HLA variants related to primary sclerosing cholangitis influence rejection after liver transplantation

Table 1 Distribution and frequency of the underlying liver disease of 143 liver transplant recipients

Indication for transplantation	n (%)
Autoimmune etiology	65 (45.5)
Primary sclerosing cholangitis	44 (30.8)
Primary biliary cirrhosis	14 (9.8)
Autoimmune hepatitis	7 (4.9)
Viral hepatitis	20 (13.9)
Hepatitis B	5 (3.5)
Hepatitis C	15 (10.5)
Other indications	65 (43.9)
Hepatocellular carcinoma	13 (8.8)
Secondary liver tumours	7 (4.7)
Acute hepatic failure	8 (5.4)
Alcoholic cirrhosis	17 (12.2)
Cholangiocarcinoma	10 (6.8)
Budd-Chiari	2 (1.4)
Cryptogenic cirrhosis	9 (6.1)
Metabolic liver disease	1 (0.5)
Polycystic liver disease	2 (1.4)

Table 2 Frequencies of human leukocyte antigen-B alleles in the acute cellular rejection group (n = 40) and the non-acute cellular rejection group (n =102) in the total population (missing n = 1)

HLA-B allele	Acute rejection		No acute rejection		OR	95%CI	Uncorrected
	n (alleles)	(%)	n (alleles)	(%)			P value
*05	0	(0)	8	(4)	0.1	0.0-1.1	0.070
*07	10	(13)	23	(11)	1.2	0.6-2.5	0.690
*08	27	(34)	34	(17)	2.5	1.4-4.6	0.002
*12	12	(14)	27	(13)	1.2	0.6-2.3	0.700
*13	1	(1)	5	(2)	1.1	0.3-4.6	1.000
*14	2	(2)	4	(2)	1.1	0.3-4.6	1.000
*15	6	(7)	28	(14)	0.5	0.2-1.2	0.150
*16	1	(1)	4	(2)	0.7	0.1-3.4	0.510
*17	1	(1)	6	(3)	0.6	0.1-2.8	0.410
*18	1	(1)	6	(3)	0.4	0.1-2.0	0.240
*21	0	(0)	2	(1)	0.5	0.1-4.8	0.360
*22	3	(3)	2	(1)	3.6	0.8-16.4	0.120
*27	4	(5)	10	(5)	1.1	0.4-3.2	1.000
*35	4	(6)	19	(9)	0.7	0.3-1.8	0.230
*37	4	(5)	4	(2)	2.6	0.7-9.1	0.180
*40	4	(5)	19	(9)	0.6	0.2-1.5	0.220
*41	1	(1)	2	(1)	1.5	0.3-9.1	0.870
*47	0	(0)	1	(0.5)	0.8	0.1-9.2	0.530

OR and corresponding 95%CI were calculated using Woolf’s formula with Haldane’s correction; uncorrected P value: calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen; AR: Acute cellular rejection.

Table 3 Frequencies of human leukocyte antigen-C alleles in the acute cellular rejection group (n = 40) and the non-acute cellular rejection group (n = 102) in the total population (missing n = 1)

HLA-C allele	Acute rejection		No acute rejection		OR	95%CI	Uncorrected
	n (alleles)	(%)	n (alleles)	(%)			P value
*01	2	(2.5)	8	(3.9)	0.6	0.3-2.9	0.43
*02	2	(2.5)	11	(5.4)	0.5	0.1-1.8	0.24
*03	10	(12.5)	43	(21.1)	0.7	0.3-1.1	0.09
*04	8	(10.0)	24	(11.8)	1.1	0.4-2.2	0.89
*05	8	(10.0)	18	(8.8)	1.0	0.5-2.3	0.76
*06	6	(7.5)	16	(7.8)	1.2	0.5-2.8	0.82
*07	41	(51.2)	62	(30.4)	2.4	1.4-4.0	0.001
*08	2	(2.5)	4	(2.0)	1.1	0.3-4.7	0.54
*12	0	(0)	8	(4.2)	0.2	0.02-1.1	0.07
*14	0	(0)	1	(0.5)	0.9	0.08-9.4	0.72
*15	0	(0)	4	(2.0)	0.3	0.03-2.5	0.26
*16	0	(0)	3	(1.5)	0.4	0.04-3.2	0.31
*17	1	(1.2)	4	(2.0)	0.8	0.2-4.3	0.57

OR and corresponding 95%CI were calculated using Woolf’s formula with Haldane’s correction; uncorrected P value: calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen.

Table 4 Frequencies of human leukocyte antigen-DRB1 alleles in the acute cellular rejection group (n = 40) and the non-AR group (n = 103) in the total population

HLA-DRB1	Acute rejection		No acute rejection		OR	95%CI	Uncorrected
allele	n (alleles)	(%)	n (alleles)	(%)			P value
*01	10	(12.5)	23	(11.2)	1.2	0.5-2.5	0.75
*02	15	(18.8)	31	(15.0)	1.3	0.7-2.6	0.44
*03	25	(31.3)	40	(19.4)	1.9	1.0-3.3	0.03
*04	4	(5.0)	46	(22.3)	0.2	0.1-0.5	0.001
*07	7	(8.8)	16	(7.8)	1.2	0.5-2.8	0.78
*08	2	(2.5)	12	(5.8)	0.5	0.1-1.8	0.20
*09	0	(0)	1	(0.5)	0.9	0.1-9.5	0.72
*10	1	(1.3)	4	(1.9)	0.8	0.2-4.4	0.57
*11	1	(1.3)	6	(2.9)	0.6	0.1-2.9	0.37
*12	1	(1.3)	3	(1.5)	1.1	0.2-6.1	0.69
*13	13	(16.3)	21	(10.2)	1.7	0.8-3.6	0.16
*14	1	(1.3)	3	(1.5)	1.1	0.2-6.1	0.69

OR and corresponding 95%CI were calculated using Woolf’s formula with Haldane’s correction; uncorrected P value is calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen.

Table 5 Comparison of the frequencies of human leukocyte antigen-B alleles in the acute cellular rejection group (n = 23) and the non-acute cellular rejection group (n = 76) in the non-primary sclerosing cholangitis population

HLA-B	Acute rejection		Non acute rejection		OR	95%CI	Uncorrected
allele	n (alleles)	(%)	n (alleles)	(%)			P value
*05	0	(0)	4	(2.6)	0.3	0.1-2.9	0.34
*07	6	(13.0)	15	(9.9)	1.4	0.5-3.1	0.54
*08	12	(26.0)	20	(13.2)	2.1	0.8-4.6	0.04
*12	9	(19.6)	20	(13.2)	1.6	0.6-3.4	0.36
*13	1	(2.2)	4	(2.6)	1.0	0.2-5.4	0.67
*14	2	(4.3)	4	(2.6)	1.8	0.5-5.81	0.43
*15	5	(10.9)	21	(13.8)	0.8	0.3-2.1	0.51
*16	1	(2.2)	3	(2.6)	1.0	0.2-5.34	1.00
*17	0	(0)	4	(2.6)	0.3	0.1-2.9	0.63
*18	1	(2.2)	6	(3.9)	0.6	0.1-2.0	1.00
*21	0	(0)	2	(1.3)	0.7	0.1-2.5	1.00
*22	0	(0)	2	(1.3)	0.7	0.11-2.5	0.71
*27	2	(4.3)	7	(4.6)	1.0	0.3-4.0	1.00
*35	4	(8.7)	15	(9.9)	0.8	0.3-2.5	1.00
*37	2	(4.3)	3	(1.9)	2.4	0.4-10.3	0.34
*40	2	(4.3)	17	(11.2)	0.4	0.1-1.4	0.25
*41	1	(2.2)	2	(1.3)	1.9	0.3-11.3	0.56
*47	0	(0)	1	(0.7)	1.0	0.1-11.4	1.00

OR and corresponding 95%CIs were calculated using Woolf’s formula with Haldane’s correction. Uncorrected P value is calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen.

