Clinicopathological study of primary biliary cirrhosis with interface hepatitis compared to autoimmune hepatitis

Table 1 Main clinicopathological features of primary biliary cirrhosis with interface hepatitis and autoimmune hepatitis

	PBC1 (n = 41)	AIH (n = 43)
Gender (male:female)	5:36	4:39
Age (yr, mean ± SD)	59.0 ± 13.3	61.7 ± 10.8
Laboratory data
Ratio of AMA(+) or M2(+) and titer of AMA	75% and 141.2 ± 211.4	7% and 0 ± 0
Ratio of ANA(+) and titer2	72% and 724.3 ± 1262.4	91% and 411.1 ± 550.8
AST (IU/L)	51.6 ± 29.7	219.5 ± 180.5
ALT (IU/L)	54.0 ± 40.5	261.8 ± 253.4
ALP (IU/L)	615.5 ± 572.8	449.9 ± 251.8
IgG (mg/dL)	1832.9 ± 607.2	2246.2 ± 673.0
IgM (mg/dL)	300.9 ± 220.6	193.7 ± 120.0
IgA (mg/dL)	307.9 ± 122.4	387.2 ± 183.1

Laboratory data is shown as the mean ± SD; anti-mitochondrial antibodies (AMA)(+) ≥ 40; M2(+) ≥ 5; antinuclear antibodies (ANA)(+) ≥ 40.

¹With interface hepatitis;

²The average titer of ANA was higher in primary biliary cirrhosis (PBC) than in autoimmune hepatitis (AIH) because two PBC cases had a titer of 5120, which influenced the average. AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; ALP: Alkaline phosphatase.

Table 2 Primary antibodies used in this study

Primary antibody	Source	Clone	Animal	Type	Pretreatment	Dilution
CD3	Nichirei	PS1	Mouse	Mono	MW3	1:1
CD4	Nichirei	4B12	Mouse	Mono	MW3	1:1
CD8	Dako	C8/144B	Mouse	Mono	MW1	1:20
CD20	Dako	L26	Mouse	Mono	-	1:400
CD38	Epitomics	EPR4106	Rabbit	Mono	AC2	1:800
CD138	Dako	ML15	Mouse	Mono	MW3	1:100
IgG	Dako	A57H	Mouse	Mono	-	1:200
IgM	Nichirei	R1/69	Mouse	Mono	MW3	1:1
IgA	Novocastra	N1CLA	Mouse	Mono	PK	1:300

¹Citrate, pH = 6.0;

²Ethylenediaminetetraacetic acid (EDTA) buffer, pH8.0;

³ Tris EDTA buffer, pH = 9.0. MW: Microwave; AC: Autoclave; PK: Proteinase K, pH = 7.5 (Dako).

Table 3 Degree of mononuclear cell infiltration with respect to immunological phenotypes at the interface and in the hepatic lobules

	PBC1	AIH	P value
At the interface; scores of infiltrating mononuclear cells positive for
CD3	2.8 ± 1.2 (3)2	4.1 ± 1.1 (4)	0.0002
CD4	1.5 ± 1.0 (1)	2.0 ± 1.2 (2)	0.0387
CD8	1.5 ± 0.9 (1)	2.7 ± 1.7 (3)	0.0048
CD20	0.7 ± 0.6 (1)	1.0 ± 1.0 (1)	0.0046
CD38	2.7 ± 1.5 (3)	3.7 ± 1.3 (4)	0.0046
IgG	1.3 ± 1.0 (1)	1.8 ± 1.3 (2)	0.0301
IgM	1.0 ± 0.8 (1)	0.6 ± 0.8 (0.5)	0.0249
IgA	0.5 ± 0.6 (0)	0.6 ± 0.7 (1)	0.4486
In the hepatic lobules; scores of infiltrating mononuclear cells positive for
CD3	2.2 ± 0.9 (2)	3.3 ± 1.3 (3)	0.0016
CD4	0.8 ± 0.9 (0.5)	1.8 ± 1.1 (2)	0.0002
CD8	0.8 ± 0.8 (1)	2.5 ± 1.5 (2)	< 0.0001
CD20	0.3 ± 0.6 (0)	0.7 ± 0.8 (0.5)	0.0147
CD38	1.1 ± 1.0 (1)	2.0 ± 1.2 (2)	0.0025
IgG	0.6 ± 0.8 (0)	1.4 ± 1.3 (1)	0.0007
IgM	0.4 ± 0.5 (0)	0.6 ± 0.7 (0.5)	0.166
IgA	0.3 ± 0.7 (0)	0.6 ± 0.7 (0.5)	0.0239

¹With interface hepatitis;

²The scores were graded from 0-6 for each case, and are shown as the mean ± SD and (median). PBC: Primary biliary cirrhosis; AIH: Autoimmune hepatitis.

Table 4 Clinicopathological features of primary biliary cirrhosis based on the predominant immunoglobulin class of the plasma cell

