Induction of avian β-defensins by CpG oligodeoxynucleotides and proinflammatory cytokines in hen vaginal cells in vitro
Induction of avian β-defensins by CpG oligodeoxynucleotides and proinflammatory cytokines in hen vaginal cells in vitro
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DOI：
10.1530/rep-12-0518
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发表时间：
2013-06-01
期刊：
影响因子：
3.8
通讯作者：
Yoshimura, Yukinori
中科院分区：
文献类型：
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作者：
Sonoda, Yuka;Mageed, Ahmad M. Abdel;Yoshimura, Yukinori
鸡输卵管阴道的免疫功能是防止泄殖腔微生物感染的必要条件。本研究的目的是确定CpG寡脱氧核苷酸（CpG-ODN）是否刺激鸡阴道细胞中禽β-防御素（AvBDs）的表达。具体问题是CpG-ODN是否影响AvBD和促炎细胞因子的表达，以及细胞因子是否影响阴道细胞中AvBD的表达。对分散的白色来航产蛋母鸡阴道细胞进行培养，并用不同剂量的脂多糖（LPS）、CpG-ODN、白细胞介素1 β（IL 1B）或IL 6进行刺激。培养的细胞群含有上皮细胞、成纤维细胞样细胞和CD 45阳性白细胞。AvBD 3、AvBD-10和AvBD-12的免疫反应性定位于阴道切片的粘膜上皮中。用定量RT-PCR分析AvBD、IL 1B和IL 6的表达。RT-PCR分析显示，在无刺激的培养阴道细胞中，AvBD 1、-3、-4、-5、-10和-12均有表达。Toll样受体4和Toll样受体21分别识别LPS和CpG-ODN，IL 1和IL 6受体（IL 1 R 1和IL 6 R）也在它们中表达。LPS可上调IL 1B、IL 6、AvBD 10和AvBD-12的表达，而CpG-ODN仅上调IL 1B和IL 6的表达。IL 1B刺激上调AvBD 1和-3表达，而IL 6刺激并未引起AvBD表达的变化。这些结果表明，微生物来源的CpG-ODN通过与TLR 21相互作用上调IL 1B和IL 6的表达，然后IL 1B诱导AvBD 1和AvBD-3来预防阴道感染。
Immune function in the vagina of hen oviduct is essential to prevent infection by microorganisms colonizing in the cloaca. The aim of this study was to determine whether CpG oligodeoxynucleotides (CpG-ODN) stimulate the expression of avian beta-defensins (AvBDs) in hen vaginal cells. Specific questions were whether CpG-ODN affects the expression of AvBDs and proinflammatory cytokines and whether the cytokines affect AvBDs expression in vaginal cells. The dispersed vaginal cells of White Leghorn laying hens were cultured and stimulated by different doses of lipopolysaccharide (LPS), CpG-ODN, interleukin 1 beta (IL1B), or IL6. The cultured cell population contained epithelial cells, fibroblast-like cells, and CD45-positive leukocytes. The immunoreactive AvBD3, -10, and -12 were localized in the mucosal epithelium in the section of the vagina. The expression of AvBDs, IL1B, and IL6 was analyzed by quantitative RT-PCR. RT-PCR analysis showed the expression of AvBD1, -3, -4, -5, -10, and -12 in the cultured vaginal cells without stimulation. Toll-like receptors (TLRs) 4 and 21, which recognize LPS and CpG-ODN respectively and IL1 and IL6 receptors (IL1R1 and IL6R) were also expressed in them. The expression of IL1B, IL6, and AvBD10 and -12 was upregulated by LPS, whereas only IL1B and IL6 were upregulated by CpG-ODN. IL1B stimulation upregulated AvBD1 and -3 expression, whereas IL6 stimulation did not cause changes in AvBDs expression. These results suggest that CpG-ODN derived from microbes upregulates the expression of IL1B and IL6 by interaction with TLR21 and then IL1B induces AvBD1 and -3 to prevent infection in the vagina.