Supplementary Table 1. Number of CCL4L gene copies and risk of acute and chronic rejection in LT patient groups.

No acute rejection / Total acute rejection / p value / Odds Ratio / IC 95%
0-2 copies / 68 / 63 / 0.37 / 1.47 / (0.63-3.44)
≥ 3 copies / 11 / 15
No acute rejection / Acute rejection 1 episode / p value / Odds Ratio / IC 95%
0-2 copies / 68 / 49 / 0.60 / 1.32 / (0.46-3.81)
≥ 3 copies / 11 / 6
No acute rejection / Acute rejection > 1 episode / p value / Odds Ratio / IC 95%
0-2 copies / 68 / 14 / 0.011 / 3.97 / (1.39-11.38)
≥ 3 copies / 11 / 9
Acute rejection 1 episode / Acute rejection > 1 episode / p value / Odds Ratio / IC 95%
0-2 copies / 49 / 14 / 0.0056 / 5.25 / (1.59-17.28)
≥ 3 copies / 6 / 9
No chronic rejection / Chronic rejection / p value / Odds Ratio / IC 95%
0-2 copies / 84 / 35 / 0.18 / 0.48 / (0.15-1.51)
≥ 3 copies / 20 / 4

Supplementary Table 2. Allelic frequencies of CCL4L polymorphisms (rs4796195 and rs3744595) in control population and different groups of LT patients.

Group / Control / Total transplanted / No acute rejection / Acute rejection / Acute rejection > 1 episode / No chronic rejection / Chronic rejection
Nº of samples / n = 80 / n = 161 / n = 79 / n = 78 / n = 23 / n = 104 / n = 39
CCL4L / Total copies / 149 / 290 / 131 / 153 / 53 / 190 / 69
rs4796195 / L1 frequency
(n) / 83.89%
(125) / 81.03%
(235) / 81.68%
(107) / 81.05%
(124) / 83.02%
(44) / 81.05%
(154) / 79.71%
(55)
L2 frequency
(n) / 16.11%
(24) / 18.97%
(55) / 18.32%
(24) / 18.95%
(29) / 16.98%
(9) / 18.95%
(36) / 20.29%
(14)
rs3744595 / R frequency
(n) / 91.95%
(137) / 92.41%
(268) / 93.89%
(123) / 90.85%
(139) / 88.68%
(47) / 91.05%
(173) / 95.65%
(66)
H frequency
(n) / 8.05%
(12) / 7.59%
(22) / 6.11%
(8) / 9.15%
(14) / 11.32%
(6) / 8.95%
(17) / 4.35%
(3)

Supplementary Table 3. Genotypic frequencies of CCL4L polymorphisms (rs4796195 and rs3744595) in control population and different groups of LT patients.

Group / Control / Total transplanted / No acute rejection / Acute rejection / Acute rejection > 1 episode / No chronic rejection / Chronic rejection
Nº of samples / n = 80 / n = 161 / n = 79 / n = 78 / n = 23 / n = 104 / n = 39
rs4796195 / L1 / 68.75% / 67.70% / 72.15% / 62.82% / 52.17% / 66.35% / 69.23%
L1 / L2 / 25.00% / 25.47% / 20.25% / 30.77% / 39.13% / 26.92% / 23.08%
L2 / 2.50% / 3.11% / 3.80% / 2.56% / 0% / 2.88% / 5.13%
- / - / 3.75% / 3.73% / 3.80% / 3.85% / 8.70% / 3.85% / 2.56%
rs3744595 / R / 81.25% / 84.47% / 86.08% / 82.05% / 73.91% / 82.69% / 89.74%
R / H / 15.00% / 11.80% / 10.13% / 14.10% / 17.39% / 13.46% / 7.69%
H / 0% / 0% / 0% / 0% / 0% / 0% / 0%
- / - / 3.75% / 3.73% / 3.80% / 3.85% / 8.70% / 3.85% / 2.56%

The genotypic frequencies were calculated considering that, in the case of genes with CNV, what is usually called ‘homozygous’ and ‘heterozygous’ are those that present one or the two allelic variants respectively regardless of their copy number.

Supplementary Table 4. Mean number of CCL4L copies in control population and LT patient groups based on the allelic variants (L1 and L2) of rs4796195 SNP.

Genotype / n / Mean of CCL4L copies / p value
Controls / L1 / 55 / 1.78 / 0.0018
L2 presence / 22 / 2.32
Total
Transplanted / L1 / 109 / 1.64 / < 0.0001
L2 presence / 46 / 2,41
No acute
rejection / L1 / 57 / 1.49 / < 0.0001
L2 presence / 19 / 2.42
Acute
rejection / L1 / 49 / 1.86 / 0.0023
L2 presence / 26 / 2.39
Acute rejection
> 1 episode / L1 / 12 / 2.41 / 0.3752
L2 presence / 9 / 2.66
No chronic rejection / L1 / 69 / 1.68 / < 0.0001
L2 presence / 31 / 2.39
Chronic
rejection / L1 / 27 / 1.59 / 0.0024
L2 presence / 11 / 2.36

Individuals were classified according their genotype. (L1): individuals homozygous for CCL4L1 allele regardless of their copy number. (L2): individuals carrying at least one copy of CCL4L2 allele regardless of the presence of L1 allele.

Supplementary Table 5. Characteristics of LT recipients and comparison between those possessing low (0-1) or high (2) number of CCL4L copies.

Characteristic / All Recipients (n=161) / Recipients with
0-1 CCL4L copies (n=56) / Recipients with
≥ 2 CCL4L copies
(n=105)
Sex, n (%)
Males
Females / 105 (65)
56 (35) / 32 (57)
24 (43) / 73 (70)
32 (30)
Age, mean  SD / 44 16 / 45  16 / 43  17
Native Disease, n (%)
COPD
Idiopathic pulmonary fibrosis
Cystic fibrosis / bronchiectasis
Pulmonary hipertensión
Others / 67 (42)
39 (24)
32 (20)
11 (7)
12 (7) / 22 (39)
18 (32)
6 (11)
5 (9)
5 (9) / 45 (43)
21 (20)
26 (25)
6 (6)
7 (6)
Type of transplant operation, n (%)
Single
Double / 29 (18)
132 (82) / 12 (21)
44 (79) / 17 (16)
88 (84)
HLA mismatches, mean / 4.6 / 4.7 / 4.4
PRA > 10%, n (%) / 4 (2.5) / 2 (3.6) / 2 (1.9)

COPD: chronic obstructive pulmonary disease; PRA: panel reactive antibody.

Supplementary Table 6. Primers and probes for rs4796195 and rs3744595 CCL4L SNPs genotyping.

SNP / Genotyping
method / Primers (5’-3’) / Probes
rs4796195 / real-time
PCR / sense
GCAGAGGAAGATGCCTACCAC / AGCGCAGACTTGCTTGCCTCTTTTFluos
antisense
CTGAGTATGGAGGAGATGCGG / LC-Red705TTTGGAATCCGTAGAACAAGG
rs3744595 / real-time
PCR / sense
GGAATGGATACAAGGGACCA / CAGAGGCTGCTGGTCTCATAGTAATCTFluos
antisense
CAGCCAGGGGTTGATACT / LC-Red610CCACAAAGTTGCGAGGAA