摘要
本研究旨在阐明鸡氨肽酶N(chAPN)、唾液酸以及硫酸乙酰肝素在传染性支气管炎病毒(IBV)M41株感染宿主细胞中的作用以及3种受体特异抑制剂对IBV M41株在自然宿主CEK细胞内增殖能力的影响。作者选择苯丁抑制素(Bestatin)、神经氨酸酶(NA)和肝素酶Ⅲ分别作为APN、唾液酸以及硫酸乙酰肝素的抑制剂,在不同条件下,单独或共同预处理CEK细胞,而后接入IBV M41株病毒感染细胞,应用荧光定量PCR方法定量检测IBVM41株在CEK细胞中的增殖变化,应用鸡胚半数感染剂量法(EID50)测定病毒感染鸡胚能力的变化。结果表明,经Bestatin和NA处理的CEK细胞获得了抵抗IBV M41株感染的能力,与未经处理的(对照组)细胞相比,Besta-tin和NA能显著降低IBV M41株在CEK细胞内的增殖及对鸡胚的感染能力(P<0.01),且Bestatin处理后CEK细胞内的病毒增殖量显著低于NA处理后CEK细胞内的病毒增殖量,两者病毒液的EID50滴定值差异显著(P<0.01);肝素酶Ⅲ的处理则对病毒增殖无显著影响;3种受体抑制剂共同处理CEK细胞后,病毒增殖量显著下降(P<0.01),但仍有病毒增殖,病毒液的EID50滴定值为102.12±0.05.0.1mL-1。结果提示,chAPN在IBV感染宿主细胞的过程中发挥特异性受体作用,而唾液酸在IBV感染宿主细胞中发挥辅助受体作用,硫酸乙酰肝素不是IBV在自然感染中的必需因素,同时提示,可能存在其他未知受体因子参与IBV感染宿主细胞的过程。
This experiment was conducted to research the roles of aminopeptidase N,sialic acid and heparin sulfate during infectious bronchitis virus（IBV） entry into host cells,and the inhibition of the possible receptor inhibitors during the progress of the virus replication in the chicken embryo kidney（CEK） cells.Bestatin,NA and heparinase Ⅲ are chosen as the inhibitors of aminopeptidase N,sialic acid and heparin sulfate to pre-treat CEK cells alone or collaboratively in different conditions,and then IBV was infected into prepared CEK cells.The proliferation of IBV was detected by FQ-PCR.Virus titers were determined as 50% egg infectious doses（EID50） per gram of tissue using the Reed-Muench method.The result showed that,compared with the control group,the CEK cells treated by Bestatin and Neuraminidase acquired resistance to IBV M41 strain,which can significantly reduce the proliferation of IBV M41 strain（P0.01）,the inhibitory effects of Bestatin were stronger than that of Neuraminidase（P0.01）.Heparinase Ⅲ treatment couldn＇t influence the susceptibility of IBV M41 strain in CEK cells.50% egg infective dose of infectious bronchitis virus strain M41 from different kinds of sources was different.The ability of the viral multiplication significantly decreased（P0.01） in the CEK cells treated with these receptor inhibitors,but didn＇t prohibit the viral multiplication,the EID50 value was 102.12±0.05·0.1 mL-1.From the experimental results,we can speculate that chicken aminopeptidase N is more critical than sialic acid during the natural infection of IBV M41 strain,but they both act as the functional cellular receptor during the progress of the virus infection,while the heparin sulfate isn＇t the functional cellular receptor.There are also another unknown receptors in the progress of infection.So it is important to find other unknown receptors and use the receptor inhibitors to prevent and cure infectious bronchitis virus.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2011年第5期704-710,共7页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
黑龙江省教育厅科学技术研究项目(2003fz012)
东北农业大学创新团队课题(CXT006-4-1)
