In a forthcoming trial we use mixed lymphocyte reactions to select alloreactice T cell clones to generate a unique fingerprint of the donor-reactive T cell repertoire prior to transplantation. Subtype identification is needed to determine function of the T cells. Clonality and diversity of T cells in biopsies with acute rejection were heterogeneous. Relative abundancies of T cell clonotypes from two patients with acute rejection can be seen in Figure 1: Patient A has a higher clonalization of the repertoire than patient B.Ĭonclusion: We demonstrated that tissue-resident T cell receptor repertoire can be characterized in samples from routine needle biopsies. No significant differences in T cell clonality and diversity were found between acute rejection and no rejection. Results: The single most abundant T cell clonotypes in the kidney accounted for 0.28% to 38.9% of all T cell cells, respectively. Data analysis was done in several steps: first, sequence alignment of the rearranged V(D)J region of the TRB loci by IMGT/HighV-QUEST second, we used our IMEX software for estimating the clonality and the diversity of the aligned. Amplicon sequencing was done by bidirectional Next Generation Sequencing using Illumina Miseq. The rearranged T cell receptor beta (TRB) loci from T cells were amplified by multiplex PCRs based on Biomed-2 primer sets using gDNA as a template. 10 patients had an acute T cell mediated rejection and 10 had no rejection. Methods: We included biopsy samples from 20 patients who were biopsied in the first month after transplantation. We tested the feasibility of TCR repertoire analysis in tissue samples from routine cylinder biopsies of renal allografts. Introduction: Diversity and clonality of the T cell repertoire of circulating and tissue-resident T-cells can be assessed by sequencing of recombined T cell receptor (TCR) genes.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |