Laboratory of Medical Investigation in Dermatology and Immunodeficiencies (LIM-56) Clinical Hospital HCFMUSP Faculty of Medicine University of Sao Paulo, Sao Paulo, SP, Brazil.
Electron Microscopy Nucleus Adolfo Lutz Institute, Sao Paulo, Brazil.
J Immunol Res. 2024 Jun 7;2024:6817965. doi: 10.1155/2024/6817965. eCollection 2024.
Therapeutic vaccines based on monocyte-derived dendritic cells have been shown to be promising strategies and may act as complementary treatments for viral infections, cancers, and, more recently, autoimmune diseases. Alpha-type-1-polarized dendritic cells (aDC1s) have been shown to induce type-1 immunity with a high capacity to produce interleukin-12p70 (IL-12p70). In the clinical use of cell-based therapeutics, injectable solutions can affect the morphology, immunophenotypic profile, and viability of cells before delivery and their survival after injection. In this sense, preparing a cell suspension that maintains the quality of aDC1s is essential to ensure effective immunotherapy. In the present study, monocytes were differentiated into aDC1s in the presence of IL-4 and GM-CSF. On day 5, the cells were matured by the addition of a cytokine cocktail consisting of IFN-, IFN-, IL-1, TNF-, and Poly I:C. After 48 hr, mature aDC1s were harvested and suspended in two different solutions: normal saline and Ringer's lactate. The maintenance of cells in suspension was evaluated after 4, 6, and 8 hr of storage. Cell viability, immunophenotyping, and apoptosis analyses were performed by flow cytometry. Cellular morphology was observed by electron microscopy, and the production of IL-12p70 by aDC1s was evaluated by ELISA. Compared with normal saline, Ringer's lactate solution was more effective at maintaining DC viability for up to 8 hr of incubation at 4 or 22°C.
基于单核细胞衍生的树突状细胞的治疗性疫苗已被证明是一种很有前途的策略,可作为病毒感染、癌症的辅助治疗方法,最近还可作为自身免疫性疾病的辅助治疗方法。α-1 型极化树突状细胞(aDC1)被证明具有诱导 1 型免疫的能力,能够高表达白细胞介素-12p70(IL-12p70)。在细胞治疗的临床应用中,可注射溶液会影响细胞在输送前的形态、免疫表型特征和活力,以及注射后的存活能力。从这个意义上说,制备能维持 aDC1 质量的细胞悬浮液对于确保有效的免疫治疗至关重要。在本研究中,单核细胞在 IL-4 和 GM-CSF 的存在下分化为 aDC1。第 5 天,通过添加由 IFN-、IFN-、IL-1、TNF-和 Poly I:C 组成的细胞因子鸡尾酒使细胞成熟。48 小时后,收获成熟的 aDC1 并悬浮在两种不同的溶液中:生理盐水和林格氏乳酸盐。在储存 4、6 和 8 小时后评估细胞在悬浮液中的维持情况。通过流式细胞术进行细胞活力、免疫表型和细胞凋亡分析。通过电子显微镜观察细胞形态,通过 ELISA 评估 aDC1 产生的 IL-12p70。与生理盐水相比,林格氏乳酸盐溶液在 4°C 或 22°C 孵育长达 8 小时时更有效地维持 DC 活力。