Autoantibodies in Chinese patients with chronic hepatitis B: Prevalence and clinical associations

Table 1 Demographic and clinical features, biochemical parameters and response to antivirus treatment in chronic hepatitis B

	All patients (n = 325)
Male/female	201/124
Age (yr)	43.44 ± 16.19
Associated immune disease1n (%)	8 (2.4)
Cirrhosis	206 (63.4)
Hepatoma	38 (11.7)
Duration of HBsAg pos. n (%)	140 cases
1-5 yr	35 (25.0)
6-10 yr	25 (17.9)
11-15 yr	34 (24.3)
16-20 yr	28 (20.0)
> 20 yr	18 (12.9)
HBeAg2	147/266 (43.3)
AST (U/L)	145.10 ± 96.79
ALT (U/L)	126.51 ± 100.15
ALP (U/L)	121.08 ± 74.54
GGT (U/L)	73.43 ± 62.52
IgA (g/L)	3.24 ± 2.67
IgG (g/L)	14.73 ± 5.58
IgM (g/L)	1.37 ± 1.32
γ-globulin (g/L)	20.1 ± 7.38
Liver biopsy	31 cases
Staging and grading3n (%)
G1S1	5 (16.1)
G1S2	3 (9.7)
G1S3	1 (3.2)
G2S1	3 (9.7)
G2S2	6 (19.4)
G2S3	2 (6.5)
G2S4	2 (6.5)
G3S2	3 (9.7)
G3S3	4 (12.9)
G4S4	2 (6.5)
Anti-viral treatment response evaluation4n (%)	72 cases
Virological non-responders	6 (8.3)
Virological responders	66 (91.7)

¹Hashimoto’s thyroiditis and type 1 diabetes mellitus: 1 case each; hypothyroidism: 2 cases; probable autoimmune hepatitis: 4 cases;

²Hepatitis B e-antigen statuses were identified in 266 cases;

³The Scheuer system was used to score necroinflammatory activity and fibrosis/cirrhosis;

⁴The virological response was defined as hepatitis B virus DNA concentration of less than 2000 IU/mL at 12 mo after nucleoside/nucleotide therapy. AST: Aspartate aminotransferase; ALP: Alkaline phosphatase; GGT: γ-glutamyl transpeptidase; ALT: Alanine aminotransferase.

Table 2 Prevalence of the autoantibodies in chronic hepatitis B compared with chronic hepatitis C, autoimmune hepatitis, primary biliary cirrhosis, and healthy controls n (%)

	CHB (n = 325)		CHC (n = 71)		AIH (n = 11)		PBC (n = 71)		HC (n = 60)
	Pos.	P value	Pos.	P value	Pos.	P value	Pos.	P value	Pos.
Any autoantibody	189 (58.2)	0.211	47 (66.2)	0.004	11 (100.0)	< 0.001	71 (100.0)	< 0.001	4 (6.7)
ANA (IFA)	76 (23.4)	0.353	13 (18.3)	0.044	6 (54.5)	< 0.001	47 (66.2)	< 0.001	2 (3.3)
SMA (IFA)	10 (3.1)	1.000	2 (2.8)	1.000	0 (0.0)	0.220	0 (0.0)	0.373	0 (0.0)
AMA-M2 (LIA)	22 (6.8)	0.322	2 (2.8)	1.000	1 (9.1)	< 0.001	60 (84.5)	0.033	0 (0.0)
Anti-BPO (LIA)	23 (7.1)	0.124	1 (1.4)	1.000	1 (9.1)	< 0.001	60 (84.5)	0.034	0 (0.0)
Anti-Sp100 (LIA)	8 (2.5)	0.360	0 (0.0)	1.000	0 (0.0)	< 0.001	11 (15.5)	0.616	0 (0.0)
Anti-PML (LIA)	36 (11.1)	0.003	0 (0.0)	0.615	0 (0.0)	< 0.001	30 (42.3)	0.003	0 (0.0)
Anti-gp210 (LIA)	41 (12.6)	< 0.001	0 (0.0)	0.372	0 (0.0)	< 0.001	35 (49.3)	0.001	0 (0.0)
Anti-LKM-1 (LIA)	2 (0.6)	1.000	1 (1.4)	0.190	1 (9.1)	1.000	0 (0.0)	1.000	0 (0.0)
Anti-LC-1 (LIA)	10 (3.1)	0.707	1 (1.4)	0.810	1 (9.1)	0.901	3 (4.2)	0.373	0 (0.0)
Anti-SLA/LP (LIA)	7 (2.2)	0.360	0 (0.0)	1.000	0 (0.0)	0.555	3 (4.2)	0.602	0 (0.0)
Anti-Ro52 (LIA)	91 (28.0)	0.015	10 (14.1)	< 0.001	10 (90.9)	< 0.001	36 (50.7)	< 0.001	3 (5.0)

