Single-Cell RNA sequencing reveals prophylactic Prednisone blunts cellular immunity in hemophilia B gene therapy

Seven hemophilia B gene therapy participants were taken blood for single cell sequencing at four timepoints, and then compared the immune status of T and antigen presentation cells of theses patients according to therapy efficacy and different time points. The results showed that cellular immunity was elevated in patients with FIX:C less then 40IU/dl at week 26, glucocorticoids (GCs) effectively suppressed T cell immunity and GCs inhibited T cell immunity indirectly by effecting on APCs. Examination of single cell mRNA profile in peripheral blood mononuclear cell from seven hemophilia B gene therapy participants at following time points：Day-7(before glucocorticoids treatment), Day 0（before AAV infusion）, Day 7(after AAV infusion) and 2 weeks after glucocorticoids withdrawal(stop GCs) .

本研究纳入7名乙型血友病（hemophilia B）基因治疗受试者，于四个预设时间点采集外周血样本开展单细胞测序，并依据治疗疗效与时间节点，对比分析受试者的T细胞与抗原呈递细胞（antigen presentation cells, APCs）免疫状态。研究结果显示：治疗第26周时，凝血因子IX活性（FIX:C）低于40IU/dl的受试者细胞免疫水平显著升高；糖皮质激素（glucocorticoids, GCs）可有效抑制T细胞免疫，且可通过作用于APCs间接调控T细胞免疫功能。本研究针对上述7名受试者的外周血单个核细胞（peripheral blood mononuclear cell, PBMC）进行单细胞mRNA谱分析，具体采样时间点包括：糖皮质激素治疗前第7天（Day-7）、腺相关病毒（adeno-associated virus, AAV）载体输注前第0天（Day 0）、AAV载体输注后第7天（Day 7），以及糖皮质激素停药后2周。