CAJ | 학술논문

聚肌胞(Poly I：C)是一种天然双链RNA(Double strand RNA, dsRNA)的拟似物，能够模拟病毒感染后所形成的 dsRNA 及刺激机体产生抗病毒免疫反应。文章以抗病力存在差异的大蒲莲和长白仔猪为研究对象，分离外周血单核细胞(Peripheral blood mononuclear cell, PBMC)，在20μg/mL的Poly I：C的免疫刺激下体外培养24 h，对影响免疫应答过程中的7个细胞因子(IRF3、IL6、IL8、IL10、TNFα、IFNγ和IFNα)和3个模式识别受体(TLR3、TLR4和RIG1)利用实时荧光定量PCR检测Poly I：C免疫刺激组相对于对照组的基因表达变化倍数。结果表明：检测的大部分细胞因子和受体(6个)表达量变化倍数很大，其中3种白细胞介素IL6、IL8和IL10免疫刺激变化倍数最大，平均变化倍数分别为20.71、10.87和5.18倍。对不同个体和品种间的比较发现，不仅大蒲莲和长白两品种间(大蒲莲猪的变化倍数平均高于长白猪)而且同品种的3头全同胞仔猪间对Poly I：C免疫刺激的应答也存在较大的变化。文章利用Poly I：C体外模拟dsRNA对PBMC的感染，为下一步筛选仔猪对Poly I：C刺激的免疫应答基因及鉴定大蒲莲猪特殊的抗性基因奠定了基础。
취기포(Poly I：C)시일충천연쌍련RNA(Double strand RNA, dsRNA)적의사물，능구모의병독감염후소형성적 dsRNA 급자격궤체산생항병독면역반응。문장이항병력존재차이적대포련화장백자저위연구대상，분리외주혈단핵세포(Peripheral blood mononuclear cell, PBMC)，재20μg/mL적Poly I：C적면역자격하체외배양24 h，대영향면역응답과정중적7개세포인자(IRF3、IL6、IL8、IL10、TNFα、IFNγ화IFNα)화3개모식식별수체(TLR3、TLR4화RIG1)이용실시형광정량PCR검측Poly I：C면역자격조상대우대조조적기인표체변화배수。결과표명：검측적대부분세포인자화수체(6개)표체량변화배수흔대，기중3충백세포개소IL6、IL8화IL10면역자격변화배수최대，평균변화배수분별위20.71、10.87화5.18배。대불동개체화품충간적비교발현，불부대포련화장백량품충간(대포련저적변화배수평균고우장백저)이차동품충적3두전동포자저간대Poly I：C면역자격적응답야존재교대적변화。문장이용Poly I：C체외모의dsRNA대PBMC적감염，위하일보사선자저대Poly I：C자격적면역응답기인급감정대포련저특수적항성기인전정료기출。
Polyinosinic-polycytidylic acid (Poly I:C) is an analogue of natural double strand RNA (dsRNA), which can simulate the viral dsRNA and stimulate the immune response. In the present study, peripheral blood mononuclear cells (PBMC) were isolated from piglets of Dapulian and Landrace with different disease resistance, and stimulated with 20μg/mL Poly I:C for 24 hours in vitro culture. The expression of several cytokines (IL6, IL8, TNFα, IL10, IRF3, IFNαand IFNγ) and three pat-tern recognition receptors (TLR3, TLR4 and RIG1) was determined by qRT-PCR. The results showed that, most of the cyto-kines or receptors had obvious expression change compared with the control (without Poly I:C stimulation), especially the three cytokine genes IL6, IL8 and IL10, whose average expression change times were 20.71, 10.87 and 5.18, respectively. Ex-pression comparison between breeds and among individuals of the same breed indicated that there was obvious difference not only between Dapulain and Landrace (Dapulain higher than Landrace) but also among the three individuals of the same breed. Our study simulated the infection of dsRNA to host cells using Poly I:C, and provided experimental foundation for further study on selecting the immune genes in response to Poly I:C stimulation and identifying the unique disease-resistance genes of Dapulian.