Mixed Lymphocyte Cultures Can Predict TCR Vβ Repertoires of T Cells Infiltrating Kidney Transplants during Acute Rejection Episodes

Abstract

Alloreactive T cell populations can show skewing of T-cell antigen receptor (TCR) Vbeta gene usage. The aims of the experiments were to compare in vivo and in vitro T cell alloresponses against donor alloantigens for TCR Vbeta gene usage. T-cell cultures from renal biopsies taken during acute rejection and pretransplant mixed lymphocyte cultures (MLC) were established from five renal transplant patients. TCR Vbeta gene usage, assessed with Vbeta family specific antibodies, showed that up to five different Vbeta families were significantly expanded. In four of five cases, there was close concordance between Vbeta families expanded from the biopsy and in MLC. T-cell clones from one renal biopsy were specific for the mismatched donor alloantigen and showed similar TCR Vbeta gene usage to the original T-cell line. The results show very similar patterns of TCR Vbeta gene usage in alloreactive T cells generated ex vivo or in vitro.

Longitudinal Analysis of the T-cell Receptor Repertoire in Graft-infiltrating Lymphocytes Following Hand Transplantation

“…Prior characterization of TCR repertoires by cruder spectratyping-based approaches demonstrated that the expansion of alloreactive GIL clones was not mirrored in the peripheral blood, resulting in a clear distinction in TCR repertoires of oligoclonal tissue-derived and more broadly distributed blood-derived T-lymphocyte populations. 13,15,16,23-25,32-35 Our NGS-based TCR repertoire analysis confirms this phenomenon, in agreement with an earlier study. 18 In greater detail, we now show further that tissue- and blood-derived TCR repertoires have distinct clonal compositions that remain consistent over time.…”

“…Alloreactive GILs are significantly enriched in graft tissues compared with peripheral blood, 15 even in the absence of histopathologically defined rejection. 18 Therefore, these T lymphocytes would be highly represented among the TCR clones in the tissue-derived repertoires and minimally among blood repertoires.…”

“…10-12 Graft-infiltrating lymphocytes (GILs) can be detected throughout the period of histopathologically defined rejection. 13-16 The population of T lymphocytes that comprise alloreactive GILs can be established through mixed lymphocyte reactions involving ex vivo stimulation of recipient peripheral blood mononuclear cells (PBMCs) by donor MHC-presenting cells. 15,17-19…”

“…13-16 The population of T lymphocytes that comprise alloreactive GILs can be established through mixed lymphocyte reactions involving ex vivo stimulation of recipient peripheral blood mononuclear cells (PBMCs) by donor MHC-presenting cells. 15,17-19…”

Clonal CD8+ T Cell Persistence and Variable Gene Usage Bias in a Human Transplanted Hand

“…Understanding and predicting BV usage bias in graft rejection could be clinically useful. Prediction of BV usage useful for therapeutic specificity by targeting only those BV families involved in rejection for deletion [ 14 , 30 ], rather than nonspecifically via agents such as OKT3. In genetically controlled animal transplant models, BV usage of alloreactive T cells is predictable [ 29 , 31 , 32 ] and stable over time [ 22 , 33 ].…”