Targeted immunosuppression enhances repeated gene delivery

Immune system responses against adeno-associated virus (AAV) vectors are potentiated after the first administration, which has prevented the clinical use of repeated administration of AAV-based gene therapies. Here, we quantify the contributions of multiple immune system components towards AAV response in mice. We identify B-cell-mediated immunity, specifically the generation of IgM antibodies, as a critical component preventing vector re-administration. Single-cell 5' RNA-seq (scRNA-seq) of spleens from C57BL/6 mice that recieved a single-, double- or control AAV injection. All samples are pooled from 3 independent mice in the same condition and processed using 10x Chromium NextGEM Single Cell Immune Profiling Solution 5’v2 according to the manufacturer's instructions.