Activated CD4 T cells in tuberculosis express OX40, a target for host-directed immunotherapy [Human_CSF_T_cell_scRNASeq]

Define genes expressed by CD4 and CD8 T cells FACS sorted from a CSF specimen of a human subject with HIV-associated tuberculous meningitis Cells from the CSF of an individual treated for HIV-associated TB meningitis was obtained from a subject enrolled in the HARVEST (ISRCTN15668391) treatment study. Cells were obtained on either the day of study enrollment, initially stored at 4˚C, then cells were isolated by centrifugation at 300xg for 10 minutes within 6 hours of collection. The CSF cell pellet used for scRNA-Seq was resuspended in freezing medium (FCS with 10% DMSO) prior to cryopreservation at -80˚C using a Mr. Frosty and final storage at -150˚C. Within 6 months of cryopreservation, cells were transported at -50˚C, cryo-recovered, washed with complete RPMI medium, and labeled with a fluorescent viability dye and antibodies for sorting on a MA900 Multi-Application Cell Sorter (Sony Biotechnology) into DMEM with 40% FBS.