RNA-sequencing of Influenza A.Cali09 specific CD4+ T cells from healthy UK adults

Bulk RNA sequencing of sorted A.Cali09-specific CD4+CD45RA- cells from before and seven days after trivalent seasonal influenza vaccination in healthy UK adults of 18-36 years old and over 65 years old. Samples from before and after vaccination were stained with HLADR0701 or HLADR1101 tetramers loaded with influenza peptides specific to the A.Cali09 strain and antigen experienced CD4+CD45RA- T cells sorted by flow cytometry from day 0 and day 7 after vaccination. On day 7, cells were sorted as being CXCR5+ and PD1 expressing circulating Tfh cells. Between 40 and 1000 cells were sorted per sample directly into lysis buffer for subsequent sequencing with the SmartseqV4 kit from Takara Clontech.