In vitro intracellular IFNγ, IL-17 and IL-10 producing T cells correlates with the occurrence of post-transplant opportunistic infection in liver and kidney recipients

Table 1 List of type of opportunistic microorganisms that infected liver and kidney recipients during the post-transplant period n (%)

Type of opportunistic microorganism	Liver transplant recipients (n = 30)	Kidney transplant recipients (n = 31)
Presence of overall opportunistic infection (Yes/No)	18 (60)/12 (40)	19 (61.3)/12 (38.7)
Presence of bacterial infection (Yes/No)	12 (66.7)/6 (33.3)	17 (84.2)/3 (15.8)
Staphylococcus hominis	2 (16.7)/10 (83.3)	1 (5.8)/15 (94.2)
Staphylococcus epidermidis	5 (41.7)/7 (58.3)	5 (29.4)/11 (70.6)
Staphylococcus haemolyticus	2 (16.7)/10 (83.3)	2 (11.8)/14 (88.2)
Enterococcus faecalis	1 (8.3)/11 (91.7)	3 (17.6)/13 (82.4)
Enterococcus faecium	0	3 (17.6)/13 (82.4)
Clostridium difficile	2 (16.7)/10 (83.3)	3 (17.6)/13 (82.4)
Proteus mirabilis	1 (8.3)/11 (91.7)	4 (23.5)/12 (76.5)
Pseudomonas aeruginosa	2 (16.7)/10 (83.3)	5 (29.4)/11 (70.6)
Serratia marcescens	1 (8.3)/11 (91.7)	0
Escherichia coli	2 (16.7)/10 (83.3)	13 (76.5)/3 (23.5)
Treponema pallidum	1 (8.3)/11 (91.7)	0
Enterobacter aerogenes	2 (16.7)/10 (83.3)	1 (5.8)/15 (94.2)
Enterobacter cloacae	0	3 (17.6)/13 (82.4)
Streptococcus sp.	1 (8.3)/11 (91.7)	0
Citrobacter koseri	0	2 (11.8)/14 (88.2)
Morganella morganii	0	1 (5.8)/15 (94.2)
Klebsiella oxytoca	0	2 (11.8)/14 (88.2)
Klebsiella pneumoniae oxytoca	0	1 (5.8)/15 (94.2)
Hafnia alvei	0	1 (5.8)/15 (94.2)
Salmonella typhi	0	2 (11.8)/14 (88.2)
Presence of yeast infection (Yes/No)	4 (22.2)/14 (77.8)	3 (15.8)/16 (84.2)
Candida albicans	4 (100)/0	3 (100)/0
Presence of viral infection (Yes/No)	17 (94.4)/1 (5.6)	17 (89.5)/2 (10.5)
Cytomegalovirus	11 (64.7)/6 (35.3)	12 (70.6)/5 (29.4)
BK virus	3 (17.6)/14 (82.4)	6 (35.3)/11 (64.7)
Epstein-Barr virus	2 (11.8)/15 (88.2)	3 (17.6)/14 (82.4)
Varicella-Zoster virus	2 (11.8)/15 (88.2)	0
Presence of parasitic infection (Yes/No)	3 (16.7)/15 (83.3)	3 (15.8)/16 (84.2)
Toxoplasma gondii	3 (100)/0	2 (66.7)/1 (33.3)
Strongyloides stercoralis	0	1 (33.3)/2 (66.7)

