Synthesis was primed with the BC_R4_short oligonucleotide (5′-GTATCTGGAGTCATTGA-3′), which is specific to both gene variants of the human TRBC segment. ![]() RNA (1.5 μg) was collected per 15-μl reaction volume, with a total of four to six tubes per sample, such that all RNA was used for each blood sample. Interestingly, cross-analysis of individual TCR β repertoires revealed a set >10,000 of the most representative public TCR β clonotypes, whose abundance among the top 100,000 clones correlated with TCR diversity and decreased with aging.įirst-strand cDNA was synthesized using the Mint cDNA synthesis kit (Evrogen, Russia), according to the manufacturer’s protocol ( Supplemental Fig. Remarkably, the oldest group (average age 82 y) was characterized by a higher percentage of naive CD4 + T cells, lower abundance of expanded clones, and increased TCR diversity compared with the previous age group (average age 62 y), suggesting the influence of age selection and association of these three related parameters with longevity. The percentage of naive T cells showed a strong correlation with measured TCR diversity and decreased linearly up to age 70. We demonstrate that TCR β diversity per 10 6 T cells decreases roughly linearly with age, with significant reduction already apparent by age 40. ![]() In this study, we combined high-throughput Illumina sequencing with unique cDNA molecular identifier technology to achieve deep and precisely normalized profiling of TCR β repertoires in 39 healthy donors aged 6–90 y. ![]() The decrease of TCR diversity with aging has never been studied by direct methods.
0 Comments
Leave a Reply. |
AuthorWrite something about yourself. No need to be fancy, just an overview. ArchivesCategories |