P < 0.05, CHB vs CHC, CHB vs AIH, CHB vs PBC, and CHB vs HC, respectively. CHB: Chronic hepatitis B; CHC: Chronic hepatitis C; AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis; HC: Health controls; ANA: Anti-nuclear antibodies; AMA: Anti-mitochondrial antibodies; SMA: Smooth-muscle antibodies; anti-PML: Anti-promyelocytic leukemia protein; anti-LKM-1: Anti-liver kidney microsome type 1; anti-LC-1: Anti-liver cytosolic antigen type 1; anti-SLA/LP: Anti-soluble liver antigen/liver pancreas; IFA: Indirect immunofluorescence assay; LIA: Line immunoassay.

Table 3 Autoantibody titers in chronic hepatitis B compared with chronic hepatitis C, autoimmune hepatitis, primary biliary cirrhosis, and healthy controls

	CHB (n = 325)		CHC (n = 71)		AIH (n = 11)		PBC (n = 71)		HC (n = 60)
	Pos. cases	Mean rank1	Pos. cases	Mean rank	Pos. cases	Mean rank	Pos. cases	Mean rank	Pos. cases	Mean rank
ANA	76	66.79a	13	50	6	97.5	47	83.57	2	100.5
SMA	10	6.20	2	8	0	-	0	-	0	-
AMA-M2	22	23.61d	2	11	1	11.0	60	51.71	0	-
Anti-BPO	23	22.85d	1	10.5	1	10.5	60	51.81	0	-
Anti-Sp100	8	8.44	0	-	1	5.0	11	12.50	0	-
Anti-PML	36	29.54b	0	-	3	19.0	30	43.15	0	-
Anti-gp210	41	28.49d	0	-	2	26.75	35	53.13	0	-
Anti-LKM-1	2	1.50	1	4	1	4.0	0	-	0	-
Anti-LC-1	10	9.45	1	7	3	10.17	3	7.00	0	-
Anti-SLA/LP	7	6.00	0	-	3	8.17	3	8.17	0	-
Anti-Ro52	91	68.89b	10	50.9	4	94.25	36	91.89	4	30.5

¹Kruskal-Wallis H test;

^aP < 0.05, Mann-Whitney U test vs the autoantibody titers with PBC;

^bP < 0.01, Mann-Whitney U test vs the autoantibody titers with PBC;

^dP < 0.01 Mann-Whitney U test vs the autoantibody titers with PBC; “-”: Not indicated; P < 0.05 was considered significant. CHB: Chronic hepatitis B; CHC: Chronic hepatitis C; AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis; HC: Health controls; ANA: Anti-nuclear antibodies; AMA: Anti-mitochondrial antibodies; SMA: Smooth-muscle antibodies; anti-PML: Anti-promyelocytic leukemia protein; anti-LKM-1: Anti-liver kidney microsome type 1; anti-LC-1: Anti-liver cytosolic antigen type 1; anti-SLA/LP: Anti-soluble liver antigen/liver pancreas; IFA: Indirect immunofluorescence assay; LIA: Line immunoassay.

