3种免疫增强剂对猪伪狂犬灭活疫苗的免疫效果评价
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摘要
【目的】评价盐酸左旋咪唑(LH)、刀豆素(ConA)和胞壁酰二肽(MDP)3种免疫增强剂对猪伪狂犬病毒(PRV)灭活疫苗的免疫增强作用。【方法】将免疫增强剂混入灭活的PRV水相,和油相乳化后制成相应含有免疫增强剂的灭活疫苗,使各免疫增强剂终剂量为:LH 7.5 mg·mL~(–1)、ConA 25.0 mg·mL~(–1)、MDP 150.0 mg·mL~(–1)。首次免疫后14 d进行二次免疫,首次免疫后14 d、二次免疫后7和14 d采取小鼠血清,检测相关的细胞因子和特异性抗体水平,并在二次免疫后14 d攻毒,计算最终的攻毒保护率。【结果】相对于白油组,ConA组和MDP组在二次免疫后14 d对小鼠白介素2(IL-2)含量提升效果极显著;LH组在首次免疫后14 d、MDP组在二次免疫后14 d对白介素4(IL-4)含量提升效果显著;3种免疫增强剂分别在二次免疫后14和7 d对干扰素(IFN-γ)和特异性抗体含量提升效果极显著,并提高了攻毒保护率,LH组和ConA组存活率为20%,MDP组存活率为60%。【结论】3种免疫增强剂使用试验剂量均能不同程度地提升猪伪狂犬灭活疫苗的免疫效果,具有较好的应用前景。本研究结果为后续复合免疫增强剂筛选打下了基础。
【Objective】To evaluate the immunopotentiation effects of levamisole hydrochloride(LH),concanavalin A(ConA) and muramyl dipeptide(MDP) on pseudorabies virus(PRV) inactivated vaccine.【Method】The immunopotentiators were mixed into inactivated PRV virus aqueous phase, and emulsified with oil phase to make inactivated vaccines containing immunopotentiators. The final concentrations of immunopotentiators were LH 7.5 mg·mL~(–1), ConA 25.0 mg·mL~(–1) and MDP 150.0 mg·mL~(–1) respectively. The second immunization was on the 14 th day after the first immunization. Mouse serum samples were collected on the 14 th day after the first immunization, 7 th and 14 th day after the second immunization to detect relative cytokines(IL-2, IL-4, IFN-γ) and specific antibody levels. On the 14 th day after the second immunization, all mice were attacked and the final attack protective rate was calculated.【Result】Compared to Baiyou group,ConA and MDP groups extremely significantly improved IL-2 content in mouse serum on the 14 th day after thesecond immunization. LH group significantly improved IL-4 content on the 14 th day after the first immunization, and MDP group significantly improved IL-4 content on the 14 th day after the second immunization. The three immunopotentiators extremely significantly enhanced IFN-γ contents and specific antibody levels on the 14 th and 7 th day after the second immunization respectively, and also improved the attack protective rates. The survival rates of LH and ConA groups were 20%, while that of MDP group was 60%.【Conclusion】Three immunopotentiators within the range of the experimental dosages can enhance the immune effects of PRV inactivated vaccine in varying degrees.
【Objective】To evaluate the immunopotentiation effects of levamisole hydrochloride(LH),concanavalin A(ConA) and muramyl dipeptide(MDP) on pseudorabies virus(PRV) inactivated vaccine.【Method】The immunopotentiators were mixed into inactivated PRV virus aqueous phase, and emulsified with oil phase to make inactivated vaccines containing immunopotentiators. The final concentrations of immunopotentiators were LH 7.5 mg·mL~(–1), ConA 25.0 mg·mL~(–1) and MDP 150.0 mg·mL~(–1) respectively. The second immunization was on the 14 th day after the first immunization. Mouse serum samples were collected on the 14 th day after the first immunization, 7 th and 14 th day after the second immunization to detect relative cytokines(IL-2, IL-4, IFN-γ) and specific antibody levels. On the 14 th day after the second immunization, all mice were attacked and the final attack protective rate was calculated.【Result】Compared to Baiyou group,ConA and MDP groups extremely significantly improved IL-2 content in mouse serum on the 14 th day after thesecond immunization. LH group significantly improved IL-4 content on the 14 th day after the first immunization, and MDP group significantly improved IL-4 content on the 14 th day after the second immunization. The three immunopotentiators extremely significantly enhanced IFN-γ contents and specific antibody levels on the 14 th and 7 th day after the second immunization respectively, and also improved the attack protective rates. The survival rates of LH and ConA groups were 20%, while that of MDP group was 60%.【Conclusion】Three immunopotentiators within the range of the experimental dosages can enhance the immune effects of PRV inactivated vaccine in varying degrees.
引文
[1]AKIRA S,TAKEDA K,KAISHO T.Toll-like receptors:Critical proteins linking innate and acquired immunity[J].Nat Immunol,2001,2(8):675-680.
