IBDV感染鸡后外周血淋巴细胞PD-1及其配体PD-L1/PD-L2转录变化的初步分析

doi:10.11843/j.issn.0366-6964.2014.04.021

Abstract

Abstract:

An assay of SYBR Green Ⅰ real-time fluorescent quantitative RT-PCR was established to investigate the expression pattern of chicken PD-l，PD-L1 and PD-L2 genes at different stages after IBDV infection.The specific primers were designed according to the sequences of PD-l，PD-L1 and PD-L2 from GenBank to amplify the objective genes for corresponding plasmids construction，and then they were used as quantitative template to construct the standard curve and melting curve for detection sensitivity，specificity and repeatability.The established SYBRS Green Ⅰ real-time fluorescent quantitative PCR (Rea1 time FQ-PCR) assay worked well with a good coefficient correlation(R²>0.99) while the template concentration was from 1×10¹-1×10⁹ copies·μL^-1，the sensitivity was sufficient to detect at least 10¹ copies of the samples.The relative transcription levels of chicken PD-l，PD-L1 and PD-L2 mRNA in PBMCs from chicken were detected on the 3rd，5th，7th and 10th day after IBDV infection by the developed Rea1 time FQ-PCR assay.At the same time，the semi quantitative RT-PCR and IBDV rapid test strip for detection were carried out to analyze the viral loads in bursal tissue.Real-time RT-PCR analysis show that the expression of PD-1 increased significantly (P<0.05) in the 7th day after infection，the expression of PD-L1 and PD-L2 increased significantly (P<0.01,P<0.05) in the 3rd day after infection and the IBDV loads in bursal tissue are highest in the 5th days after infection.The established assay was able to detect the expression of the three genes in PBMC.Our results revealing the pattern of chicken PD-l，PD-L1 and PD-L2 genes at different stages after IBDV infection，which showed that the gene transcription increased positively correlated with IBDV loads，and PD-L1 , PD-L2 increasing transcription precede PD-1.

CLC Number:

S858.315.3

WANG Ai-guo，SUN Guo-peng，LI Bo-wen，YUE Feng，ZHANG Yan-fang，ZHU Yan-ping，YIN Mei，YANG Yuan，GUO Dong-guang，WANG Xuan-nian. An Elementary Analysis for PD-l，PD-L1 and PD-L2 Genes Transcription Level of Peripheral Blood Lymphoctes in Chicken after IBDV Infection[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2014, 45(4): 654-660.

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An Elementary Analysis for PD-l，PD-L1 and PD-L2 Genes Transcription Level of Peripheral Blood Lymphoctes in Chicken after IBDV Infection

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