ABSTRACT

1. Chicken erythrocytes in blood vessels are the most abundant circulating cells, which participate in the host’s immune responses. The transcription factor nuclear factor-kappa B (NF-κB) plays a vital role in the inflammatory response following viral infections. However, the expression of the NF-κB pathway, and other immune-related genes in chicken erythrocytes infected with low pathogenic avian influenza virus (LPAIV H9N2), has not been extensively studied.

2. The following study determined the interaction of LPAIV H9N2 with chicken erythrocytes using indirect immunofluorescence microscopy. This was followed by investigating myeloid differentiation primary response 88 (MyD88), C-C motif chemokine ligand 5 (CCL5), melanoma differentiation-associated protein 5 (MDA5), the inhibitor of nuclear factor-kappa B kinase subunit epsilon (IKBKE), NF-κB inhibitor alpha (NFKBIA), NF-κB inhibitor epsilon (NFKBIE), interferon-alpha (IFN-α), colony-stimulating factor 3 (CSF3) and tumour necrosis factor receptor-associated factor 6 (TRAF6) by mRNA expression using quantitative real-time PCR (qRT-PCR) at four different time intervals (0, 2, 6 and 10 h).

3. There was a significant interaction between erythrocytes and LPAIV H9N2 virus. Furthermore, the mRNA expression of the NF-κB pathway and other immune-related genes were significantly up-regulated at 2 h post-infection in infected chicken erythrocytes, except for TRAF6, which were significantly downregulated. While at 0 h post-infection, IFN-α and CSF3 were significantly upregulated, whereas NFKBIA was significantly downregulated. Further expression of MDA5, CCL5 and NFKBIA was upregulated, while TRAF6 was downregulated at 6 h post-infection. In infected erythrocytes, expression of MyD88, CCL5 and IKBKE was upregulated. However, IFN-α and TRAF6 were downregulated at 10 h post-infection.

4. These results give initial evidence that the NF-κB pathway, and other genes related to immunity, in chicken erythrocytes may contribute to LPAIV subtype H9N2 and induce host immune responses.

摘要

1. 血管中的鸡红细胞是最丰富的循环细胞，参与宿主的免疫反应。转录因子核因子-κB (NF-κB) 在病毒感染后的炎症反应中起着至关重要的作用。然而，NF-κB 通路和其他免疫相关基因在感染低致病性禽流感病毒 (LPAIV H9N2) 的鸡红细胞中的表达尚未得到广泛研究。

2. 以下研究使用间接免疫荧光显微镜确定 LPAIV H9N2 与鸡红细胞的相互作用。随后研究了髓系分化初级反应 88 ( MyD88 )、CC 基序趋化因子配体 5 ( CCL5 )、黑色素瘤分化相关蛋白 5 ( MDA5 )、核因子-κB 激酶亚基 epsilon ( IKBKE )抑制剂、NF- κB抑制剂α（NFKBIA）、NF-κB抑制剂ε（NFKBIE）、干扰素α（IFN-α）、集落刺激因子3（CSF3）和肿瘤坏死因子受体相关因子6（TRAF6）通过mRNA在四个不同的时间间隔（0、2、6 和 10 小时）使用定量实时 PCR (qRT-PCR) 进行表达。

3.红细胞与LPAIV H9N2病毒之间存在显着的相互作用。此外，NF-κB 通路和其他免疫相关基因的mRNA表达在感染后 2 小时内在感染的鸡红细胞中显着上调，但TRAF6除外，其显着下调。而在感染后 0 小时，IFN-α和CSF3显着上调，而NFKBIA显着下调。MDA5、CCL5和NFKBIA的进一步表达上调，而TRAF6在感染后6小时下调。在受感染的红细胞中，MyD88 的表达、CCL5和IKBKE被上调。然而，IFN-α和TRAF6在感染后 10 小时被下调。

4. 这些结果初步证明了鸡红细胞中的 NF-κB 通路和其他与免疫相关的基因可能有助于 LPAIV 亚型 H9N2 并诱导宿主免疫反应。