Table 4 Prevalence of the autoantibodies in non-cirrhosis, compensated cirrhosis, and decompensated cirrhosis chronic hepatitis B groups n (%)

	Non-cirrhosis (n = 119)		Compensate cirrhosis (n = 85)		Decompensated cirrhosis (n = 121)
	Pos.	P value	Pos.	P value	Pos.	P value
ANA	39 (32.8)	0.006	16 (18.8)	0.027	21 (17.4)	0.787
SMA	1 (0.8)	0.393	3 (3.5)	0.131	6 (5.0)	0.882
AMA-M2	9 (7.6)	0.410	4 (4.7)	0.971	9 (7.4)	0.427
Anti-BPO	4 (3.4)	0.010	11 (12.9)	0.248	8 (6.6)	0.122
Anti-Sp100	2 (1.7)	1.000	2 (2.4)	0.694	4 (3.3)	1.000
Anti-PML	10 (8.4)	0.293	11 (12.9)	0.311	15 (12.4)	0.908
Anti-gp210	16 (13.4)	0.540	9 (10.6)	0.960	16 (13.2)	0.569
Anti-LKM-1	1 (0.8)	1.000	1 (1.2)	0.496	0 (0.0)	0.413
Anti-LC-1	3 (2.5)	1.000	3 (3.5)	1.000	4 (3.3)	1.000
Anti-SLA/LP	2 (1.7)	0.702	3 (3.5)	1.000	2 (1.7)	0.688
Anti-Ro52	33 (27.7)	0.629	21 (24.7)	0.628	37 (30.6)	0.356

P < 0.05, non-cirrhosis vs compensated cirrhosis, non-cirrhosis vs decompensated cirrhosis, and compensated vs decompensated cirrhosis, respectively. ANA: Anti-nuclear antibodies; AMA: Anti-mitochondrial antibodies; SMA: Smooth-muscle antibodies; anti-PML: Anti-promyelocytic leukemia protein; anti-LKM-1: Anti-liver kidney microsome type 1; anti-LC-1: Anti-liver cytosolic antigen type 1; anti-SLA/LP: Anti-soluble liver antigen/liver pancreas; IFA: Indirect immunofluorescence assay; LIA: Line immunoassay.

Table 5 Anti-nuclear antibodies fluorescence patterns and cirrhosis in chronic hepatitis B: Comparison of the non-cirrhosis, compensated cirrhosis and decompensated cirrhosis groups n (%)

	Non-cirrhosis1 (n = 117)		Compensated cirrhosis (n = 85)		Decompensated cirrhosis2 (n = 119)
	Pos.	P value	Pos.	P value	Pos.	P value
ANA-H	32 (27.4)	0.001	7 (8.2)	< 0.001	8 (6.7)	0.683
ANA-DM	1 (0.9)	0.403	3 (3.5)	0.223	5 (4.2)	1.000
ANA-other3	4 (3.4)	0.396	6 (7.1)	0.767	6 (5.0)	0.546

¹Two cases showed the combination of ANA-H/ANA-DM and ANA-DM/ANA-other fluorescence patterns, thus these two cases were excluded from this group;

²Two cases showed the combination of ANA-H/ANA-Others and ANA-DM/ANA-other fluorescence patterns, thus these two cases were excluded from this group;

³Fluorescence patterns such as centromeres, nuclear speckles, and nucleoli were included. P < 0.05, non-cirrhosis vs compensated cirrhosis, non-cirrhosis vs decompensated cirrhosis, and compensated vs decompensated cirrhosis, respectively, was considered significant. ANA: Anti-nuclear antibodies; ANA-H: ANA-homogeneous; ANA-DM: ANA-dots or membrane.

Table 6 Prevalence of the autoantibodies in the non-hepatocellular carcinoma and hepatocellular carcinoma chronic hepatitis B groups n (%)

	Non-HCC (n = 287)	HCC (n = 38)
	Pos.	Pos.	P value
ANA	68 (23.7)	8 (21.1)	0.718
SMA	10 (3.5)	0 (0.0)	0.613
AMA-M2	19 (6.6)	3 (7.9)	1.000
Anti-BPO	22 (7.7)	1 (2.6)	0.423
Anti-Sp100	8 (2.8)	0 (0.0)	0.603
Anti-PML	36 (12.5)	0 (0.0)	0.013
Anti-gp210	37 (12.9)	4 (10.5)	0.879
Anti-LKM-1	2 (0.7)	0 (0.0)	1.000
Anti-LC-1	10 (3.5)	0 (0.0)	0.613
Anti-SLA/LP	6 (2.1)	1 (2.6)	1.000
Anti-Ro52	82 (28.6)	9 (23.7)	0.528