Table 6 Comparison of the frequencies of human leukocyte antigen-C alleles in the acute cellular rejection group (n = 23) and the non-acute cellular rejection group (n = 76) in the non-primary sclerosing cholangitis population

HLA-C	Acute rejection		No acute rejection		OR	95%CI	Uncorrected
allele	n (alleles)	(%)	n (alleles)	(%)			P value
*01	1	(2.2)	5	(3.3)	0.9	0.16-4.27	1.00
*02	1	(2.2)	7	(4.6)	0.6	0.11-2.53	0.68
*03	4	(8.7)	34	(22.4)	0.4	0.13-0.93	0.06
*04	7	(15.2)	21	(13.8)	1.2	0.45-2.61	0.81
*05	6	(13.0)	12	(7.9)	1.8	0.55-3.74	0.38
*06	3	(6.5)	12	(7.9)	0.9	0.27-2.74	1.00
*07	23	(50.0)	40	(26.3)	2.8	1.42-5.42	0.003
*08	2	(4.3)	4	(2.6)	1.9	0.34-5.89	0.63
*12	0	(0)	6	(3.9)	0.2	0.02-1.62	0.34
*15	0	(0)	4	(2.6)	0.4	0.04-2.94	0.58
*16	0	(0)	3	(2.0)	0.5	0.05-3.97	0.32
*17	1	(2.2)	4	(2.6)	1.1	0.19-5.46	1.00

OR and corresponding 95%CI were calculated using Woolf’s formula with Haldane’s correction. Uncorrected P value calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen.

Table 7 Comparison of the frequencies of human leukocyte antigen-DRB1 alleles in the acute cellular rejection group (n = 23) and the non-acute cellular rejection group (n = 76) in the non- primary sclerosing cholangitis population

HLA-DRB1	Acute rejection		No acute rejection		OR	95%CI	Uncorrected
allele	n (alleles)	(%)	n (alleles)	(%)			P value
*01	7	(15.2)	18	(11.8)	1.4	0.6-3.4	0.55
*02	8	(17.4)	23	(15.1)	1.2	0.5-2.8	0.46
*03	13	(28.2)	24	(15.8)	2.5	1.9-5.4	0.05
*04	3	(6.5)	35	(23.0)	0.3	0.1-0.9	0.02
*07	6	(13.0)	12	(7.9)	1.8	0.7-4.8	0.29
*08	2	(4.3)	10	(6.6)	0.7	0.2-2.7	0.73
*10	0	(0)	3	(2.0)	0.5	0.1-4.2	0.59
*11	1	(2.2)	6	(3.9)	0.7	0.1-3.7	1.00
*12	1	(2.2)	3	(2.0)	1.4	0.2-7.9	0.93
*13	7	(15.2)	15	(9.9)	1.7	0.7-4.2	0.31
*14	0	(0)	3	(2.0)	0.5	0.1-4.2	1.00

OR and corresponding 95%CIs were calculated using Woolf’s formula with Haldane’s correction. Uncorrected P value is calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen.

Table 8 Data on human leukocyte antigen mismatches on the risk of acute cellular rejection after liver transplantation n (%)

Locus	MM (n)	Non-AR group (n = 102)	AR group (n = 41)	P value
HLA-A	1-2	86 (84.3)	38 (92.7)	0.18
	0	16 (15.7)	3 (7.3)
HLA-B	1-2	97 (95.1)	41 (100)	0.15
	0	5 (4.9)	0 (0)
HLA-DR	1-2	94 (92.2)	40 (97.6)	0.21
	0	8 (7.8)	1 (2.4)
A, B, DR	0-4	62 (60.8)	19 (47.3)	0.12
	5-6	40 (39.2)	22 (53.7)

Uncorrected P values were calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen; MM: Mismatches; AR: Acute cellular rejection.