No. of cases(female:male)	Group A28 (25:3)	Group B12 (10:2)	P value
Age (yr)	56.1 ± 14.5 (56)	66.0 ± 7.2 (66)	0.0479
Laboratory data
AST (IU/L)	50.5 ± 31.2 (43.5)	55.0 ± 28.3 (42.5)	0.5839
ALT (IU/L)	54.2 ± 43.6 (45.5)	55.0 ± 35.4 (46.5)	0.7238
ALP (IU/L)	681.7 ± 655.0 (446)	389.1 ± 136.3 (358.5)	0.4166
IgG (mg/dL)	1737.4 ± 510.4 (1600)	2081.3 ± 784.4 (2062.5)	0.2658
IgM (mg/dL)	340.78 ± 219.0 (280.5)	197.3 ± 198.7 (133.5)	0.0077
IgA (mg/dL)	309.2 ± 129.8 (281)	304.6 ± 107.0 (268.5)	0.8184
AMA, M2 (positive ratio)	80.10%	55.60%	0.1916
ANA (positive ratio)	65.40%	80.00%	0.6880
Histological findings (scores)
Interface hepatitis1	2.4 ± 0.6 (2)	2.8 ± 0.6 (3)	0.0688
Lobular hepatitis2	2.3 ± 0.6 (2)	2.3 ± 0.6 (2)	0.8577
Infiltration of mononuclear cells positive for (scores)
CD3 at the interface3	2.7 ± 1.2 (2.5)	2.8 ± 1.3 (3)	0.7039
CD38 at the interface3	2.4 ± 1.5 (2)	3.3 ± 1.3 (4)	0.0205
CD3 in the hepatic lobules3	2.1 ± 0.8 (2)	2.4 ± 1.2 (2)	0.5341
CD38 in the hepatic lobules3	0.9 ± 0.9 (1)	1.6 ± 1.1 (2)	0.0497

¹The score was graded from 2 to 4 for each case;

²the score was graded from 0 to 4 for each case;

³ The score was graded from 0 to 6 for each case. Group A, IgM-predominant cases and IgM/IgG-equal cases; group B, IgG-predominant cases. Age, laboratory data expect autoantibodies and scores are shown as the mean ± SD and (median); anti-mitochondrial antibodies (AMA)(+) ≥ 40; M2(+) ≥ 5; antinuclear antibodies (ANA)(+) ≥ 40. AST: Aspartate aminotransferase; ALT: Alanine aminotransferase; ALP: Alkaline phosphatase.

Table 5 Correlation between the elevations in aspartate aminotransferase and the scores of hepatitis and mononuclear cell infiltration of primary biliary cirrhosis and autoimmune hepatitis

Scores of inflammatory cells at the interface vs elevation of aspartate aminotransferase	Primary biliary cirrhosis1		Autoimmune hepatitis
	r	P value	r	P value
Scores of interface hepatitis	0.3600	0.0286	0.1427	0.3796
Scores of lobular hepatitis	0.4056	0.0128	0.4444	0.0041
Scores of CD3+ cells at the interface	0.4172	0.0114	0.1540	0.4724
Scores of CD3+ in the lobules	0.3487	0.0371	0.4306	0.0357
Scores of CD4+ at the interface	0.4410	0.0071	-0.1122	0.6017
Scores of CD4+ in the lobules	0.4478	0.0062	0.3033	0.1496
Scores of CD8+ at the interface	0.3466	0.0384	0.3051	0.1471
Scores of CD8+ in the lobules	0.1802	0.2929	0.7074	0.0001
Scores of CD38+ at the interface	0.6010	0.0001	0.0969	0.6601
Scores of CD38+ in the lobules	0.3821	0.0215	0.1541	0.4827
Scores of IgG+ at the interface	0.3909	0.0184	0.1381	0.5200
Scores of IgM+ at the interface	0.5436	0.0006	-0.1372	0.5225

¹With interface hepatitis.

Table 6 Comparison of the degree of hepatitis and mononuclear cell infiltration in autoantibody positive and negative groups in primary biliary cirrhosis and autoimmune hepatitis

PBC	AMA(+) or M2(+)	AMA(-) and M2(-)	P value
Comparison between AMA , M2 positive and negative group in PBC
Scores of interface hepatitis1	2.41 ± 0.64 (2)	2.33 ± 0.50 (2)	0.9120
Scores of CD3+ cells at interface2	2.62 ± 1.13 (2)	2.44 ± 1.24 (3)	0.7552
Scores of CD38+ cells at interface2	2.42 ± 1.39 (2.5)	2.33 ± 1.22 (2)	0.9843
Scores of IgG+ cells at interface2	1.12 ± 0.59 (1)	1.22 ± 1.64 (1)	0.4977
Scores of IgM+ cells at interface2	1.00 ± 0.63 (1)	0.89 ± 0.93 (1)	0.4196
Comparison between ANA positive and negative group in PBC
Scores of interface hepatitis1	2.42 ± 0.65 (2)	2.30 ± 0.48 (2)	0.6241
Scores of CD3+ cells at interface2	2.47 ± 1.16 (2)	2.80 ± 1.23 (2)	0.4548
Scores of CD38+ cells at interface2	2.43 ± 1.16 (2)	2.50 ± 1.18 (3)	0.8113
Scores of IgG+ cells at interface2	1.09 ± 1.08 (1)	1.30 ± 0.67 (1)	0.4193
Scores of IgM+ cells at interface2	0.83 ± 0.65 (1)	1.30 ± 0.82 (1)	0.0742
Comparison between ANA positive and negative group in AIH
Scores of interface hepatitis1	2.70 ± 0.64 (3)	2.33 ± 0.58 (2)	0.3543
Scores of CD3+ cells at interface2	4.21 ± 1.13 (4)	3.33 ± 1.53 (3)	0.2965
Scores of CD38+ cells at interface2	3.89 ± 1.18 (4)	2.33 ± 1.15 (3)	0.0692
Scores of IgG+ cells at interface2	1.95 ± 1.39 (2)	1.00 ± 1.00 (1)	0.2552
Scores of IgM+ cells at interface2	0.63 ± 0.76 (1)	0.00 ± 0.00 (0)	0.1141

¹The scores were graded from 2 to 4 for each case, and are shown as the mean ± SD and (median);

²The scores were graded from 0 to 6 for each case, and are shown as the mean ± SD and (median). Anti-mitochondrial antibodies (AMA)(+) ≥ 40; M2(+) ≥ 5; antinuclear antibodies (ANA)(+) ≥ 40. PBC: Primary biliary cirrhosis; AIH: Autoimmune hepatitis.