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| Effects of Chicken (Gallus gallus) B-Fα Gene Silencing on Transcription Level of IL-2, IL-4 and IL-6 |
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Abstract Major histocompatibility complex (MHC) and interleukin (IL) are important immune molecules, in which MHC classⅠ molecules present endogenous antigen and induce initial immune response, and ILs are critical immune regulators. Little information about relationship between chicken (Gallus gallus) MHC classⅠ(also named as B-Fα) and IL genes is reported. The present study focused on effects of downgraded expression of B-Fα on IL genes using the method of gene silencing. First, according to the sequence of B-Fα gene and the structure require of shRNA (short hairpin RNA) gene sequence, 3 shRNA fragments with potential gene silencing were selected and synthesized, and inserted into lentiviruse expression plasmids pLL3.7 with restriction enzyme, respectively. The 3 recombinant plasmids were then transfected into 293T cells, which could express label protein in the cells and also be packed into lentiviruses. The recombinant viruses could infect HD-11 cells with efficiency of over 95%, and the gene transcription levels in the HD-11 cells detected by qRT-PCR were downgraded to 39%, 97% and 61% respectively. Finally the effects of B-Fα gene silencing on the transcription levels of cytokine genes IL-2, IL-4, and IL-6 were detected with recombinant lentiviruses (containing pLL-shB-Fα-257), which had the strongest silencing (downgraded to 39%). The results showed that in the cells with down regulated B-Fα gene, the transcription levels of IL were influenced: IL-2 was down-regulated (P＜0.01), IL-4 was up-regulated (P＜0.05), and IL-6 had no change (P＞0.05). All above results indicated that B-Fα gene transcription could be interfered by the gene silencing method, while down regulation of B-Fα gene expression influenced IL transcription level. These suggest that there might be critical association between B-Fα genes, which inducing cellular immune responses, and cytokine genes which are the downstream regulatory factors. The present results provide basic data for further study about regulation mechanism of immune responses.
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Received: 25 January 2018
Published: 26 September 2018
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