Effects of host fatty acid-binding protein 4 on Eimeria tenella sporozoites invasion of cells
Effects of host fatty acid-binding protein 4 on Eimeria tenella sporozoites invasion of cells
复制标题
宿主脂肪酸结合蛋白4对柔嫩艾美耳球虫子孢子侵袭细胞的影响
DOI：
10.1007/s00436-019-06321-x
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发表时间：
2019-06-01
影响因子：
2
通讯作者：
Dong, Hui
中科院分区：
文献类型：
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作者：
Wang, Lu;Zhu, Shunhai;Dong, Hui
在我们之前的研究中，通过对感染柔嫩艾美耳球虫子孢子的宿主细胞进行蛋白质组学分析，结合等压标记进行相对和绝对定量，鉴定了几种与艾美耳球虫入侵相关的宿主蛋白。本研究将鸡脂肪酸结合蛋白4(FABP4)基因克隆到原核表达载体pET-28c(+)中，构建了原核表达载体pET-28c(+)-FABP4。表达了18.5 kDa的重组FABP4蛋白(RFABP4)，并进行了Western blotting鉴定。免疫印迹和免疫组织化学检测柔嫩艾美耳球虫子孢子感染的DF-1细胞中FABP4的表达明显低于未感染的细胞。抗体抑制实验表明，50、100、200、300、400g/mLFABP4抗体对子孢子侵袭无明显影响。分别用BMS-309403和转化生长因子-3抑制和促进FABP4在DF-1细胞中的表达，并用流式细胞仪检测它们对子孢子侵袭细胞的影响。BMS-309403处理组子孢子侵袭率无明显影响，而转化生长因子-3处理组子孢子侵袭率显著下降。这些结果表明，宿主FABP4在艾美耳球虫入侵过程中起着负性作用。然而，FABP4负性调控艾美耳球虫侵袭的确切机制还需要进一步的研究。
In our previous study, proteomics analyses of host cells infected with Eimeria tenella sporozoites coupled with isobaric tags for relative and absolute quantitation, identified several host proteins related to Eimeria invasion. In this study, A 458-bp Gallus gallus fatty acid-binding protein 4 (FABP4) gene was cloned and subcloned to pET-28c(+) vector to construct the prokaryotic recombinant expression plasmid pET-28c(+)-FABP4. The 18.5kDa recombinant FABP4 protein (rFABP4) was expressed and identified by western blotting. Expression of FABP4 in E. tenella sporozoite-infected DF-1 cells was downregulated significantly than in non-infected cells detected by western blotting and immunohistochemistry. The antibody inhibition assay showed that antibodies against FABP4 at 50, 100, 200, 300, and 400g/mL had no significant effect on sporozoite invasion. BMS-309403 and transforming growth factor-3 (TGF-3) was used to inhibit and improve the expression of FABP4 in DF-1 cells, respectively, and their effect on the sporozoite invasion of cells was detected by flow cytometry. Sporozoite invasion rate in the BMS-309403-treated group was not significantly affected; however, the invasion rate in the TGF-3-treated group declined significantly. These results show that host FABP4 plays a negative role in Eimeria invasion. However, further studies are needed to elucidate the exact mechanism of how FABP4 negatively regulates Eimeria invasion.