口蹄疫病毒样颗粒诱导的肥大细胞活化对IgG应答的影响  

Effect of mast cells activation induced by foot-and-mouth disease virus-like particles on IgG immune responses in mice

作　　者：刘佳欢 崔川 张俊娟 韩伟建 张义明[1] 李丽敏[1] 王家鑫[1] LIU Jiahuan;CUI Chuan;ZHANG Junjuan;HAN Weijian;ZHANG Yiming;LI Limin;WANG Jiax-in(College of Veterinary Medicine,Hebei Agricultural University,Baoding,Hebei 071000,China)

机构地区：[1]河北农业大学动物医学院,河北保定071000

出　　处：《中国兽医学报》2021年第10期1983-1989,共7页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31402174);河北省教育厅重点基金资助项目(ZD2015040);河北省青年拔尖人才基金资助项目。

摘　　要：为研究口蹄疫病毒样颗粒(foot-and-mouth disease virus-like particles,FMDV VLP)诱导的肥大细胞(mast cells,MCs)脱颗粒对IgG应答的影响,用FMDV VLP免疫小鼠(VLP组),同时设置PBS对照组,首次免疫后14 d进行加强免疫,分别在14 dpi(days post immunization)和28 dpi,取2组小鼠皮肤、颌下淋巴结、脾脏与颌下腺,用恒温冷冻切片机制备冰冻切片,经甲苯胺蓝染色观察MCs脱颗粒现象;采用ELISA检测小鼠血清中TNF-α和IL-10以及FMDV特异性IgG抗体的水平。结果显示,FMDV-VLP可在体内诱导小鼠MCs脱颗粒,14 dpi时VLP组主要是皮肤表皮内的MCs脱颗粒和淋巴结中被膜下窦的MCs脱颗粒;28 dpi时,VLP组主要是皮下组织中的MCs和淋巴结深层髓质的MCs脱颗粒。14 dpi VLP组小鼠皮肤(P<0.01)、淋巴结(P<0.01)和脾脏(P<0.05)中MCs活化数量分别显著高于14 dpi PBS组;而28 dpi VLP组小鼠皮肤(P<0.01)、淋巴结(P<0.01)、脾脏(P<0.01)与颌下腺(P<0.05)中MCs活化数量均显著高于28 dpi PBS组。VLP组小鼠血清中FMDV特异性IgG水平在14 dpi呈阴性,而在28 dpiIgG水平呈阳性。此外,在14 dpi和28 dpi,VLP组小鼠血清中TNF-α水平均显著高于PBS组(P<0.05),而血清中IL-10水平与PBS组相比均没有统计学差异。以上结果表明,FMDV VLP可以诱导小鼠皮肤、淋巴结、脾脏和颌下腺中的MCs活化,同时可以促进机体分泌TNF-α,提示MCs颗粒成分可能在VLP诱导小鼠产生IgG免疫应答过程中发挥了某种促进作用。本研究为FMDV疫苗新型佐剂的研发及免疫应答机制研究提供了理论基础。To study the effect of foot-and-mouth disease virus-like particles(FMDV VLP)on mast cells(MCs)degranulation and the immune response in mice,mice were immunized with FMDV VLP(VLP group),meanwhile,PBS control group was set,booster immunization was performed 14 d after initial immunization.Skin,submandibular lymph nodes,spleen,and mandibular glands of the two groups were taken for frozen sections at 14 dpi and 28 dpi,respectively.MCs degranulation was observed by toluidine blue staining.ELISA was used to detect the levels of FMDV-specific IgG,TNF-αand IL-10 in serum.The results showed that FMDV-VLP could induce MCs degranulation in vivo,and at 14 dpi,the degranulation of MCs in skin epidermis and in subcapsular sinus of lymph nodes were mainly observed in VLP group;at 28 dpi,the degranulation of MCs in subcutaneous tissue and in deep medulla of lymph nodes were mainly observed in VLP group.The number of MCs activation in the skin(P<0.01),lymph node(P<0.01)and spleen(P<0.05)at 14 dpi in VLP group was significantly higher than that in PBS group,respectively;while the number of MCs activation in the skin(P<0.01),lymph node(P<0.01),spleen(P<0.01)and submandibular gland(P<0.05)at 28 dpi in VLP group was significantly higher than that in PBS group,respectively.In addition,at both 14 dpi and 28 dpi,serum levels of TNF-αin VLP group was significantly higher than that in PBS group after booster immunization(P<0.05),however,there was no statistical difference in IL-10 level between the VLP group and the PBS group.Serum levels of FMDV-specific IgG were negative at 14 dpi and positive at 28 dpi.The above results indicated that FMDV VLP could successfully induce degranulation of MCs in skin,lymph nodes,spleen and submandibular gland of mice,and immunization with VLP could promote TNF-αsecretion,but had no effect on the secretion of IL-10,suggesting that the degranulation products of MCs may play an important role in promoting IgG immune response induced by VLP,which provides a theoretical basis for studying the novel adjuvant

关 键 词：肥大细胞 口蹄疫病毒样颗粒 细胞因子 IGG 

分 类 号：S853.1[农业科学—临床兽医学] R535[农业科学—兽医学]