Arromsava Artittaya, Chuchawankul Siriporn, Worasilchai Navaporn, Angkasekwinai Pornpimon, Amornsupak Kamolporn
Program of Molecular Sciences in Medical Microbiology and Immunology, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
Department of Transfusion Medicine and Clinical Microbiology, Faculty of Allied Health Sciences, Chulalongkorn University, Bangkok, 10330, Thailand.
BMC Complement Med Ther. 2025 Jul 9;25(1):251. doi: 10.1186/s12906-025-04990-z.
Treatment of Cryptococcus neoformans infection is still challenging due to high side effects and acquired drug resistance. Eliminating fungal infection requires monocytes and macrophages, which are weakened by the non-protective type-2 immune response induced by the pathogen. Pleurotus pulmonarius (PP) is an edible mushroom rich in bioactive molecules, particularly β-glucan. This study aimed to investigate the immunomodulatory activities of PP crude extract on monocytes and macrophages against C. neoformans.
PP was extracted using a hot aqueous method and precipitated with absolute ethanol. Glucan content was assessed by colorimetric assay. The cytotoxicity of PP crude extract was tested using an MTT assay. The immunomodulatory activities and preventive potential of PP crude extract for C. neoformans infection were investigated using human THP-1 monocyte and THP-1-derived macrophage models. Proinflammatory and anti-inflammatory cytokine production was evaluated using RT-qPCR and ELISA. Immune mechanisms related to C. neoformans clearance were studied, including phagocytosis, ROS production, macrophage polarization, and antifungal killing.
The result showed that PP crude extract contains α-glucan (14.72% w/w) and β-glucan (11.75% w/w). The immunomodulatory activities of PP crude extract on the expressions of TNFA, IL1B, IL6, and MCP1/CCL2 were observed in THP-1 cells. This finding corresponds to the significant increase in TNF-α level measured by ELISA. Moreover, PP crude extract enhanced the release of TNF-α and IL-10 from THP-1-derived macrophages. Interestingly, pre-exposure to PP crude extract elicits preventive potential for controlling C. neoformans infection. A significant increase in type 1 cytokines was observed in C. neoformans-infected monocytes and macrophages. While C. neoformans induced M2 polarization, this phenotype was attenuated in PP-trained macrophages partly through MCP1/CCL2 expression and CD206 M2 suppression. Moreover, 100 µg/mL of PP crude extract pretreatment could impair C. neoformans intracellular proliferation in THP-1-derived macrophages and upregulate ROS production.
This study highlights the immunomodulatory potential of PP crude extract as a plant-based immunomodulator to enhance protective antimicrobial responses, particularly through ROS-mediated mechanisms of antifungal immunity. These findings support its potential application in developing natural therapeutic supplements, functional foods, and trained immunity-based strategies as alternative approaches to strengthening host defense against infections.
新型隐球菌感染的治疗仍然具有挑战性，因为其副作用大且存在获得性耐药。消除真菌感染需要单核细胞和巨噬细胞，而病原体诱导的非保护性2型免疫反应会削弱这些细胞的功能。肺侧耳（PP）是一种富含生物活性分子，特别是β-葡聚糖的食用菌。本研究旨在探讨PP粗提物对单核细胞和巨噬细胞抗新型隐球菌的免疫调节活性。
采用热水提取法提取PP，并用无水乙醇沉淀。通过比色法测定葡聚糖含量。使用MTT法检测PP粗提物的细胞毒性。利用人THP-1单核细胞和THP-1衍生的巨噬细胞模型，研究PP粗提物对新型隐球菌感染的免疫调节活性和预防潜力。使用RT-qPCR和ELISA评估促炎和抗炎细胞因子的产生。研究了与新型隐球菌清除相关的免疫机制，包括吞噬作用、活性氧产生、巨噬细胞极化和抗真菌杀伤。
结果表明，PP粗提物含有α-葡聚糖（14.72% w/w）和β-葡聚糖（11.75% w/w）。在THP-1细胞中观察到PP粗提物对TNFA、IL1B、IL6和MCP1/CCL2表达的免疫调节活性。这一发现与ELISA检测到的TNF-α水平显著升高一致。此外，PP粗提物增强了THP-1衍生巨噬细胞中TNF-α和IL-10的释放。有趣的是，预先暴露于PP粗提物可引发控制新型隐球菌感染的预防潜力。在新型隐球菌感染的单核细胞和巨噬细胞中观察到1型细胞因子显著增加。虽然新型隐球菌诱导了M2极化，但在PP预处理的巨噬细胞中，这种表型部分通过MCP1/CCL2表达和CD206 M2抑制而减弱。此外，100 μg/mL的PP粗提物预处理可损害新型隐球菌在THP-1衍生巨噬细胞中的细胞内增殖，并上调活性氧的产生。
本研究强调了PP粗提物作为一种基于植物的免疫调节剂的免疫调节潜力，可增强保护性抗菌反应，特别是通过活性氧介导的抗真菌免疫机制。这些发现支持了其在开发天然治疗补充剂、功能性食品以及基于训练免疫的策略方面的潜在应用，作为加强宿主抗感染防御的替代方法。
