蜱蛋白 RHcyst-1 对鼠骨髓来源树突状细胞的免疫抑制作用。
Immunosuppressive effects of tick protein RHcyst-1 on murine bone marrow-derived dendritic cells.
机构信息
Key Laboratory of Animal Parasitology of Ministry of Agriculture, Shanghai Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Shanghai, China.
Guangdong Provincial Zoonosis Prevention and Control Key Laboratory, College of Veterinary Medicine, South China Agricultural University, Guangzhou, China.
出版信息
Parasit Vectors. 2019 Apr 15;12(1):169. doi: 10.1186/s13071-019-3411-1.
BACKGROUND
Ticks, as blood-feeding arthropod vectors, have evolved their own unique mechanism to suppress host immune responses and evade immune defenses in order to complete blood-feeding. The immunoregulatory effect of tick bioactive molecules on hosts has been widely reported, and the cystatin family has been identified as one of the major immunomodulators. In previous studies, we obtained a novel tick salivary bioactive protein named RHcyst-1, which belongs to the type 1 cystatin family. Here, we demonstrated the effects of RHcyst-1 on the host immune response mainly on dendritic cell (DC) function. Understanding the function of tick-derived bioactive molecule may help to clarify the mechanisms of how ticks escape the host immune response and help to control ticks and tick-borne disease transmission.
METHODS
Bone marrow-derived DCs (BMDCs) were generated and induced by GM-CSF and IL-4 with or without RHcyst-1 addition. Flow cytometry was used to analyze the differentiation and maturation of BMDCs and T cell cytokine production. Quantitative real-time PCR (qRT-PCR) and western blot were used to measure changes in expression within STAT and p38 MAPK signaling pathways.
RESULTS
Flow cytometry analysis revealed that RHcyst-1 inhibited the differentiation of BMDCs, but had no effect on the maturation of BMDCs. T cells co-cultured with DCs treated with RHcyst-1 produced significantly less TNF-α, IFN-γ and IL-2 than the control group. Further analysis showed that the mRNA level and phosphorylation of p38, ERK and STAT were significantly changed after RHcyst-1 added to bone marrow monocytes during the differentiation stage.
CONCLUSIONS
Our results suggest that RHcyst-1 is one of the major immunosuppressive proteins of BMDC function from blood-feeding ticks.
背景
蜱作为吸血节肢动物载体，已经进化出自己独特的机制来抑制宿主的免疫反应，逃避免疫防御，从而完成吸血。蜱生物活性分子对宿主的免疫调节作用已被广泛报道，而半胱氨酸蛋白酶抑制剂家族已被确定为主要的免疫调节剂之一。在之前的研究中，我们获得了一种新型蜱唾液生物活性蛋白，命名为 RHcyst-1，它属于 1 型半胱氨酸蛋白酶抑制剂家族。在这里，我们主要证明了 RHcyst-1 对宿主免疫反应的影响，特别是对树突状细胞（DC）功能的影响。了解蜱来源的生物活性分子的功能可能有助于阐明蜱如何逃避宿主免疫反应的机制，并有助于控制蜱和蜱传疾病的传播。
方法
用 GM-CSF 和 IL-4 诱导生成和诱导骨髓来源的树突状细胞（BMDC），并加入或不加入 RHcyst-1。流式细胞术用于分析 BMDC 的分化和成熟以及 T 细胞细胞因子的产生。实时定量 PCR（qRT-PCR）和 Western blot 用于测量 STAT 和 p38 MAPK 信号通路内表达的变化。
结果
流式细胞术分析表明，RHcyst-1 抑制 BMDC 的分化，但对 BMDC 的成熟没有影响。与对照组相比，与经 RHcyst-1 处理的 DC 共培养的 T 细胞产生的 TNF-α、IFN-γ和 IL-2 显著减少。进一步分析表明，在分化阶段向骨髓单核细胞中加入 RHcyst-1 后，p38、ERK 和 STAT 的 mRNA 水平和磷酸化明显改变。
结论
我们的结果表明，RHcyst-1 是吸血蜱对 BMDC 功能的主要免疫抑制蛋白之一。
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