[2]KRISTENSEN C S,ANDREASEN M,ERSB?LL A K,et al.Antibody response in sows and piglets following vaccination against Mycoplasma hyopneumoniae,toxigenic Pasteurella multocida,and Actinobacillus pleuropneumoniae[J].Can J Vet Res,2004,68(1):66-70.
[3]FISCHER T,BüTTNER M,RZIHA H J.T helper 1-type cytokine transcription in peripheral blood mononuclear cells of pseudorabies virus(Suid herpesvirus 1)-primed swine indicates efficient immunization[J].Immunology,2000,101(3):378-387.
[4]李向东,王莹,苏培,等.盐酸左旋咪唑栓免疫调节作用的临床观察[J].天津药学,2000,12(2):44-45.
[5]牛华星,李淑花,张传津,等.盐酸左旋咪唑-维生素E可溶性粉对鸡免疫功能增强作用研究[J].山东畜牧兽医,2009,30(8):3-5.
[6]陈华良.一种动物复合免疫增强剂的研制[D].广州:华南农业大学,2016.
[7]翁梅悦,银少华,欧盛绘,等.猪场常见重要病毒性疫病临床诊断与防控概要[J].畜牧兽医,2017,326(2):90-92.
[8]YOO Y C,YOSHIMATSU K,KOIKE Y,et al.Adjuvant activity of muramyl dipeptide derivatives to enhance immunogenicity of a hantavirus-inactivated vaccine[J].Vaccine,1998,16(2/3):216-224.
[9]田世昭.分枝杆菌的佐剂活性与菌株[J].国外医学(生物制品学分册),1982(1):23.
[10]陈华良,王和兴,李小鹏,等.广东地区猪伪狂犬病流行病学调查与g E、TK基因遗传进化分析[J].广东畜牧兽医科技,2016,41(2):5-9.
[11]裴春生,何玉友,李群,等.油乳剂疫苗中硬脂酸铝含量与油相水相比例关系研究[J].中国兽药杂志,2005,39(2):21-23.
[12]TANG Y,LU J,WU P,et al.Inactivated vaccine with adjuvants consisting of pattern recognition receptor agonists confers protection against avian influenza viruses in chickens[J].Vet Microbiol,2014,172(1/2):120-128.
[13]司兴奎,郭鑫,杨汉春.猪圆环病毒2型感染后猪外周血淋巴细胞IL-2、IL-4、IL-10、IL-12p40、IFN-γ和TNF-αm RNA转录的变化[J].畜牧兽医学报,2009,40(1):78-82.
[2]KRISTENSEN C S,ANDREASEN M,ERSB?LL A K,et al.Antibody response in sows and piglets following vaccination against Mycoplasma hyopneumoniae,toxigenic Pasteurella multocida,and Actinobacillus pleuropneumoniae[J].Can J Vet Res,2004,68(1):66-70.
[3]FISCHER T,BüTTNER M,RZIHA H J.T helper 1-type cytokine transcription in peripheral blood mononuclear cells of pseudorabies virus(Suid herpesvirus 1)-primed swine indicates efficient immunization[J].Immunology,2000,101(3):378-387.
[4]李向东,王莹,苏培,等.盐酸左旋咪唑栓免疫调节作用的临床观察[J].天津药学,2000,12(2):44-45.
[5]牛华星,李淑花,张传津,等.盐酸左旋咪唑-维生素E可溶性粉对鸡免疫功能增强作用研究[J].山东畜牧兽医,2009,30(8):3-5.
[6]陈华良.一种动物复合免疫增强剂的研制[D].广州:华南农业大学,2016.
[7]翁梅悦,银少华,欧盛绘,等.猪场常见重要病毒性疫病临床诊断与防控概要[J].畜牧兽医,2017,326(2):90-92.
[8]YOO Y C,YOSHIMATSU K,KOIKE Y,et al.Adjuvant activity of muramyl dipeptide derivatives to enhance immunogenicity of a hantavirus-inactivated vaccine[J].Vaccine,1998,16(2/3):216-224.
[9]田世昭.分枝杆菌的佐剂活性与菌株[J].国外医学(生物制品学分册),1982(1):23.
[10]陈华良,王和兴,李小鹏,等.广东地区猪伪狂犬病流行病学调查与g E、TK基因遗传进化分析[J].广东畜牧兽医科技,2016,41(2):5-9.
[11]裴春生,何玉友,李群,等.油乳剂疫苗中硬脂酸铝含量与油相水相比例关系研究[J].中国兽药杂志,2005,39(2):21-23.
[12]TANG Y,LU J,WU P,et al.Inactivated vaccine with adjuvants consisting of pattern recognition receptor agonists confers protection against avian influenza viruses in chickens[J].Vet Microbiol,2014,172(1/2):120-128.
[13]司兴奎,郭鑫,杨汉春.猪圆环病毒2型感染后猪外周血淋巴细胞IL-2、IL-4、IL-10、IL-12p40、IFN-γ和TNF-αm RNA转录的变化[J].畜牧兽医学报,2009,40(1):78-82.