Figure S1, SDC: Distribution of patients regarding the 2 biomarkers.
1a. All patients.
1b. Excluding the IR-kSORT scores.
1c. Mean donor-specific (d-s) IFN-γ producing T-cell frequencies in HR kSORT, LR kSORT and IR kSORT kidney transplant patients at 6 months after transplantation.
Figure S2, SDC: Six-month kSORT and IFN-γ ELISPOT data and histological lesions at 6-month protocol biopsies.
Figure S3, SDC: 6-months kSORT and ELISPOT data and histological lesions at 6-months protocol biopsies excluding patients
with previous clinical BPAR.
3a. 6-months kSORT and sc-AR.
3b. 6-months IFN-γ ELISPOT and sc-AR.
3c. Combination of the kSORT and IFN-γ ELISPOT assays and sc-AR.
Figure S4, SDC. T-cell depletion (rATG) as induction therapy did not affect the
association of the 2 biomarkers with the presence of sc-AR.
4a. 6-month kSORT and sc-AR in rATG treated patients or not.
4b. 6-month ELISPOT and sc-AR in rATG treated patients or not.
Figure S5, SDC. Kidney allograft function (serum creatinine and proteinuria) progression at 6, 12 and 24 months in E+/HR kSORT versus E-/LR kSORT patients and sc-AR versus STA patients.
5a. Serum creatinine (sCreat) levels at 6, 12 and 24 months in E+/HR kSORT
vs E-/LR kSORT patients and sc-AR vs STA patients.
5b. Proteinuria (mg/dL) levels at 6, 12 and 24 months in E+/HR kSORT vs E- /LR kSORT and sc-AR vs STA patients.
Table S1, SDC. Baseline demographics and 6-month biomarkers.
Baseline Demographics (n=75)
6-mo kSORT 6-mo d-s IFN-γ ELISPOT
High-Risk (n=15; 20%)
Indeterm-Risk (n=10; 13.3%)
Low-Risk (n=50; 66.7%)
p-value POSITIVE (n=31; 41.3%)
NEGATIVE (n=44; 58.7%)
p-value
Donor Age (years) 50±15.1 57±9.2 52±16.2 0.52 56.1±13.6 49.4±15.8 0.06
Recipient Age (years) 53.6±12.9 54.6±9.2 47.9±13.9 0.17 52.9±13.4 48.2±13.2 0.19
Recipient Gender (F) (%) 6(40) 4(40) 21(42) 0.99 10(32.3) 21(47.7) 0.18
Caucasian ethnicity (%) 14(93.3) 9(90) 47(94) 0.90 28(90.3) 42(95.5) 0.38 Cause of ESRD (%)
- Unknown - Glomerular - Interstitial - Vascular - Diabetes - APKD
6(40) 3(20) 0(0) 4(26.7)
0(0) 2(13.3)
3(30) 3(30) 0(0) 1(10) 3(30) 1(10)
11(22) 12(24) 11(22) 5(10)
2(4) 9(18)
0.32
10(32.3) 9(29) 5(16.1)
3(9.7) 1(3.2) 3(9.7)
10(22.7) 8(18.2) 6(13.6) 7(15.9) 4(9.1) 9(20.5)
0.48
Donor type (Deceased) (%) 12(80) 8(80) 38(76) 0.93 22(71) 36(81.8) 0.27
Number of transplants 1.1±0.3 1.1±0.3 1.1±0.5 0.91 1.1±0.3 1.1±0.5 0.47
Dialysis duration (months) 31.8±17.7 44.6±21.8 36.6±44.4 0.73 49.6±52.6 30.5±24.6 0.06 Number HLA mismatch 3.86±1.03 3.33±1.41 3.94±1.11 0.34 4.03±1.09 3.73±1.17 0.26 Pre-TX HLA Ab (%)
- DSA
6(40) 1(6.7)
2(22.2) 0(0)
7(14) 1(2)
0.08 0.53
7(23.3) 1(3.2)
8(18.2) 1(2.3)
0.58 0.78 Induction therapy (yes) (%)
-T-cell depletion (rATG) (%)
13(86.7) 6(40)
8(80) 0(0)
47(94) 19(38)
0.32 0.06
26(52) 6(19.4)
42(95.5) 19(43.2)
0.09 0.03
CNI-based IS (%) 12(80) 10(100) 48(96) 0.16 29(93.5) 41(93.2) 0.22
DGF (%) 10(18.9) 3(13.3) 14(63.6 0.90 9(40.9) 13(59.1) 0.90
CMV infection (%) 1(6.7) 1(10) 3(6) 0.88 1(3.2) 4(9.1) 0.33
BPAR (%) 6(40) 2(20) 5(10) 0.03 7(22.5) 6(13.6) 0.31
Kidney Graft Loss (%) 1(6.7) 0(0) 1(2) 0.10 1(3.2) 1(2.3) 0.20
Abbreviations: kSORT: Kidney Solid Organ response test; ELISPOT: Enzyme-linked immunosorbent spot assay; mo: months; d-sp:
donor-specific; Indeterm: indeterminate; ESRD: end-stage renal disease; APKD: Autosomal Polycystic kidney disease; Pre-TX:
pretransplant; CNI: calcineurin-inhibitor; DGF: delayed graft function; CMV: cytomegalovirus infection; BPAR: biopsy-proven acute rejection.
Table S2, SDC: Predictive performance of kSORT and ELISPOT assays for predicting sc-AR, sc-TCMR and sc-ABMR in all patients and excluding patients with previous BPAR
ROC curve analysis
AUC 95%CI AUC p value Sensitivity Specificity PPV NPV All patients of the study
kSORT
- sc-AR 0.732 0.604-0.859 0.002 70 97.7 93.3 88
- sc-TCMR 0.700 0.554-0.846 0.014 60 97.7 90 88
- sc-ABMR 0.840 0.742-0.938 0.013 100 97.7 83.3 100
ELISPOT
- sc-AR 0.690 0.556-0.824 0.010 68.2 69.8 48.4 84.1
- sc-TCMR 0.761 0.632-0.890 0.001 82.4 69.8 46.7 92.5
- sc-ABMR 0.449 0.195-0.703 0.130 20 69.8 5.8 90.2
kSORT and ELISPOT
- sc-AR 0.916 0.840-0.992 <0.001
- sc-TCMR 0.889 0.794-0.984 <0.001 - sc-ABMR 0.989 0.963-1.000 <0.001 Excluding patients with BPAR
kSORT
- sc-AR 0.737 0.586-0.889 0.007 69.2 100 100 91.1
- sc-TCMR 0.691 0.505-0.876 0.060 55.5 100 100 91.1
- sc-ABMR 0.854 0.754-0.954 0.020 100 100 100 100 ELISPOT
- sc-AR 0.711 0.555-0.868 0.017 71.4 70.8 41.7 89.5
- sc-TCMR 0.804 0.667-0.942 0.003 90 70.8 39.1 97.1
- sc-ABMR 0.479 0.187-0.771 0.891 25 70.8 6.6 91.9
kSORT and ELISPOT
- sc-AR 0.947 0.892-1.000 <0.001
- sc-TCMR 0.920 0.845-0.994 <0.001
- sc-ABMR 1.000 1.000-1.000 <0.001
Table S3, SDC. Flow-chart of the value of combining the kSORT and ELISPOT assays to assess the risk of subclinical allograft damage.
HR-kSORT / DSA + High Risk of sc-ABMR HR-kSORT / ELISPOT + High Risk of sc-TCMR
LR-kSORT /ELISPOT- STABLE
