single cell transcriptomics and B-cell repertoire data from one vaccinated individual after Boostrix vaccination. single cell transcriptomics and B-cell repertoire data from one vaccinated individual after Boostrix vaccination

The transcriptomics and BCR-rep data was meassured at the single-cell level from the longitudinal samples obtained from an individual following Boostrix vaccination. The donor had no known or suspected exposure to pertussis infection nor vaccination in the past 10 years. EDTA-blood samples for single-cell sequencing were collected at baseline and days 5 (d5), d7, d10 and d14 post-vaccination. At baseline, first 10,000 total CD19+ B cells were sorted, and after adjusting the sorting gate to exclusively sort PB/PCs, another 1,201 CD19+CD38++CD24+CD27+ PB/PCs were sorted. For subsequent visits, all available PB/PCs were sorted (11,051 cells). In total, we sorted 22,252 cells. Nearly 22,000 B cells enriched in PB/PCs from different time-points were processed into single cells in a Chromium Controller (10X Genomics). Reverse transcription PCR and library preparation were carried out under the Chromium Single Cell 3’ v3 protocol (10X Genomics) per manufacturer’s recommendations. After amplification of the cDNA, a 5 ́gene expression library and paired heavy and light chain library were generated from cDNA of the same cell using Chromium Single Cell VDJ reagent kit (scBCR-rep library; v1.1chemistry, 10X Genomics). After library preparation, quality control was performed using a bioanalyzer (Agilent 2100 Bioanalyzer; Agilent Technologies). The libraries were sequenced in the NovaSeq6000 sequencer (Illumina) using the v1.0 sequencing reagent kit (read length: 2 x 150bp). Overall design: Longitudinal examination of five different time points at baseline and post vaccination (baseline, days 5, 7, 10 and 14 post vaccination)

本数据集的转录组学与B细胞受体库（BCR-rep）数据来自1名接种Boostrix疫苗后获取的纵向随访样本，以单细胞水平完成检测。该受试者在过去10年内无已知或疑似百日咳感染暴露史，亦未接种过相关疫苗。用于单细胞测序的乙二胺四乙酸（EDTA）抗凝血样本分别于基线期以及疫苗接种后第5、7、10、14天采集。基线期时，先分选得到总计10000个CD19+ B细胞；随后调整分选门控策略，仅分选成浆细胞与浆细胞（PB/PC），又获得1201个CD19+CD38++CD24+CD27+ PB/PC细胞。后续随访时间点的样本中，我们分选了所有可获取的PB/PC细胞，共计11051个。本研究总计分选得到22252个细胞。我们将来自不同时间点、近22000个富集了PB/PC的B细胞通过Chromium Controller（10X Genomics）处理为单细胞悬液以开展后续实验。按照Chromium Single Cell 3’ v3试剂盒（10X Genomics）的制造商操作指南，完成逆转录PCR（reverse transcription PCR）与文库制备。在cDNA扩增完成后，使用Chromium Single Cell VDJ试剂盒（单细胞BCR库文库，scBCR-rep library；v1.1试剂，10X Genomics）从同一细胞的cDNA中构建5'基因表达文库与配对重链、轻链文库。文库制备完成后，使用Agilent 2100生物分析仪（Agilent 2100 Bioanalyzer；Agilent Technologies）完成质量控制。使用v1.0测序试剂盒（读长：2×150bp）在NovaSeq6000测序仪（Illumina）上完成文库测序。整体实验设计：于基线期及疫苗接种后共5个时间点开展纵向检测，分别为基线期、接种后第5、7、10、14天。