CITE-Seq analysis of immune populations in paired human epicardial adipose tissue and right atrial appendage tissue samples

The primary aim of the study was to compare the immune repertoire of human epicardial adipose tissue with the underlying myocardium with a focus on T cell subsets and specifically tissue-resident memory T cell populations. Overall design: Two paired human epicardial adipose tissue and right atrial appendage tissue specimens were obtained during open chest cardiac surgery. The tissue specimens were enzymatically digested to isolate the stromal vascular fraction. CD45+ immune fraction was then isolated through PE bead enrichment using Nanobeads. Cells were stained with CITE-Seq antibodies (antibody-derived tags) and further CD45+ enrichment was performed using fluorescence-activated cell sorting (FACS). The samples were maintained at a concentration of ~1000 cells/µL and processed as per the 10x Genomics CITE-Seq workflow with TotalSeq(TM)-C for cell surface protein and antigen specificity.. ADT Description Barcode Sequence TotalSeq™-C0138 anti-human CD5 Antibody CATTAACGGGATGCC TotalSeq™-C0358 anti-human CD163 Antibody GCTTCTCCTTCCTTA TotalSeq™-C0160 anti-human CD1c Antibody GAGCTACTTCACTCG TotalSeq™-C0049 anti-human CD3 Antibody TATCCCTTGGGATGG TotalSeq™-C0072 anti-human CD4 Antibody TGTTCCCGCTCAACT TotalSeq™-C0080 anti-human CD8a Antibody GCTGCGCTTTCCATT TotalSeq™-C0087 anti-human CD45RO Antibody CTCCGAATCATGTTG TotalSeq™-C0148 anti-human CD197 (CCR7) Antibody AGTTCAGTCAACCGA TotalSeq™-C0146 anti-human CD69 Antibody GTCTCTTGGCTTAAA TotalSeq™-C0088 anti-human CD279 (PD-1) Antibody ACAGCGCCGTATTTA TotalSeq™-C1046 anti-human CD88 (C5aR) Antibody GCCGCATGAGAAACA