Tracking GAD-specific T-cell expansions in Type 1 diabetes by intradermal GAD-Alum challenge
We used intradermal injection of GAD-Alum to recruit GAD-specific T cells to accessible sites in the skin and skin-draining lymph nodes (LNs), sampled by skin suction blisters and ultrasound-guided LN aspiration. Peripheral blood samples obtained before GAD injection were restimulated with GAD in vitro to detect reactive CD4+ T cells. Sequencing was performed using 10xGenomics 5' kits (V1 and V2). Samples from LN and blisters were run individually. PBMC samples were combined in a singles 10x reaction using hashing antibodies. LNs include GEX, TCRS and BCRs. Blisters include GEX and TCRs. PBMC include GEX, TCRs, hashing and a panel of cell surface antibodies.
We used intradermal injection of GAD-Alum to recruit GAD-specific T cells to accessible sites in the skin and skin-draining lymph nodes (LNs), sampled by skin suction blisters and ultrasound-guided LN aspiration. Peripheral blood samples obtained before GAD injection were restimulated with GAD in vitro to detect reactive CD4+ T cells. Sequencing was performed using 10xGenomics 5' kits (V1 and V2). Samples from LN and blisters were run individually. PBMC samples were combined in a singles 10x reaction using hashing antibodies. LNs include GEX, TCRS and BCRs. Blisters include GEX and TCRs. PBMC include GEX, TCRs, hashing and a panel of cell surface antibodies.
European Bioinformatics Institute