)	1 (2.3)	0 (0)	1 (1.1)
AMA-M2 + anti-BPO	1 (1.2)	0 (0)	0 (0)	0 (0)
AMA-M2 + anti-BPO + anti-Sp100 + anti-PML	1 (1.2)	0 (0)	0 (0)	0 (0)
AIH-related antibodies	6 (7.4)	3 (6.8)	3 (7.5)	6 (6.8)
Anti-LKM-1	0 (0)	0 (0)	0 (0)	0 (0)
Anti-LC-1	5 (6.2)	3 (6.8)	3 (7.5)	6 (6.8)
Anti-SLA/LP	1 (1.2)	0 (0)	0 (0)	0 (0)
Anti-Ro-52	5 (6.2)	2 (4.5)	2 (5)	4 (4.5)

¹Choledochal cysts, transient cholestasis of unknown origin, and neonatal intrahepatic cholestasis caused by citrin deficiency were included as disease controls. AMA-M2 + M2-3E: combined the positivity to AMA-M2 and BPO; AMA-M2 + BPO + Sp100 + PML: Combined the positivity to AMA-M2, BPO, Sp100, and PML.

^cP < 0.05 vs non-BA;

^aP < 0.05 vs healthy controls. AIH: Autoimmune hepatitis; BA: Biliary atresia; PBC: Primary biliary cholangitis; LKM-1: Liver-kidney microsomal type 1; LC-1: Liver cytosolic antigen type 1.

Table 3 Prevalence of anti-nuclear antibodies in biliary atresia patients and non-biliary atresia controls n (%)

Variable	BA, n = 124	Other liver diseases1, n = 92	Healthy, n = 48
ANA, by IIF	14 (11.3)c	3 (3.3)	2 (4.2)
Fluorescence patterns
Homogeneous	3 (2.4)	0 (0)	0 (0)
Speckled	3 (2.4)	0 (0)	1 (2.1)
Nucleolar	3 (2.4)	0 (0)	1 (2.1)
Nuclear dots	1 (0.8)	0 (0)	0 (0)
Centrosome	1 (0.8)	0 (0)	0 (0)
Cytoplasm	1 (0.8)	0 (0)	0 (0)
Ring or rod Golgi	1 (0.8)	2 (2.2)	0 (0)
Centromere	1 (0.8)	0 (0)	0 (0)
Spindle apparatus	0 (0)	1 (1.1)	0 (0)
Specific ANA2, by line-blot
SSA	1 (0.8)	0 (0)	0 (0)
Ro-52	5 (4.0)	2 (2.2)	2 (4.2)
CENP B	4 (3.2)	1 (1.1)	0 (0)
dsDNA	3 (2.4)	0 (0)	0 (0)

¹Choledochal cysts, transient cholestasis of unknown origin, and neonatal intrahepatic cholestasis caused by citrin deficiency were included as disease controls;

²Specific ANAs included 12 different antibodies, with only SSA, Ro-52, CENP B, and dsDNA positive in the study.

^cP < 0.05 vs other liver diseases. ANA: Anti-nuclear antibody; BA: Biliary atresia; IIF: Indirect immunofluorescence.

Table 4 Prevalence of anti-neutrophil cytoplasmic antibodies in biliary atresia patients and non-biliary atresia controls n (%)

Variable	BA, n = 124	Other liver diseases1, n = 92	Healthy, n = 48
ANCA, by IIF	36 (29.0)a	23 (25.0)	2 (4.2)
c-ANCA	10 (8.1)	1 (1.1)	0 (0)
p-ANCA	25 (20.2)	21 (22.8)	2 (4.2)
a-ANCA	1 (0.8)	1 (1.1)	0 (0)
Specific ANCA, by ELISA
Anti-MPO	11 (8.9)	6 (6.5)	2 (4.2)
Anti-PR3	19 (15.3)	13 (14.2)	0 (0)
Anti-MPO and/or anti-PR3	23 (18.0)a	13 (14.2)	2 (4.2)
ANCAs	47 (37.9)a	31 (33.7)	3 (6.3)

¹Choledochal cysts, transient cholestasis of unknown origin, and neonatal intrahepatic cholestasis caused by citrin deficiency were included as disease controls.

^aP < 0.05 vs healthy controls. Anti-MPO and/or anti-PR3: Any positivity of specific ANCAs; ANCAs: Any positivity of ANCAs by IIF or specific ANCA by ELISA. ANCA: Anti-neutrophil cytoplasmic antibody; BA: Biliary atresia; ELISA: Enzyme linked immunosorbent assay; IIF: Indirect immunofluorescence.

Table 5 Association of autoantibodies and prognosis of Kasai procedure with follow-up of 52 biliary atresia patients n (%)

Parameter	Anti-M2-3E		ANA		ANCA
	Positive, n = 17	Negative, n = 35	Positive, n = 8	Negative, n = 44	Positive, n = 29	Negative, n = 23
TB, μmol/L	105.5 ± 80.3	109.4 ± 106.8	41.6 ± 49.5	127.9 ± 135.0	133.0 ± 120.7	85.3 ± 66.2
ALT, U/L	184.1 ± 119.8	171.7 ± 121.4	127.9 ± 135.0	185.8 ± 115.7	171.46 ± 95.5	179.6 ± 140.3
Occurrence of cholangitis	9 (52.9)	20 (57.1)	5 (62.5)	24 (54.5)	24 (82.8)a	5 (21.7)a

Data are described as mean ± SD.

^aP < 0.05, r = 0.61; ANCA showed a positive correlation to the occurrence of postoperative cholangitis. TB is showed as a parameter of persistence of jaundice (TB > 34 μmol/L); ALT is showed as a parameter of acute liver injury (ALT > 35 U/L). ANCAs: Any positivity of ANCAs or specific ANCAs. ALT: Alanine aminotransferase; ANA: Anti-nuclear antibody; ANCA: Anti-neutrophil cytoplasmic antibody; BA: Biliary atresia; TB: Total bilirubin.