Next generation sequencing based assessment of the alloreactive T cell receptor repertoire in kidney transplant patients during rejection: a prospective cohort study

Abstract

Background Kidney transplantation is the optimal treatment in end stage renal disease but the allograft survival is still hampered by immune reactions against the allograft. This process is driven by the recognition of allogenic antigens presented to T-cells and their unique T-cell receptor (TCR) via the major histocompatibility complex (MHC), which triggers a complex immune response potentially leading to graft injury. Although the immune system and kidney transplantation have been studied extensively, the subtlety of alloreactive immune responses has impeded sensitive detection at an early stage. Next generation sequencing of the TCR enables us to monitor alloreactive T-cell populations and might thus allow the detection of early rejection events. Methods/design This is a prospective cohort study designed to sequentially evaluate the alloreactive T cell repertoire after kidney transplantation. The TCR repertoire of patients who developed biopsy confirmed acute T cell mediated rejection (TCMR) will be compared to patients without rejection. To track the alloreactive subsets we will perform a mixed lymphocyte reaction between kidney donor and recipient before transplantation and define the alloreactive TCR repertoire by next generation sequencing of the complementary determining region 3 (CDR3) of the T cell receptor beta chain. After initial clonotype assembly from sequencing reads, TCR repertoire diversity and clonal expansion of T cells of kidney transplant recipients in periphery and kidney biopsy will be analyzed for changes after transplantation, during, prior or after a rejection. The goal of this study is to describe changes of overall T cell repertoire diversity, clonality in kidney transplant recipients, define and track alloreactive T cells in the posttransplant course and decipher patterns of expanded alloreactive T cells in acute cellular rejection to find an alternative monitoring to invasive and delayed diagnostic procedures. Discussion Changes of the T cell repertoire and tracking of alloreactive T cell clones after combined bone marrow and kidney transplant has proven to be of potential use to monitor the donor directed alloresponse. The dynamics of the donor specific T cells in regular kidney transplant recipients in rejection still rests elusive and can give further insights in human alloresponse. Trial registration Clinicaltrials.gov: NCT03422224 , registered February 5th 2018. Electronic supplementary material The online version of this article (10.1186/s12882-019-1541-5) contains supplementary material, which is available to authorized users.

Approaches for Controlling Antibody-Mediated Allograft Rejection Through Targeting B Cells

“…Thus, these techniques may permit inhibition of the differentiation of LLPCs. Further; NGS analysis of antibody reactivity to transplanted grafts can be applied to predict the antibodies that cause AMR (28)(29)(30)(31)(32)(33)(34)(35)(36). This knowledge, combined with the identification of genetic polymorphisms associated with drug sensitivity, will undoubtedly contribute to the development of optimal management strategies.…”

“…NGS determines several million base pairs per run and provides the ability to distinguish different isoforms and allelic expression, which is an advantage over microarray analysis and detects somatic mutations with high accuracy and high specificity, which enables identification of candidate genes that cause disease. As an example of the application of NGS to transplantation, comparison of the complementary determining region 3 (CDR3) of the TCR beta chains expressed in AMR (+) and AMR (−), may lead to the prediction of the development of rejection before patients undergo transplantation ( 31 ). In addition, this technique can be applied to increase our understanding of the diversity of the variable regions of heavy and light chains of BCRs.…”

“…Identifying of the antigen specificity of MBCs circulating in the peripheral blood, as measured using an ELISpot assay or in vitro assay system, will be useful to evaluate the activation of the humoral immune response to donor-specific HLA antigens (22)(23)(24)(25)(26)(27). Furthermore, microarray techniques and next-generation sequencing (NGS) may provide detailed information that will contribute to therapeutic control of AMR, including the identification of antibodies that injure transplants and individual differences in patients' susceptibilities to immunosuppressive agents (28)(29)(30)(31)(32)(33)(34)(35)(36).…”

Prospective Tracking of Donor-Reactive T-Cell Clones in the Circulation and Rejecting Human Kidney Allografts

“…The detailed design of this prospective observational cohort study may be found elsewhere ( 19 ). In brief, we enrolled all incident non-sensitized deceased or live donor kidney transplant recipients at our center between November 1, 2017, and September 30, 2019, with a follow-up for 1 year after transplantation or until December 10, 2019, and medical induction therapy was uniformly basiliximab (anti-CD25 antibody) ( 19 ).…”

Precision medicine in transplantation and hemodialysis

“…In order to estimate the risk of a clinically relevant alloimmune response after transplantation, tracking of alloreactive T cells determined by mixed lymphocyte reaction has been suggested previously [ 15 ]. Although the T-cell receptor repertoire is exceedingly complex with high inter-individual diversity, it is nowadays possible to determine the clonality and diversity by DNA sequencing usually of the complementary determining region 3 of the T-cell receptor beta chain [ 16 ]. Even more complex because of somatic hypermutation is the individual tracking of the B-cell alloimmune repertoire and network [ 17 ].…”