Literature sharing Single-cell TCR sequencing: Analyzing immunohistocologic characteristics of renal cell carcinoma

Preface:

T cells are an important component of the acquired immune response, and their surface receptors (T cell receptors) (TCRs) act to recognize antigen molecules

Basic information about the paper

• Title: Characteristics, dynamic changes, and prognostic significance of TCR repertoire profiling in renal cell carcinoma patients

Renal cell carcinoma (RCC) is a malignant tumor originating in the tubular epithelium, accounting for 80% to 90%

Although RCC is considered a highly immunogenic cancer, immunosuppression often occurs

★ Research methodology

To study the characteristics of TCRB, 87 PBMC samples were first obtained from 45 patients with RCC and 2 patients with benign kidney disease, then RNA was extracted and a TCPB sequencing library

• Diversity of TCRBs was positively correlated with the number of Native T cells (particularly CD4^- and CD8^- naïve T cells

■ High sensitivity—Integrate SMART technology with 5'RACE technology to capture intact V(D)J variable regions of TRA and TLB genes for subsequent NGS analysis

■ Easy to use—Optimized index allows 96 cells to be mixed into 12 libraries, and can be further mixed in a flow-cell lane for sequencing

Experimental well plate layout

The negative/positive control group outside the well plate is used for quality assessment of the sequencing library, and the negative/positive control wells within the plate are used for threshold setting when clonal identification analysis and for the assessment

Single-cell sequencing involves a greater number of libraries, and labeling each single-cell library is critical

Experimental cases

At least one TCR chain was identified at 92% (81/88 wells), 65% (57/88 wells) for α/β clones, and 73% (64/88 wells)

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