3种佐剂对猪口蹄疫-伪狂犬二联基因缺失灭活疫苗的免疫效果评价  

Evaluation of the immune effects of three adjuvants on inactivated foot-and-mouth disease-pseudorabies dual gene deletion vaccine in pigs

作　　者：王冬斌 吴涵 贾文青 李洪庆 白满达 张七斤[1] WANG Dongbin;WU Han;JIA Wenqing;LI Hongqing;BAI Manda;ZHANG Qijin(College of Veterinary Medicine,Inner Mongolia Agricultural University,Hohhot 010018,China;Biwei Antai Biotechnology Co.,Ltd,Hohhot 011500,China)

机构地区：[1]内蒙古农业大学兽医学院,呼和浩特010018 [2]必威安泰生物科技有限公司,呼和浩特011500

出　　处：《黑龙江畜牧兽医》2024年第11期95-100,共6页Heilongjiang Animal Science And veterinary Medicine

摘　　要：为了评价ISA 201、ISA 206和SW 3种佐剂对猪口蹄疫-伪狂犬二联基因缺失灭活疫苗的免疫增强效果,试验采用PCR方法鉴别口蹄疫O/rV-1株+A/rV-2株和伪狂犬RPVS1601-1株是否存在3B基因和gE基因缺失;将口蹄疫O/rV-1株+A/rV-2株和伪狂犬RPVS1601-1株抗原灭活并制成水相,分别与3种佐剂混合制成相应的灭活疫苗,使各抗原最终含量为口蹄疫O/rV-1株和A/rV-2株146S 10μg/头份,伪狂犬RPVS1601-1株1×10^(9)TCLD50/头份。将70只昆明小鼠随机分成ISA 201佐剂组(20只)、ISA 206佐剂组(20只)、SW佐剂组(20只)和PBS对照组(10只),肌肉注射制备的疫苗,首次免疫后2周进行二次免疫,首次免疫后第0,1,2,3,4,5,6周时进行断尾采血,收集血清,用ELISA方法检测白细胞介素-2(IL-2)、IL-4、γ干扰素(IFN-γ)含量和口蹄疫(O/rV-1型、A/rV-2型)抗体及伪狂犬抗体水平;并在首免2,4,6周时每组随机各取3只小鼠进行眼眶采血,用流式细胞仪检测CD4+和CD8+细胞百分比。结果表明:第3周时,ISA 201佐剂组的小鼠血清IL-2含量最高,显著高于ISA 206佐剂组和SW佐剂组(P<0.05);ISA 201佐剂组IFN-γ含量最高,三个佐剂组之间差异显著(P<0.05)。另外,所有佐剂组的IL-4含量在第2周开始显著高于PBS对照组(P<0.05)。第1~6周,ISA 201佐剂组抗体含量显著高于ISA 206佐剂组和SW佐剂组(P<0.05)。三个佐剂组CD4+/CD8+比值均显著高于PBS对照组(P<0.05),且ISA 201佐剂组的比值显著高于ISA 206佐剂组和SW佐剂组(P<0.05)。说明三种佐剂均能不同程度地提升猪口蹄疫-伪狂犬二联基因缺失灭活疫苗的免疫效果,其中ISA 201佐剂的提升效果最好。In order to evaluate the immune-enhancing effects of three adjuvants,ISA 201,ISA 206 and SW,on the inactivated porcine foot-and-mouth disease-pseudorabies dual gene deletion vaccine,in the experiment,PCR was used to identify the presence of 3B gene and gE gene deletion in FMD O/rV-1+A/rV-2 strain and pseudorabies RPVS1601-1 strain;and the antigens of FMD O/rV-1 strain+A/rV-2 strain and pseudorabies RPVS1601-1 strain were inactivated and made into aqueous phase,which were mixed with three adjuvants to make the corresponding inactivated vaccines,so that the final content of each antigen was:10μg/capsule for 146S content of FMD O/rV-1 and A/rV-2 strains,and 1×10^(9)TCLDso/head for pseudorabies RPVS1601-1 strain.Seventy Kunming mice were randomly divided into ISA 201 adjuvant group(20 mice),ISA 206 adjuvant group(20 mice),SW adjuvant group(20 mice)and PBS control group(10 mice);the prepared vaccine was injected intramuscularly,and the second immunization was performed 2 weeks after the first immunization.Blood was collected at O,1,2,3,4,5,and 6 weeks after the first immunization by tail-breaking;serum was collected;interleukin-2(IL-2),IL-4,and interferon gamma(IFN-)levels and foot-and-mouth disease(O/rV-1 type,A/rV-2 type)and pseudorabies antibodies were detected by ELISA;and three mice in each group were randomly selected for orbital blood collection at 2,4 and 6 weeks of the first immunization,and the percentages of CD4+and CD8+cells were detected by flow cytometry.The results showed that the highest serum levels of IL-2 were found in mice in the ISA 201 adjuvant group at the third week;the difference was significant(P<0.05)when compared with the ISA 206 adjuvant group and the SW adjuvant group;the highest levels of IFN-were found in the ISA 201 adjuvant group,and the difference between the three adjuvant groups was significant(P<0.05).In addition,IL-4 contents in all adjuvant groups started to increase significantly(P<0.05)in the second week compared to the PBS control group.For all three antibody contents,all t

关 键 词：猪口蹄疫-伪狂犬二联基因缺失灭活疫苗 佐剂 免疫效果 CD4+ CD8+ 细胞因子 

分 类 号：S855.3[农业科学—临床兽医学]