Single-cell analysis of Diphtheria, Tetanus, and Pertussis (DTP)-vaccinated repertoire

Diphtheria, tetanus, and pertussis (DTP) are infectious diseases caused by toxin-producing bacteria that can be fatal, especially in children. Despite the availability of combined DTP vaccines for more than 60 years, a comprehensive understanding of how human antibodies respond to DTP vaccination, which helps further improve the vaccine remains elusive. Here, we aimed to characterize toxin-specific antibody repertoires following DTP vaccination. To this end, CD19+CD27+ antigen-experienced B cells obtained from four healthy donors 7-days post vaccination were sorted for toxin-binding and non-toxin-binding B cells, using each of the DTP recombinant toxins (DT, TT, and PT) as fluorescent bait. 10X single-cell immune profiling was then performed, followed by in silico antibody sequence clustering. Antigen-experienced B cells (CD19+CD27+) from four healthy donors vaccinated by Diphtheria, Tetanus, and Pertussis (DTP) vaccine booster were sorted using FACS to obtain DT, TT, or PT binding cells, as well as DTP non-binding cells (NEG). Donor information was retained using Cell Hashtag. Samples were analyzed for scRNA-Seq with additional Feature Barcode and VDJ sequencing.