Bastien Jean-Philippe, Fekete Natalie, Beland Ariane V, Lachambre Marie-Paule, Laforte Veronique, Juncker David, Dave Vibhuti, Roy Denis-Claude, Hoesli Corinne A
Hematology-Oncology and Cell Therapy Institute, Hopital Maisonneuve-Rosemont Research Center, Montreal, Québec, Canada.
Department of Chemical Engineering, McGill University, Montreal, Québec, Canada.
J Transl Med. 2020 Oct 9;18(1):383. doi: 10.1186/s12967-020-02543-1.
A major obstacle to anti-viral and -tumor cell vaccination and T cell immunotherapy is the ability to produce dendritic cells (DCs) in a suitable clinical setting. It is imperative to develop closed cell culture systems to accelerate the translation of promising DC-based cell therapy products to the clinic. The objective of this study was to investigate whether viral antigen-loaded monocyte-derived DCs (Mo-DCs) capable of eliciting specific T cell activation can be manufactured in fluorinated ethylene propylene (FEP) bags.
Mo-DCs were generated through a protocol applying cytokine cocktails combined with lipopolysaccharide or with a CMV viral peptide antigen in conventional tissue culture polystyrene (TCPS) or FEP culture vessels. Research-scale (< 10 mL) FEP bags were implemented to increase R&D throughput. DC surface marker profiles, cytokine production, and ability to activate antigen-specific cytotoxic T cells were characterized.
Monocyte differentiation into Mo-DCs led to the loss of CD14 expression with concomitant upregulation of CD80, CD83 and CD86. Significantly increased levels of IL-10 and IL-12 were observed after maturation on day 9. Antigen-pulsed Mo-DCs activated antigen-responsive CD8 cytotoxic T cells. No significant differences in surface marker expression or tetramer-specific T cell activating potency of Mo-DCs were observed between TCPS and FEP culture vessels.
Our findings demonstrate that viral antigen-loaded Mo-DCs produced in downscaled FEP bags can elicit specific T cell responses. In view of the dire clinical need for closed system DC manufacturing, FEP bags represent an attractive option to accelerate the translation of promising emerging DC-based immunotherapies.
抗病毒和抗肿瘤细胞疫苗接种以及T细胞免疫疗法的一个主要障碍是在合适的临床环境中生产树突状细胞（DC）的能力。开发封闭细胞培养系统以加速有前景的基于DC的细胞治疗产品向临床转化势在必行。本研究的目的是调查在氟化乙烯丙烯（FEP）袋中能否制造出能够引发特异性T细胞活化的负载病毒抗原的单核细胞衍生DC（Mo-DC）。
通过在传统组织培养聚苯乙烯（TCPS）或FEP培养容器中应用细胞因子鸡尾酒与脂多糖或与巨细胞病毒（CMV）病毒肽抗原相结合的方案来生成Mo-DC。采用研究规模（<10 mL）的FEP袋以提高研发通量。对DC表面标志物谱、细胞因子产生以及激活抗原特异性细胞毒性T细胞的能力进行了表征。
单核细胞分化为Mo-DC导致CD14表达丧失，同时CD80、CD83和CD86上调。在第9天成熟后观察到IL-10和IL-12水平显著升高。抗原脉冲的Mo-DC激活了抗原反应性CD8细胞毒性T细胞。在TCPS和FEP培养容器之间，未观察到Mo-DC的表面标志物表达或四聚体特异性T细胞激活效力有显著差异。
我们的研究结果表明，在小型FEP袋中产生的负载病毒抗原的Mo-DC能够引发特异性T细胞反应。鉴于临床对封闭系统DC制造的迫切需求，FEP袋是加速有前景的新兴基于DC的免疫疗法转化的一个有吸引力的选择。
