ISSN Print 2500–1094
ISSN Online 2542–1204
BIOMEDICAL JOURNAL OF PIROGOV UNIVERSITY (MOSCOW, RUSSIA)
ORIGINAL RESEARCH
Long-term effect of high cyclophosphamide doses on the repertoire of T-cell receptors of peripheral blood T-lymphocytes in patients with autoimmune vasculitis
About authors
1 Group of structural Organization of T-cell Immunity, Department of Adaptive Immunity Genomics, Shemyakin and Ovchinnikov Institute of Bioorganic Chemistry, Moscow
2 I. M. Sechenov First Moscow State Medical University, Moscow, Russia
Correspondence should be addressed: Olga V. Britanova
ul. Miklukho-Maklaya, d. 16/10, Moscow, Russia, 117997; moc.liamg@natirblo
About paper
Funding: this work was supported by the Russian Science Foundation (Grant No. 16-15-00149). Equipment used in the study was provided by the shared facility of the Institute of Bioorganic Chemistry (the facility is supported by the Ministry of Education and Science of the Russian Federation, project ID RFMEFI62117X0018).
All authors' contribution to this work is equal: selection and analysis of literature, research planning, data collection, analysis, and interpretation, drafting of a manuscript, editing.
Received: 2017-10-04
Accepted: 2017-10-25
Published online: 2018-01-14
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Fig. 1. Preparation of TCR cDNA libraries. The switch adaptor carries a unique sequence (shown as a barcode in the picture) which allows accurate error correction at the stage of data analysis. The amplified fragment carries TCR beta chain CDR3 responsible for receptor diversity
Fig. 2. The proportion of naive T-cells among the peripheral blood CD3+ lymphocytes and the diversity of TCR beta chain repertoires in patients with autoimmune vasculitides who previously received high doses of cyclophosphamide (blue dots) and healthy individuals (orange dots). (А) The proportion of naive T-cells among the total CD3+ lymphocytes plotted against participants’ age. (B) The diversity of TCR estimated with Chao1. The major contribution is made by low-frequency clones, i. e. TCRs of naive lymphocytes. (C) The diversity of CDR3 sequence variants per random 25,000 TCR beta chain cDNA molecules. (D) The diversity of TCR beta chains estimated with the Shannon–Wiener index used to measure the evenness of clonotype distribution in the TCR repertoire. (E) The proportion of naive T-cells among total CD3+ lymphocytes, accounting for Chao1. (F) The number of public clonotypes in clone sets depending on donor’s age. Fig. B–E are based on the analysis of 25,000 randomly selected TCR beta chain cDNA molecules
