GSE191088
Comprehensive comparison of adaptive immune response by inactivated SARS-CoV-2 vaccine between young and old [RNA-seq]
Purpose:Comprehensively compared the adaptive immune response of SARS-CoV-2 inactivated vaccines in young and elderly. Methods:CD8+ T, CD4+ T and B cells were purified from PBMCs with EasySep Human  positive/negative selection . PBMCs were incubated with antibody cocktail and then RapidSpheres, then the magnet was applied and unbound CD8+ T, CD4+ T and B cells were recovered from the supernatant. Briefly, the cells were stained with the corresponding CD8+ T, CD4+ T and B-cell  antibodies for 30 minutes at 4°C in the dark, and the purity of the cells was detected separately by flow cytometry, and all could reach more than 95%. Total RNA was isolated from CD8+ T, CD4+ T and B cells of 3 young and older people using TRIzol Reagent (Invitrogen)  (7-days post second vaccination dose) . RNA purity was checked by the NanoPhotomerer spectrophotometer (IMPLEN), and integrity was assessed using the RNA Nano 6000 Assay Kit of the Bioanalyzer 2100 system (Agilent Technologies). Then cDNA libraries were constructed using 0.1 µg RNA per sample with the NEBNext UltraTM RNA Library Prep Kit for Illumina (NEB) following manufacturer's recommendations and index codes were added to attribute sequences to each sample. The clustering of the index-coded samples was performed on a cBot Cluster Generation System using TruSeq PE Cluster Kit v3-cBot-HS (Illumia). After cluster generation, the library preparations were sequenced on an Illumina Novaseq platform and 250 bp paired-end reads were generated. The result showed the inactivated vaccine is less protective in older adults, who take longer to develop effective antibodies, and the TCR diversity of each epitope specific repertoire decreased in the elderly. In addition, we found inactivated vaccines could stimulate the proliferation of related B cells in the body, thereby reducing the diversity of BCR in the body. Compared with the young, the elderly is less likely to produce antibody related BCR clones and the same is true for TCR diversity. Conclusions: The result showed the inactivated vaccine is less protective in older adults, who take longer to develop effective antibodies, and the TCR diversity of each epitope specific repertoire decreased in the elderly. In addition, we found inactivated vaccines could stimulate the proliferation of related B cells in the body, thereby reducing the diversity of BCR in the body. Compared with the young, the elderly is less likely to produce antibody related BCR clones and the same is true for TCR diversity. Overall design: A total of 36 fastq files from the young and old, pre- and post- vaccine of CD4+ T, CD8+ T and B cells were finally got.