猪肺泡巨噬细胞极化对猪瘟病毒复制的影响

摘要

为探究猪肺泡巨噬细胞(PAM)极化与猪瘟病毒(CSFV)复制之间的关系,本研究首先通过支气管肺泡灌洗液法建立了高纯度原代肺泡巨噬细胞的培养体系.通过western blot确立了PAM向M1和M2型巨噬细胞极化的鉴别体系,并用CCK8法检测巨噬细胞极化后的细胞活性.在此基础上,通过间接免疫荧光试验和荧光定量PCR分别检测感染不同亚型PAM的CSFV E2蛋白的表达及其E2基因的转录水平,显示PAM向M1型的极化可减少CSFV E2蛋白的表达,从而抑制病毒的复制.本研究不仅为研究宿主对CSFV的免疫机制提供了重要线索,而且为设计基于巨噬细胞极化亚型防制CSF新策略提供一定依据.%To explore the relationship between porcine alveolar macrophage (PAM) polarization and classical swine fever virus (CSFV) replication.Firstly,the culture system for highly pure primary alveolar pulmonary macrophages was established by the method of porcine bronchoalveolar lavage fluid.Secondly,the identification system for alveolar macrophage to M1 and M2 polarization by western blot,and the viability of different subtype PAM was detected by CCK-8.Based on the above methods,the expression of CSFV E2 protein and the transcriptional level of CSFV in CSFV infected different sultypes PAM CSFV in different subtypes of PAM infected with CSFV were detected by indirect immunofluorescence assay and real-time PCR.The results showed that the polarization of macrophages to M 1 could inhilit the expression of CSFV E2 protein,thereby inhibiting the viral replication.This study not only provides an important clue for studying the immune mechenism of host to CSFV,but also provides a basis for the design of the new strategy for controlling CSFV based on macrophage polarization subtype.