scRNAseq gene expression and VDJ-TCR analysis of PBMCs from patients with tattoo uveitis and sarcoidosis
To elucidate the complexity of immune responses in the formation of granuloma, we performed the single-cell RNA-sequencing analysis. PBMCs derived from tattoo uveitis patients in both active and inactive stages and sarcoidosis patients were collected to characterize their similarities and difference. PBMCs from age-matched healthy donors were considered as controls. Overall design: Sorted live PBMCs were prepared two patients with active tattoo uveitis, three patients with convalescent tattoo uveitis, three patients with sarcoidosis and three age-match healthy donors. Single-cell gene expression and TCR VDJ libraries were prepared by Chromium Next GEM Single Cell 5' Kit v2 Dual Index and Single Cell Human TCR Amplification Kit (10x Genomics) with the help from Biomolecular Resource Facility at ANU. Libraries were then sequenced on NovaSeq 6000 sequencer (Illumina) with 2 x 50 bp paired-end reads. 10X Cell Ranger (10X Genomics, v6.0.1) was utilized to align the reads to the human genome (GRCh38-2020-A) using default parameters.