ANA: Anti-nuclear antibodies; AMA: Anti-mitochondrial antibodies; SMA: Smooth-muscle antibodies; anti-PML: Anti-promyelocytic leukemia protein; anti-LKM-1: Anti-liver kidney microsome type 1; anti-LC-1: Anti-liver cytosolic antigen type 1; anti-SLA/LP: Anti-soluble liver antigen/liver pancreas.

Table 7 Dichotomization of autoantibodies into autoimmune hepatitis and primary biliary cirrhosis profiles by the specific autoantibody panels

	Autoantibody panel
AIH profile	ANA-H (IFA)
	SMA (IFA)
	Anti-LKM-1 (LIA)
	Anti-LC-1 (LIA)
	Anti-SLA/LP (LIA)
PBC profile	ANA-DM (IFA)
	AMA-M2 (LIA)
	Anti-BPO (LIA)
	Anti-Sp100 (LIA)
	Anti-PML (LIA)
	Anti-gp210 (LIA)

AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis; ANA: Anti-nuclear antibodies; SMA: Smooth-muscle antibodies; anti-PML: Anti-promyelocytic leukemia protein; anti-LKM-1: Anti-liver kidney microsome type 1; anti-LC-1: Anti-liver cytosolic antigen type 1; anti-SLA/LP: Anti-soluble liver antigen/liver pancreas. ANA-H: ANA-homogeneous; ANA-DM: ANA-dots or membrane; IFA: Indirect immunofluorescence assay; LIA: Line immunoassay.

Table 8 Autoimmune hepatitis and primary biliary cirrhosis profiles in chronic hepatitis B compared with chronic hepatitis C, autoimmune hepatitis, primary biliary cirrhosis, and healthy controls n (%)

	CHB (n = 325)		CHC (n = 71)		AIH (n = 11)		PBC (n = 71)		HC (n = 60)
	Pos.	P value	Pos.	P value	Pos.	P value	Pos.	P value	Pos.
AIH profile	43 (13.2)	0.655	8 (11.3)	0.083	4 (36.4)	0.655	8 (11.3)	0.003	0 (0)
PBC profile	71 (21.8)	< 0.001	2 (2.8)	0.954	3 (27.3)	< 0.001	63 (88.7)	< 0.001	0 (0)

P < 0.05, CHB vs CHC, CHB vs AIH, CHB vs PBC, and CHB vs HC, respectively. CHB: Chronic hepatitis B; CHC: Chronic hepatitis C; AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis; HC: Health controls.

Table 9 Autoimmune hepatitis and primary biliary cirrhosis profiles in the non-cirrhosis, compensated cirrhosis, and decompensated cirrhosis chronic hepatitis B groups n (%)

	Non-cirrhosis (n = 119)		Compensated cirrhosis (n = 85)		Decompensated cirrhosis (n = 121)
	Pos.	P1	Pos.	P2	Pos.	P3
AIH profile	22 (18.5)	0.039	7 (8.2)	0.133	14 (11.6)	0.436
PBC profile	17 (14.3)	0.038	22 (25.9)	0.019	32 (26.4)	0.928

P < 0.05, non-cirrhosis vs compensated cirrhosis, non-cirrhosis vs decompensated cirrhosis, and compensated vs decompensated cirrhosis, respectively. AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis.

Table 10 Autoimmune hepatitis and primary biliary cirrhosis profiles based on hepatitis B e-antigen status in chronic hepatitis B n (%)

	HBeAg pos.(n = 147)	HBeAg neg.(n = 119)	P value
AIH profile	23 (15.6)	7 (5.9)	0.012
PBC profile	24 (16.3)	29 (24.4)	0.102

P < 0.05, HBeAg positive group vs HBeAg negative group. AIH: Autoimmune hepatitis; PBC: Primary biliary cirrhosis; HBeAg: Hepatitis B e-antigen.