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©The Author(s) 2018. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Transplant. Feb 24, 2018; 8(1): 23-37
Published online Feb 24, 2018. doi: 10.5500/wjt.v8.i1.23
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
Francisco Boix, Santiago Llorente, Jorge Eguía, Gema Gonzalez-Martinez, Rafael Alfaro, Jose A Galián, Jose A Campillo, María Rosa Moya-Quiles, Alfredo Minguela, Jose A Pons, Manuel Muro
Francisco Boix, Jorge Eguía, Gema Gonzalez-Martinez, Rafael Alfaro, Jose A Galián, Jose A Campillo, María Rosa Moya-Quiles, Alfredo Minguela, Manuel Muro, Department of Immunology, Clinical University Hospital Virgen de la Arrixaca-IMIB, Clinical University Hospital ‘Virgen Arrixaca’, Murcia 30120, Spain
Santiago Llorente, Department of Nephrology, Clinical University Hospital ‘Virgen de la Arrixaca-IMIB’, Clinical University Hospital ‘Virgen Arrixaca’, Murcia 30120, Spain
Jose A Pons, Digestive Medicine Service, Clinical University Hospital ‘Virgen de la Arrixaca-IMIB’, Clinical University Hospital ‘Virgen Arrixaca’, Murcia 30120, Spain
Author contributions: Muro M planned the experiments, overviewed the research project and reviewed the manuscript; Eguía J, Gonzalez-Martinez G, Alfaro R and Galián JA performed the experimental worked supervised by Boix F; Boix F, Campillo JA and Moya-Quiles MR analysed the data; Minguela A reviewed the manuscript; Llorente S and Pons JA provided clinical data and reviewed the manuscript; Boix F and Muro M equally participated in the writing of the manuscript.
Supported by Instituto de Salud Carlos III, Spanish Ministry of Economy and Competitiveness, No. PI15/01370; Co-funding of the European Union with European Fund of Regional Development (FEDER) with the principle of “A manner to build Europe”.
Institutional review board statement: The study protocol, standard operating procedures and good manufacturing practice protocols used in this research were approved by the institutional ethical committee.
Informed consent statement: Formal informed consent was obtained from both patients and healthy controls.
Conflict-of-interest statement: Authors have no relevant conflicts of interest to disclose.
Data sharing statement: No additional data are available.
Open-Access: This article is an open-access article which was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution Non Commercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/licenses/by-nc/4.0/
Correspondence to: Francisco Boix, PhD, Doctor, Senior Scientist, Department of Immunology, Clinical University Hospital ́‘Virgen de la Arrixaca-IMIB’, Clinical University Hospital ‘Virgen Arrixaca’, Murcia 30120, Spain.
francisco.boix-giner@nhsbt.nhs.uk
Telephone: +34-968-369599 Fax: +34-968-369678
Received: November 6, 2017
Peer-review started: November 7, 2017
First decision: November 20, 2017
Revised: January 13, 2018
Accepted: February 4, 2018
Article in press: February 5, 2018
Published online: February 24, 2018
Processing time: 109 Days and 22 Hours
AIM
To validate intracellular cytokine production functional assay as means of cell-mediated immunity monitoring of post-transplant patients with opportunistic infection (OI).
METHODS
Intracellular cytokine-producing CD4+ and CD8+ T-cell monitoring was carried out in 30 liver transplant (LTr) and 31 kidney transplant (KTr) recipients from 2010 to 2012. Patients were assessed in our Department of Immunology at the Clinical University ‘Hospital Virgen de la Arrixaca-IMIB’ in Murcia, Spain for one year following transplantation. FACS Canto II flow cytometer was employed to quantify the intracellular production of IL-17, IFNγ and IL-10 cytokines on stimulated CD4+CD69+ and CD8+CD69+ T cells and BD FACS DIVA v.6 software was used to analysed the data. Statistical analysis was carried out using SPSS 22.0.
RESULTS
LTr with OI had significantly lower % of CD8+CD69+IFNγ+ T cells at 60 (7.95 ± 0.77 vs 26.25 ± 2.09, P < 0.001), 90 (7.47 ± 1.05 vs 30.34 ± 3.52, P < 0.001) and 180 (15.31 ± 3.24 vs 24.59 ± 3.28, P = 0.01) d post-transplantation. Higher % of CD4+CD69+IL-10+ as well as CD4+CD69+IL-17+ T cells were yet reported at 30 (14.06 ± 1.65 vs 6.09 ± 0.53, P = 0.0007 and 4.23 ± 0.56 vs 0.81 ± 0.14, P = 0.005; respectively), 60 (11.46 ± 1.42 vs 4.54 ± 0.91, P = 0.001 and 4.21 ± 0.59 vs 1.43 ± 0.42, P = 0.03; respectively) and 90 d (16.85 ± 1.60 vs 4.07 ± 0.63, P < 0.001 and 3.97 ± 0.43 vs 0.96 ± 0.17, P = 0.001). Yet, KTr with OI had significantly lower percentage of CD4+CD69+IFNγ+ at 30 (11.80 ± 1.59 vs 20.64 ± 3.26, P = 0.035), 60 (11.19 ± 1.35 vs 15.85 ± 1.58, P = 0.02), 90 (11.37 ± 1.42 vs 22.99 ± 4.12, P = 0.028) and 180 (13.63 ± 2.21 vs 21.93 ± 3.88, P = 0.008) d post-transplantation as opposed to CD4+CD69+IL-10+ and CD8+CD69+IL-10+ T cells which percentages were higher at 30 (25.21 ± 2.74 vs 8.54 ± 1.64, P < 0.001 and 22.37 ± 1.35 vs 17.18 ± 3.54, P = 0.032; respectively), 90 (16.85 ± 1.60 vs 4.07 ± 0.63, P < 0.001 and 23.06 ± 2.89 vs 10.19 ± 1.98, P = 0.002) and 180 (21.81 ± 1.72 vs 6.07 ± 0.98, P < 0.001 and 19.68 ± 2.27 vs 10.59 ± 3.17, P = 0.016) d post-transplantation. The auROC curve model determined the most accurate cut-off values to stratify LTr and KTr at high risk of OI and Cox Regression model confirmed these biomarkers as the most significant risk factors to opportunistic infection.
CONCLUSION
Post-transplant percentages of T-cell subsets differed significantly amongst infected- and non-infected-LTr and -KTr and yet this imbalance was found to contribute towards a worst clinical outcome.
Core tip: The aim of this research was to validate predictive biomarkers for the occurrence of post-transplant opportunistic infection in both liver and kidney recipients. The imbalance in the percentage of cytokine-producing cultured CD4+CD69+ and CD8+CD69+ T cells was shown to be the most significant recipient risk factor to develop opportunistic infection.