Table 9 Data on human leukocyte antigen mismatches on the risk of acute cellular rejection after liver transplantation, according to primary sclerosing cholangitis or non-primary sclerosing cholangitis n (%)

Locus	MM	Non-AR group	AR group	P value
	(n)	(PSC, n = 26)	(PSC) n = 17)
		(Non-PSC, n = 76)	(Non-PSC) n = 24)
HLA-A
(PSC)	1-2	20 (57.1)	15 (42.9)	0.35
	0	6 (75.0)	2 (25.0)
(non-PSC)	1-2	65 (73.9)	23 (26.1)	0.18
	0	11 (92.7)	1 (7.3)
HLA-B
(PSC)	1-2	25 (59.5)	17 (40.5)	0.41
	0	1 (100.0)	0 (0.0)
(non-PSC)	1-2	73 (75.3)	24 (24.7)	0.32
	0	3 (100.0)	0 (0.0)
HLA-DR
(PSC)	1-2	26 (61.9)	16 (38.1)	0.44
	0	0 (0.0)	1 (100.0)
(non-PSC)	1-2	67 (74.4)	23 (25.6)	0.27
	0	9 (90.0)	1 (10.0)
A, B, DR
(PSC)	5-6	12 (54.5)	10 (45.5)	0.42
	0-4	14 (66.7)	7 (33.3)
(non-PSC)	5-6	29 (67.4)	14 (32.6)	0.08
	0-4	47 (82.4)	10 (17.6)

Uncorrected P values were calculated by the χ² test or the Fisher’s exact test where appropriate. HLA: Human leukocyte antigen; PSC: Primary sclerosing cholangitis; MM: mismatches; AR: acute cellular rejection.

Table 10 Data on the relationship between killer immunoglobulin-like receptor gene phenotype (presence/absence of gene) in the recipient and the risk of acute cellular rejection after liver transplantation in the total patient population

Recipient KIR gene phenotype		Incidence of AR	P value
2DL1	Negative (n = 3)	0%	0.37
	Positive (n = 140)	29%
2DL2	Negative (n = 79)	28%	0.92
	Positive (n = 64)	29%
2DL3	Negative (n = 10)	20%	0.44
	Positive (n = 133)	28%
2DL4	Negative (n = 0)	-	-
	Positive (n = 143)	29%
2DL5	Negative (n = 79)	32%	0.27
	Positive (n = 64)	24%
2DP1	Negative (n = 3)	0%	0.54
	Positive (n = 140)	31%
2DS1	Negative (n = 91)	32%	0.24
	Positive (n = 52)	20%
2DS2	Negative (n = 76)	27%	0.85
	Positive (n = 67)	29%
2DS3	Negative (n = 108)	28%	0.79
	Positive (n = 35)	31%
2DS4DEL1	Negative (n = 23)	9%	0.03
	Positive (n = 120)	31%
2DS4WT2	Negative (n = 77)	27%	0.84
	Positive (n = 66)	29%
2DS4TOTA3	Negative (n = 3)	0%	0.67
	Positive (n = 140)	29%
2DS5	Negative (n = 104)	32%	0.47
	Positive (n = 39)	24%
3DL1E44	Negative (n = 3)	0%	0.61
	Positive (n = 140)	29%
3DL1E95	Negative (n = 3)	0%	0.62
	Positive (n = 140)	28%
3DL2	Negative (n = 0)	-	-
	Positive (n = 143)	29%
3DL3	Negative (n = 0)	-	-
	Positive (n = 143)	29%
3DP1	Negative (n = 0)	-	-
	Positive (n = 143)	29%
3DS1	Negative (n = 91)	31%	0.39
	Positive (n = 52)	25%

¹2DS4DEL refers to the 22-bp deletion variant of 2DS4;

²2DS4WT refers to the full-length form of the gene;

³2DS4TOT refers to the total number of 2DS4, i.e., 2DS4DEL and 2DS4WT combined; ^4,53DL1E4 and 3DL1E9 refers to exon 4 and 9 of the 3DL1 gene respectively. The various KIR genes genotyped are listed in the leftmost column. Genes with S in the name (e.g., KIR2DS4) encode activating KIRs, genes with L in the name (e.g., KIR3DL1) encode inhibiting KIRs and genes with a P in the name (e.g., KIR3DP1) are pseudogenes. Uncorrected P value is calculated by the χ² test or the Fisher’s exact test where appropriate. KIR: Killer immunoglobulin-like receptor; AR: Acute cellular rejection.