Table 2 Patient clinical and demographic characteristics

	Liver recipients (n = 30)			Kidney recipients (n = 31)
	NoINF (n = 12)	INF (n = 18)	P	NoINF (n = 12)	INF (n = 19)	P
Donor age (yr)	60.75 ± 3.32	58.93 ± 4.71	0.905	51.21 ± 3.171	55.67 ± 3.831	0.0171
Recipient age (yr)	51.25 ± 2.87	53.79 ± 2.37	0.282	51.58 ± 3.25	51.42 ± 2.81	0.351
Recipient gender (M/F), n (%)	14 (87.5)/2 (12.5)	9 (64.3)/5 (35.7)	0.669	10 (41.7)/2 (28.6)	14 (58.3)/5 (71.4)	0.087
Total lymphocyte (%)	15.38 ± 3.63	11.39 ± 2.08	0.397	16.09 ± 2.85	11.66 ± 2.33	0.768
Total lymphocyte (cells/mm3)	813.34 ± 163.70	750.02 ± 191.84	0.711	1121.67 ± 173.35	1073.16 ± 235.68	0.197
Total leukocyte (× 109/L)	6.29 ± 0.55	7.09 ± 1.10	0.652	8.36 ± 1.151	10.98 ± 1.331	0.0061
SGOT (U/L)	187.44 ± 97.28	117.16 ± 18.09	0.738	27.66 ± 4.61	19.17 ± 1.09	0.669
SGPT (U/L)	161.69 ± 69.20	135.56 ± 14.77	0.891	44.51 ± 9.81	27.06 ± 1.98	0.762
SALP (U/L)	177.47 ± 14.72	186.62 ± 14.40	0.847	106.84 ± 8.231	85.36 ± 2.741	0.0081
SGGT (U/L)	197.27 ± 23.651	351.28 ± 42.441	0.0051	109.97 ± 25.34	64.36 ± 11.74	0.074
Glomerular filtration (mL/min)	82.82 ± 7.36	81.28 ± 6.27	0.571	45.87 ± 2.901	72.42 ± 2.521	0.0191
Serum creatinine	0.93 ± 0.051	1.08 ± 0.061	0.0271	6.22 ± 0.60	5.83 ± 0.51	0.251
Induction therapy (thymoglobulin/basiliximab)	0(0 )/0(0)	0(0)/1(3.3)		1(8.3)/11(91.7)	4(21.1)/12(63.2)	0.161
Post-transplant therapy (TRL/TRL + MMF), n (%)	7(58.3)/5(41.7)	10(55.6)/8(44.4)	0.880	0/0/12(100)	1(5.3)/2(10.5)/16(84.2)	0.350
Maintenance therapy (TRL/TRL + MMF), n (%)	7(58.3)/5(41.7)	13(72.2)/5(27.8)	0.461	0/0/12(100)	2(10.5)/2(10.5)/15(78.9)	0.235
TRL dose (mg/d)	7.92 ± 0.281	6.67 ± 0.391	< 0.0011	13.08 ± 1.51	12.35 ± 1.37	0.179
MMF dose (mg/d)	2062.50 ± 73.451	1848.57 ± 79.001	0.0341	1620 ± 164.701	1917.89 ± 58.871	< 0.0011
Cmin TRL (ng/mL)	10.46 ± 0.69	9.61 ± 0.47	0.445	10.54 ± 1.981	6.53 ± 1.971	0.0121
Cmin MMF (μg/mL)	2.91 ± 0.601	0.97 ± 0.291	0.0161	1.07 ± 0.18	3.80 ± 1.20	0.071

¹Statistical significance. NoINF: Free-opportunistic infection study group; INF: Opportunistic infection study group; SGOT: Serum glutamic oxaloacetic transaminase; SGPT: Serum glutamic pyruvic transaminase; SALP: Serum alkaline phosphatase; SGGT: Serum Gamma-Glutamyl Transpeptidase; TRL: Tacrolimus; MMF: Mycophenolic acid.

Table 3 Post-transplant cut-off, area under curve, sensitivity and specificity values for the intracellular cytokine production capacity in liver transplant recipients and kidney transplant recipients as surrogate predictive biomarkers of post-transplant opportunistic infection

