Yang Yu, Wang DanYang, Li QuanXiao, He Jiang, Wang Bin, Li Jinyao, Zhang Ailian
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, Urumqi 830046, China.
Key Laboratory of Uighur Medicine, Xinjiang Institute of Materia Medica, Xinjiang 830004, China.
Vaccines (Basel). 2020 Sep 13;8(3):525. doi: 10.3390/vaccines8030525.
L. has long been used as a traditional herbal medicine owing to its immunomodulatory activity. Aqueous extracts of L. (AEAR) contain the main functional component and can activate the maturation of dendritic cells (DCs) and enhance the adaptive immunity as the adjuvant against infections. To explore the underlying mechanism of immunomodulatory activities of AEAR, DCs were produced from bone-marrow cells of mice and the effects of AEAR on cell viability were assessed by the Cell Counting Kit 8 (CCK8) method and annexin V/propidium iodide staining assays. Then, the effects of AEAR on the morphology, maturation, and function of DCs were detected using a microscope, flow cytometry-based surface receptor characterization, and endocytosis assays. The secretion levels of cytokines were then analyzed with enzyme-linked immunosorbent assay (ELISA). The activation state of DCs was evaluated by the mixed lymphocyte reaction (MLR). The activity of MAPKs and NF-κB pathways, which were involved in the regulation of AEAR on DCs, was further detected by Western blot. AEAR did not have a cytotoxic effect on DCs or mouse splenocytes. AEAR remarkably enhanced the phenotypic maturation of DCs and promoted the expression of costimulatory molecules and the secretion of cytokines in DCs. AEAR also significantly decreased the phagocytic ability of DCs and augmented the abilities of DCs to present antigens and stimulate allogeneic T-cell proliferation. Simultaneously, AEAR potently activated toll-like receptor (TLR)4-/TLR2-related MAPKs and induced the degradation of IκB and the translocation of NF-κB. In short, AEAR can profoundly enhance the immune-modulating activities of DCs via TLR4-/TLR2-mediated activation of MAPKs and NF-κB signaling pathways and is a promising candidate immunopotentiator for vaccines.
由于其免疫调节活性，L. 长期以来一直被用作传统草药。L. 的水提取物（AEAR）含有主要功能成分，可激活树突状细胞（DCs）的成熟，并作为抗感染佐剂增强适应性免疫。为了探究AEAR免疫调节活性的潜在机制，从小鼠骨髓细胞中制备DCs，并通过细胞计数试剂盒8（CCK8）法和膜联蛋白V/碘化丙啶染色试验评估AEAR对细胞活力的影响。然后，使用显微镜、基于流式细胞术的表面受体表征和内吞试验检测AEAR对DCs形态、成熟和功能的影响。随后用酶联免疫吸附测定（ELISA）分析细胞因子的分泌水平。通过混合淋巴细胞反应（MLR）评估DCs的激活状态。通过蛋白质免疫印迹进一步检测参与AEAR对DCs调节的丝裂原活化蛋白激酶（MAPKs）和核因子κB（NF-κB）信号通路的活性。AEAR对DCs或小鼠脾细胞没有细胞毒性作用。AEAR显著增强了DCs的表型成熟，促进了共刺激分子的表达和DCs中细胞因子的分泌。AEAR还显著降低了DCs的吞噬能力，并增强了DCs呈递抗原和刺激同种异体T细胞增殖的能力。同时，AEAR有力地激活了Toll样受体（TLR）4-/TLR2相关的MAPKs，并诱导IκB的降解和NF-κB的易位。简而言之，AEAR可通过TLR4-/TLR2介导的MAPKs和NF-κB信号通路的激活深刻增强DCs的免疫调节活性，是一种有前途的疫苗免疫增强剂候选物。