Table 11 Data on the relationship between killer immunoglobulin-like receptor gene phenotype in the recipient and the risk of acute cellular rejection after liver transplantation in patients with primary sclerosing cholangitis compared with patients without primary sclerosing cholangitis

Recipient KIR gene phenotype	Incidence of AR	P value
2DL1
(PSC)		-
Negative (n = 0)	-
Positive (n = 4)	40%
(other)		1.00
Negative (n = 3)	0%
Positive (n = 96)	25%
2DL2
(PSC)		0.37
Negative (n = 23)	33%
Positive (n =21)	48%
(other)		0.71
Negative (n = 54)	26%
Positive (n = 45)	22%
2DL3
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		1.00
Negative (n = 10)	20%
Positive (n = 89)	24%
2DL4
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		-
Negative (n = 0)	-
Positive (n = 99)	24%
2DL5
(PSC)		0.17
Negative (n = 28)	48%
Positive (n = 16)	25%
(other)		0.81
Negative (n = 51)	25%
Positive (n = 48)	22%
2DP1
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		1.00
Negative (n = 3)	0%
Positive (n = 96)	25%
2DS1
(PSC)		0.14
Negative (n = 13)	26%
Positive (n = 31)	20%
(other)		0.51
Negative (n = 60)	26%
Positive (n = 39)	20%
2DS2
(PSC)		0.37
Negative (n = 23)	33%
Positive (n = 21)	48%
(other)		0.48
Negative (n = 54)	26%
Positive (n = 45)	20%
2DS3
(PSC)		0.78
Negative (n = 34)	41%
Positive (n = 11)	36%
(other)		0.57
Negative (n = 76)	22%
Positive (n = 25)	28%
2DS4DEL1
(PSC)		0.11
Negative (n = 8)	38%
Positive (n = 36)	43%
(other)		0.10
Negative (n = 15)	7%
Positive (n = 85)	27%
2DS4WT2
(PSC)		0.81
Negative (n = 21)	38%
Positive (n = 23)	42%
(other)		0.71
Negative (n = 56)	22%
Positive (n = 43)	26%
2DS4TOTA3
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		1.00
Negative (n = 3)	0%
Positive (n = 96)	25%
2DS5
(PSC)		0.46
Negative (n = 34)	43%
Positive (n = 10)	30%
(other)		0.56
Negative (n = 69)	25%
Positive (n = 30)	20%
3DL1E44
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		-
Negative (n = 0)	-
Positive (n = 99)	25%
3DL1E95
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		1.00
Negative (n = 3)	0%
Positive (n = 96)	25%
3DL2
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		-
Negative (n = 0)	-
Positive (n = 98)	24%
3DL3
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		-
Negative (n = 0)	-
Positive (n = 99)	24%
3DP1
(PSC)		-
Negative (n = 0)	-
Positive (n = 44)	40%
(other)		-
Negative (n = 0)	-
Positive (n = 99)	24%
3DS1
(PSC)		0.42
Negative (n = 31)	44%
Positive (n =13)	31%
(other)		0.81
Negative (n = 61)	25%
Positive (n = 40)	23%

¹2DS4DEL refers to the 22-bp deletion variant of 2DS4;

²2DS4WT refers to the full-length form of the gene;

³2DS4TOT refers to the total number of 2DS4, e.g., 2DS4DEL and 2DS4WT combined; ^4,53DL1E4 and 3DL1E9 refers to exon 4 and 9 of the 3DL1 gene respectively. The various KIR genes genotyped are listed in the leftmost column. Genes with S in the name (e.g., KIR2DS4) encode activating KIRs, genes with L in the name (e.g., KIR3DL1) encode inhibiting KIRs and genes with a P in the name (e.g., KIR3DP1) are pseudogenes. Uncorrected P value was calculated by the χ² test or the Fisher’s exact test where appropriate. KIR: Killer immunoglobulin-like receptor; AR: Acute cellular rejection; PSC: Primary sclerosing cholangitis.

