An Investigation of the Innate Immune Response in Bovine Mammary Epithelial Cells Challenged by Prototheca zopfii
An Investigation of the Innate Immune Response in Bovine Mammary Epithelial Cells Challenged by Prototheca zopfii
复制标题
牛乳腺上皮细胞对 Prototheca zopfii 攻击的先天免疫反应的研究
DOI：
10.1007/s11046-016-0053-0
复制
发表时间：
2016-12-01
期刊：
影响因子：
5.5
通讯作者：
Han, Bo
中科院分区：
文献类型：
--
作者：
Deng, Zhaoju;Shahid, Muhammad;Han, Bo
zoopfiprotocaa是一种重要的牛乳腺炎病原体，可导致严重的乳腺感染。然而，牛乳腺炎与猪瘟假单胞菌相关的先天免疫反应尚不清楚。因此，本研究的目的是探讨猪瘟假单胞菌对体外先天免疫反应的影响。分别用5.0 × 10(4)个细胞/ml的基因型感染牛乳腺上皮细胞（bMECs），采用实时聚合酶链反应（RT-PCR）检测1、3、6、12和24 h内TLR-2、TLR-4、tnf - α、IL-1 β、IL-8、NOD-1、NOD-2和β -防御素-5的mRNA表达，并检测同一时间内感染牛乳腺上皮细胞细胞核和细胞质中nf - κ B p65蛋白的含量。结果表明，与基因型相比，zopfii基因型对TLR-2、TLR-4、tnf - α、IL-1 β、IL-8、NOD-1、NOD-2和β -防御素-5′的表达上调能力更强。Western blot结果显示，zopfii基因型攻毒后，bMECs细胞核内NF-kappa B p65蛋白表达上调，细胞质内NF-kappa B p65蛋白表达抑制；这表明bmec在被II型zopfii感染时，部分通过nf - κ B信号通路调节先天免疫反应和炎症。由此可见，基因II型的粘附力强于基因I型，因此基因II型的调控能力比基因I型更强，从而引起牛乳腺组织的炎症反应。
Prototheca zopfii is an important bovine mastitis pathogen, which could result in severe mammary infection. However, the innate immune response in bovine mastitis associated with P. zopfii was not clear. Therefore, the aim of this study is to investigate in vitro innate immune responses implicated by P. zopfii. Bovine mammary epithelial cells (bMECs) were infected with 5.0 x 10(4) cells/ml P. zopfii genotypes I and II independently, and the mRNA expression of TLR-2, TLR-4, TNF-alpha, IL-1 beta, IL-8, NOD-1, NOD-2 and beta-defensin-5 was determined by real-time polymerase chain reaction (RT-PCR) over a time course of 1, 3, 6, 12 and 24 h. The detection of the NF-kappa B p65 protein in nucleus and cytoplasm of infected bMECs over the same time course was evaluated. Results showed that P. zopfii genotype II has ability to up-regulate the expression of TLR-2, TLR-4, TNF-alpha, IL-1 beta, IL-8, NOD-1, NOD-2 and beta-defensin-5 'more strongly than genotype I. Western blot results showed that when bMECs were challenged by P. zopfii genotype II, the expression of NF-kappa B p65 protein in the nucleus was up-regulated, while in cytoplasm it appeared to be repressed, which indicated that bMECs partly regulate the innate immune responses and inflammation by the NF-kappa B signaling pathway while being infected by P. zopfii genotype II. It was concluded that adhesion of genotype II was stronger than genotype I, and therefore the genotype II regulatory ability is more robust than that of the genotype I, which causes inflammation of bovine mammary tissue.