Post Operative Monitoring of Donor Specific T Cells Response as a Prognostic Factor in Organ Transplantation by the Assay Using the Indirect Presentation of Autologous Dendritic Cells

Abstract

In spite of current immunosuppressive regimen, chronic allograft nephropathy remains the crucial clinical barrier for long term survival. The aim of present study is to elucidate whether the study regarding indirect allorecognition predicts the well functioning or ongoing rejection kidney allograft. As an assay for indirect T cell recognition in vitro, we performed indirect MLR in 10 kidney transplantation patients. Five patients are with a well preserved graft function at post operative 8-11 years and others with a poor functioning graft at 2-7 years. Because T cell recognition of alloantigens presented by recipient antigen presenting cells, namely indirect pathway, plays a critical role in the development and progression of chronic allograft nephropathy, we generate the alloantigen presenting autologous dendritic cells (DC) from recipient peripheral blood monocytes. We cultured monocytes with GM-CSF plus IL-4 for 6 days and induced their maturation with TNF-α and PGE2 for additional 2 days in the presence donor cell lysate as an alloantigen. The T cell mediated responses were characterized by the indirect MLR using autologous DCs as well as the direct MLR using donor PBMC. Cytokines or chemokines released from indirect and direct MLR, and cytokines from patient serum were measured by Luminex assay. Patient with poor functioning graft had elevated proinflammatory mediators such as MMP-1 and IP-10 in the sera (p< 0.05) but other cytokines were not significantly different between the 2 groups. In the cytokine assay for indirect MLR, all the patients with functioning graft had significantly elevated cytokine secretion such as IL-4, IL-5 and IL-10 compared with poor functioning group. However, IL-12(p70), IL-23 and IL-1β were increased in the poor functioning group. Interestingly, no significant difference was found in the cytokine levels in direct MLR assay, although IL-10 and IL-5 was increased in some well functioning patients (3 out of 5) compared with poor functioning group. From these results, we can conclude that the assay of cytokine released in indirect MLR might be a favorable predictor for long-term function in renal allograft patient in addition to as a non invasive method. Wee need further study to confirm this result in more patient pool.