Biomarker	Cut-off	AUC (95%CI)	Sensitivity (95%CI)	Specificity (95%CI)
Liver transplant
% CD8+CD69+ IFNγ+ T Lymphocytes	14.95	0.897 (0.834-0.960)	85.71 (75.29-92.93)	88.37 (74.92-96.11)
% CD4+CD69+IL-17+ T Lymphocytes	2.19	0.840 (0.761-0.919)	80.56 (69.53-88.94)	81.25 (67.37-91.05)
% CD4+CD69+IL-10+ T Lymphocytes	9.35	0.902 (0.834-0.969)	78.12 (66.03-87.49)	94.87 (82.68-99.37)
Kidney transplant
% CD4+CD69+ IFNγ+ T Lymphocytes	13.83	0.750 (0.657-0.843)	72.37 (60.91-82.01)	75 (60.40-86.36)
% CD4+CD69+IL-10+ T Lymphocytes	13.95	0.856 (0.792-0.919)	71.05 (59.51-80.89)	83.33 (69.78-2.52)
% CD8+CD69+IL-10+ T Lymphocytes	11.15	0.734 (0.638-0.831)	89.33 (80.06-95.28)	52.17 (36.95-67.11)

AUC: Area under curve.

Table 4 Univariate and multivariate Cox regression model for overall opportunistic infection between 1st and 6th month post-transplantation

Predictive factors for OI	Univariate analysis			Multivariate analysis
	HR	95%CI	P-value	HR	95%CI	P-value
Liver transplant (n = 30)
% CD8+CD69+IFNγ+ T lymphocytes	22.56	3.01-169.56	0.002	21.12	2.80-159.31	0.003
% CD4+CD69+IL-10+ T lymphocytes	3.77	1.46-9.75	0.006	2.84	1.09-7.35	0.032
% CD4+CD69+IL-17+ T lymphocytes	2.43	0.93-6.35	0.07	1.37	0.45-4.16	0.584
Total leukocyte	1.08	0.98-1.18	0.111	0.95	0.78-1.17	0.666
Total lymphocytes (%)	0.94	0.90-0.99	0.022	0.94	0.89-0.99	0.023
Total lymphocyte (cells/mm3)	1.24	1.18-1.30	0.150	1.00	0.99-1.04	0.261
Donor age	0.98	0.96-1.01	0.221	0.96	0.93-0.99	0.026
Recipient gender	2.40	0.94-6.09	0.067	4.56	1.46-14.20	0.009
TRL dose (mg/d)	0.96	0.87-1.10	0.815	0.82	0.59-1.13	0.230
MMF dose (mg/d)	0.87	0.49-1.52	0.619	0.99	0.99-1.01	0.454
Induction therapy	0.92	0.33-2.51	0.864	3.90	3.71-4.09	0.990
Post-transplant IS	1.89	0.78-4.59	0.157	1.98	0.64-6.15	0.238
Acute rejection	1.51	0.63-3.62	0.362	3.95	0.75-20.93	0.107
SALP (U/L)	1.02	0.92-1.17	0.078	1.00	0.99-1.05	0.706
SGGT (U/L)	1.19	1.13-1.25	0.035	1.01	0.91-1.03	0.333
Kindey transplant (n = 31)
% CD4+CD69+IFNγ+ T lymphocytes	2.36	1.28-4.33	0.006	3.29	1.71-6.35	< 0.001
% CD4+CD69+IL-10+ T lymphocytes	3.49	1.68-7.30	0.001	4.76	2.05-11.08	0.003
% CD8+CD69+IL-10+ T lymphocytes	4.17	1.49-11.68	0.007	2.52	0.74-8.57	0.139
Recipient gender	1.91	1.01-3.61	0.046	1.15	0.57-2.32	0.705
TRL dose (mg/d)	0.95	0.89-1.01	0.112	0.92	0.85-0.99	0.037
MMF dose (mg/d)	1.18	1.12-1.24	0.035	1.01	1.00-1.03	0.053
Induction therapy	1.32	0.91-1.91	0.143	1.50	0.96-2.35	0.076
Post-transplant IS	0.94	0.67-1.32	0.723	0.63	0.19-2.01	0.434
Acute rejection	1.34	0.64-2.81	0.439	1.44	0.59-3.48	0.417
Serum creatinine	1.02	1.00-1.03	0.014	1.02	1.00-1.04	0.013
SALP (U/L)	0.98	0.97-0.99	0.016	0.98	0.97-1.01	0.074
SGOT (U/L)	0.98	0.95-1.01	0.242	0.98	0.96-1.01	0.195

IS: Post-transplant immunosuppressive therapy; HR: Hazard ratio.