An efficient low cost method for gene transfer to T lymphocytes.

UNLABELLED: Gene transfer to T lymphocytes has historically relied on retro and lentivirus, but recently transposon-based gene transfer is rising as a simpler and straight forward approach to achieve stable transgene expression. Transfer of expression cassettes to T lymphocytes remains challenging, being based mainly on commercial kits. AIMS: We herein report a convenient and affordable method based on in house made buffers, generic cuvettes and utilization of the widely available Lonza nucleofector II device to promote efficient gene transfer to T lymphocytes. RESULTS: This approach renders high transgene expression levels in primary human T lymphocytes (mean 45%, 41-59%), the hard to transfect murine T cells (mean 38%, 36-42% for C57/BL6 strain) and human Jurkat T cell line. Cell viability levels after electroporation allowed further manipulations such as in vitro expansion and Chimeric Antigen Receptor (CAR) mediated gain of function for target cell lysis. CONCLUSIONS: We describe here an efficient general protocol for electroporation based modification of T lymphocytes. By opening access to this protocol, we expect that efficient gene transfer to T lymphocytes, for transient or stable expression, may be achieved by an increased number of laboratories at lower and affordable costs

Self-Compassion Interventions and Psychosocial Outcomes: a Meta-Analysis of RCTs

Objectives Self-compassion is a healthy way of relating to one’s self motivated by a desire to help rather than harm. Novel self-compassion-based interventions have targeted diverse populations and outcomes. This meta-analysis identified randomized controlled trials of self-compassion interventions and measured their effects on psychosocial outcomes. Methods This meta-analysis included a systematic search of six databases and hand-searches of the included study’s reference lists. Twenty-seven randomized controlled trials that examined validated psychosocial measures for self-compassion-based interventions met inclusion criteria. Baseline, post and follow-up data was extracted for the intervention and control groups, and study quality was assessed using the PRISMA checklist. Results Self-compassion interventions led to a significant improvement across 11 diverse psychosocial outcomes compared with controls. Notably, the aggregate effect size Hedge’s g was large for measures of eating behavior (g = 1.76) and rumination (g = 1.37). Effects were moderate for self-compassion (g = 0.75), stress (g = 0.67), depression (g = 0.66), mindfulness (g = 0.62), self-criticism (g = 0.56), and anxiety (g = 0.57) outcomes. Further moderation analyses found that the improvements in depression symptoms continued to increase at follow-up, and self-compassion gains were maintained. Results differed across population type and were stronger for the group over individual delivery methods. Intervention type was too diverse to analyze specific categories, and publication bias may be present. Conclusions This review supports the efficacy of self-compassion-based interventions across a range of outcomes and diverse populations. Future research should consider the mechanisms of change