骨髓源肥大细胞通过甘露糖受体识别FMDV-VLPs的细胞因子应答

doi:10.11843/j.issn.0366-6964.2022.11.017

摘要/Abstract

摘要： 为了研究骨髓源肥大细胞（BMMCs）通过甘露糖受体对口蹄疫病毒样颗粒（FMDV-VLPs）的细胞因子应答效应，本研究构建了重组pCMV-HA-HBcAg-VP1-VP4质粒，并转染CHO-K1细胞以制备FMDV-VLPs。用FMDV-VLPs负载经甘露糖受体（MR）抑制剂Mannan处理的BMMCs （iMR-VLP组），并设置只负载FMDV-VLPs组（VLP组）和单纯细胞组（Control组）作为对照，收集细胞上清液，用细胞因子芯片检测不同处理组BMMCs细胞上清中细胞因子的含量。结果显示：与Control组相比，VLPs组BMMCs表达IL-1α、IL-2、IL-4、IL-15、IL-17A、IL-21、TNF-α、IFN-γ、CC17和CCL21均显著上调（P<0.05），而IL-10没有显著变化，iMR-VLP组BMMCs表达IL-1α、IL-2、IL-4、IL-15、IL-17A、TNF-α、IFN-γ、CCL-17和L-Selectin显著下调（P<0.05），而IL-9、IL-10、CCL19和CCL21的表达则呈上调变化。综上所述，FMDV-VLPs可以促进BMMCs一系列细胞因子的分泌，而抑制MR后，BMMCs分泌细胞因子能力受到了一定程度的抑制，表明FMDV-VLPs可能通过BMMCs的甘露糖受体增强Th1型细胞因子分泌、有效抑制能引起免疫抑制的IL-10的分泌，并降低介导T细胞的再循环的CCL19和CCL21的形成。研究结果为基于肥大细胞甘露糖受体应答效应的口蹄疫新型疫苗的研发提供了理论依据和新思路。

关键词: 肥大细胞, 甘露糖受体, 口蹄疫病毒样颗粒, 细胞因子, 细胞因子芯片

Abstract: To investigate the effect of bone marrow-derived mast cells (BMMCs) on the cytokines responses of FMDV-VLPs via mannose receptors, the recombinant plasmid pCMV-HA-HBcAg-VP1-VP4 was constructed and transfected into CHO-K1 cells to prepare the foot-and-mouth disease virus-like particles (FMDV-VLPs). The BMMCs treated with the mannose receptor inhibitor Mannan were loaded with FMDV-VLPs, and the BMMCs loaded with VLPs and BMMCs were used as controls. Quantitative cytokine chip was used to detect the content of cytokines secreted by BMMCs in different treatment groups. The results showed that compared with the control group, the levels of IL-1α, IL-2, IL-4, IL-15, IL-17A, IL-21, TNF-α, IFN-γ, CCL-17 and CCL21 of VLP group were all significantly up-regulated (P<0.05) and IL-10 showed no difference, while the levels of IL-1α, IL-2, IL-4, IL-15, IL-17A, TNF-α, IFN-γ, CCL-17 and L-selectin of the iMR-VLP group significantly down-regulated (P<0.05). Of note, the expression of IL-9, IL-10, CCL19 and CCL21 were further up-regulated in the iMR-VLP group. In conclusion, FMDV-VLPs can promote the secretion of a series of cytokines in BMMCs, and the ability to secrete cytokines of BMMCs is inhibited to a certain extent when MR was inhibited. The results indicate that FMDV-VLPs may enhance the secretion of Th1 cytokines through the mannose receptor of BMMCs, effectively inhibit the secretion of IL-10 that can cause immunosuppression, and reduce the formation of CCL19 and CCL21 that mediate the recycling of T cells. The results provide a theoretical basis and new ideas for the development of a new foot-and-mouth disease vaccine based on the roles of mannose receptors expressed on BMMCs.

Key words: mast cell, mannose receptor, foot-and-mouth disease virus-like particles, cytokines, cytokines chip

中图分类号:

S852.4

韩伟建, 张俊娟, 张义明, 王家鑫, 李丽敏. 骨髓源肥大细胞通过甘露糖受体识别FMDV-VLPs的细胞因子应答[J]. 畜牧兽医学报, 2022, 53(11): 3917-3926.

HAN Weijian, ZHANG Junjuan, ZHANG Yiming, WANG Jiaxin, LI Limin. Cytokine Responses of Bone Marrow-derived Mast Cells to FMDV-VLPs via Mannose Receptors[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(11): 3917-3926.

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