【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1×10⁹ 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。更多还原

【Abstract】 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: 10 μg/capsule for 146S content of FMD O/rV-1 and A/rV-2 strains, and 1×10⁹ TCLD₅₀/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 0, 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-γ） 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-γ 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 three adjuvants significantly contributed to their elevation（P<0.05）, with the most significant elevation in the ISA 201 adjuvant group. In terms of CD4⁺/CD8⁺ ratio, all three adjuvant groups were significantly higher than the PBS control group（P<0.05）, and the ratio in the ISA 201 adjuvant group was significantly higher than those in the ISA 206 and SW adjuvant groups（P<0.05）. The results indicated that all three adjuvants could enhance the immunization effect of inactivated porcine foot-and-mouth disease-pseudorabies dual gene deletion vaccine to different degrees, with ISA 201 adjuvant having the best enhancement effect.更多还原