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SRX23828826: GSM8124002: VCOV_s03_P0,γδ scTCR-seq; Homo sapiens; OTHER
4 ILLUMINA (NextSeq 550) runs: 75.8M spots, 12.7G bases, 4.6Gb downloads

External Id: GSM8124002_r1
Submitted by: Humanitas Research Hospital
Study: Expansion of memory Vd2 T cells following SARS-CoV-2 vaccination revealed by temporal single-cell transcriptomics
show Abstracthide Abstract
?d T cells provide rapid cellular immunity against pathogens. Here, we conducted matched single-cell RNA-sequencing and ?d-TCR-sequencing to delineate the molecular changes in ?d T cells during a longitudinal study following mRNA SARS-CoV-2 vaccination. While the first dose of vaccine primes Vd2 T cells, it is the second administration that significantly boosts their immune response. Specifically, the second vaccination uncovers memory features of Vd2 T cells, shaped by the induction of AP-1 family transcription factors and characterized by a convergent central memory signature, clonal expansion, and an enhanced effector potential. This temporally distinct effector response of Vd2 T cells was also confirmed in vitro upon stimulation with SARS-CoV-2 spike-peptides. Indeed, the second challenge triggers a significantly higher production of IFN? by Vd2 T cells. Collectively, our findings suggest that mRNA SARS-CoV-2 vaccination might benefit from the establishment of long-lasting central memory Vd2 T cells to confer protection against SARS-CoV-2 infection. Overall design: Peripheral blood mononuclear cells were isolated from buffy coats of 6 healthy volunteers (3 males and 3 females, age range 25-50 years old) with two doses of mRNA COVID-19 (BNT162b2)–vaccine without previous SARS-CoV-2 infection. PB were longitudinally collecteded for each healthy donor 1 day before the first dose (P0; n=6), 3 days after the first vaccine dose (P1; n=6), 17 days after the first dose (P2; n=5), 3 days after the second dose (P3; n=6), and 3 months following the boost (P4; n=5). PB were analyzed using paired single-cell RNA-seq and ?d TCR-seq technics.
Sample: VCOV_s03_P0,γδ scTCR-seq
SAMN40253114 • SRS20650872 • All experiments • All runs
Organism: Homo sapiens
Library:
Name: GSM8124002
Instrument: NextSeq 550
Strategy: OTHER
Source: TRANSCRIPTOMIC
Selection: other
Layout: PAIRED
Construction protocol: Peripheral blood mononuclear cells (PBMCs) were isolated from buffy coats of HDs (n=6) or from 6 healthy vaccinated subjects through Lympholyte®-H Cell Separation density gradient solution (Cedarlane Laboratories, Burlington, North Carolina, USA) according to the manufacturer's instruction. In vitro experiments were performed on freshly isolated PBMCs, with a viability greater than 95%, while single-cell ones on PBMCs previously frozen in Fetal Bovine Serum (FBS) supplemented with 10% of the cryoprotective Dimethyl Sulfoxide (DMSO). Thawed PBMC samples were evaluated for viability prior to scRNA-seq analysis, and all were ≥95% viable. Cells were resuspended in a volume equivalent to 10,000 target cells for each sample, and were individually loaded onto a Chromium single-cell controller (10X Genomics) to generate single-cell gel beads-in-emulsion (GEMs). Captured cells were then lysed and the released RNA was barcoded through reverse transcription in individual GEMs. Complementary DNAs (cDNA) were generated and split to generate γδ scTCR-seq libraries. To this end, gene-specific primers 25 were used within the 5′ regions of the TRGC and TRDC segments for the enrichment of TCR transcripts (v1.1 chemistry) Complementary DNAs were amplified, and the quality was assessed using an Agilent 4200 Tapestation. The scTCR-seq libraries were sequenced on Illumina NextSeq 550 platform.
Runs: 4 runs, 75.8M spots, 12.7G bases, 4.6Gb
Run# of Spots# of BasesSizePublished
SRR2821662618,891,6703.2G1.1Gb2024-03-04
SRR2821662718,765,0523.2G1.1Gb2024-03-04
SRR2821662819,206,0333.2G1.2Gb2024-03-04
SRR2821662918,945,3783.2G1.2Gb2024-03-04

ID:
32130623

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