Table 12 Killer immunoglobuline-like receptor gene copy number of the recipient and acute cellular rejection after liver transplantation

KIR gene	AR	Non-AR	P value
	mean copy number (95% CI)	mean copy number (95%CI)
KIR2DL1	1.69 (1.54-1.84)	1.64 (1.53-1.76)	0.64
KIR2DL2	0.52 (0.34-0.71)	0.53 (0.40-0.65)	0.97
KIR 2DL3	1.50 (1.30-1.70)	1.45 (1.33-1.58)	0.69
KIR 2DL4	2.10 (2.00-2.19)	1.98 (1.91-2.05)	0.06
KIR 2DL5	0.50 (0.27-0.73)	0.63 (0.49-0.78)	0.32
KIR 2DP1	1.71 (1.57-1.86)	1.70 (1.59-1.81)	0.89
KIR 2DS1	0.26 (0.12-0.40)	0.43 (0.33-0.54)	0.08
KIR 2DS2	0.62 (0.30-0.43)	0.54 (0.41-0.66)	0.56
KIR 2DS3	0.29 (0.13-0.44)	0.32 (0.20-0.40)	0.77
KIR 2DS4DEL1	1.17 (1.01-1.32)	1.08 (0.94-1.21)	0.45
KIR 2DS4WT2	0.55 (0.36-0.73)	0.48 (0.37-0.59)	0.53
KIR 2DS4TOT3	1.74 (1.60-1.88)	1.56 (1.45-1.67)	0.06
KIR 2DS5	0.21 (0.08-0.35)	0.32 (0.22-0.42)	0.25
KIR 3DL1E44	1.74 (1.60-1.88)	1.57 (1.46-1.67)	0.08
KIR 3DL1E95	1.74 (1.60-1.88)	1.56 (1.46-1.67)	0.06
KIR 3DL2	-	-	-
KIR 3DL3	-	-	-
KIR 3DP1	-	-	-
KIR 3DS1	0.33 (0.30-0.51)	0.40 (0.30-0.51)	0.47

The various KIR genes genotyped are listed in the leftmost column. Genes with S in the name (e.g., KIR2DS4) encode activating KIRs, genes with L in the name (e.g., KIR3DL1) encode inhibiting KIRs and genes with a P in its name (e.g., KIR3DP1) are pseudogenes. Statistical comparison was performed using the independent sample t test.

¹2DS4DEL refers to the 22-bp deletion variant of 2DS4;

²2DS4WT refers to the full-length form of the gene;

³2DS4TOT refers to the total number of 2DS4, i.e., 2DS4DEL and 2DS4WT combined; ^4,53DL1E4 and 3DL1E9 refers to exon 4 and 9 of the 3DL1 gene respectively. KIR: Killer immunoglobulin-like receptor; AR: Acute cellular rejection.

Table 13 Data showing scarce, but similar numbers of natural killer cells and natural killer T cells in biopsies with equal grade of AR in patients with various primary liver diseases

MNC cell subset1	PSC (n = 10)	PBC (n = 10)	Non-imm (n = 10)	P value
CD56 (NK/NKT)	1.6 ± 0.3	1.8 ± 0.4	1.8 ± 0.3	0.9
CD57 (NK/NKT)	5.0 ± 0.9	4.0 ± 0.4	4.6 ± 0.6	0.1
CD3-CD56+ (NK)	0.2 ± 0.1	0.2 ± 0.1	0.3 ± 0.1	0.7
CD3-CD57+ (NK)	0	0	0	-

¹Mean absolute number of NK/NKT cells/mm². Statistics were performed comparing mean value between groups using one-way ANOVA. MNC: Mononuclear cells; PSC: Primary sclerosing cholangitis; PBC: Primary biliary cirrhosis; Non-imm: Non-immunological disease; NKT: Natural killer T cell; NK: Natural killer.