猪脾转移因子对La Sota株鸡新城疫弱毒疫苗的免疫增强作用
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福建省第七批引才“百人计划”;福建省科技计划项目(2019N0025,2021S2003);江苏省博士后科研资助计划(2019K072);福建省高层次人才和青年优秀人才访学研修资助计划(闽人社文〔2019〕239号);福州市科技计划项目(2020-PT-149);福建省畜禽疫病防控技术重大研发平台项目(2014N2003);福建省禽病防治重点实验室开放基金课题(KF201902);福建省家禽产业技术体系建设项目(2019-2022)


Immunoenhancement on Newcastle disease virus La Sota strain attenuated vaccine by pig spleen transfer factor
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    摘要:

    【目的】本文旨在研究猪脾转移因子(TF)对新城疫(ND)病毒弱毒疫苗La Sota株的免疫增强作用及机理,为兽医临床防控提供理论依据。【方法】以4种不同剂量La Sota疫苗株分别单独、与TF联合免疫SPF鸡,14 d后以ND病毒(NDV)参考强毒F48E9株(104.7 ELD50)进行攻毒,采用蛋白质芯片技术测定鸡外周血IL-6、IL-10、IL-16和IL-21浓度,并以血凝抑制(HI)试验和荧光定量RT-PCR方法分别检测鸡ND HI抗体效价和F48E9株的病毒血症水平。【结果】攻毒保护试验表明,单独免疫105.17、104.17、103.17和102.17 EID50剂量时疫苗攻毒保护率分别为100%、55%、0%和0%,半数保护量(PD50)为12 023 EID50;对应剂量联合免疫时疫苗攻毒保护率分别为100%、75%、0%和0%,PD50为6 918 EID50;对照组(非免疫有攻毒)和空白组(非免疫非攻毒)死亡率分别为100%和0%。对104.17 EID50免疫剂量进一步检测分析表明,免疫后,联合免疫鸡IL-6、IL-10、IL-21和ND HI抗体效价(log2x)比单独免疫最多时分别提高了254.95 pg/mL、62.10 pg/mL、1.51 pg/mL和2.6,其中ND HI抗体效价差异极显著(P<0.01);攻毒后,联合免疫鸡IL-10、IL-16、IL-21和病毒血症阳性率比单独免疫最多时分别降低了428.61 pg/mL、167.81 pg/mL、1.48 pg/mL和20%,其中IL-16、IL-21差异极显著(P<0.01)。【结论】本文结果显示,TF可提高La Sota株弱毒疫苗的免疫保护率及免疫后IL-6、IL-10、IL-21介导的免疫应答、ND HI抗体效价,降低疫苗PD50及攻毒后IL-16、IL-21介导的炎症反应、IL-10介导的免疫应答抑制作用、病毒血症。本文明确了TF对La Sota株弱毒疫苗具有较好的免疫增强作用,为疫苗免疫佐剂研发和ND防控提供参考。

    Abstract:

    [Objective] We evaluated the immunoenhancement effect and mechanism of pig spleen transfer factor (TF) on Newcastle disease virus (NDV) La Sota strain attenuated vaccine to provide a theoretical basis for the prevention and control of Newcastle disease (ND). [Methods] We inoculated La Sota strain alone and La Sota combined with TF into specific pathogen free (SPF) chickens at four different doses, and challenged SPF chickens by reference virulent NDV strain F48E9 (104.7 ELD50) 14 days post vaccination. In addition, we determined the concentrations of IL-6, IL-10, IL-16, and IL-21 in chicken peripheral blood by protein chip technology, the hemagglutination inhibition (HI) antibody titer by HI assay, and the viremia level of F48E9 by RT-PCR. [Results] The challenge test showed that the protection rates of vaccination with La Sota strain alone at the doses of 105.17, 104.17, 103.17, and 102.17 EID50 were 100%, 55%, 0%, and 0%, respectively, and the median protective dose (PD50) was 12 023 EID50. The protection rates of vaccination with La Sota strain combined with TF at the corresponding doses were 100%, 75%, 0%, and 0%, respectively, and the PD50 was 6 918 EID50. The mortality of control group (non-immunized but challenged) and blank group (non-immunized and non-challenged) were 100% and 0%. Upon vaccination at the dose of 104.17 EID50, the IL-6, IL-10, IL-21, and ND HI antibody titer (log2x) of the chicken exposed to combined vaccination increased at most by 254.95 pg/mL, 62.10 pg/mL, 1.51 pg/mL, and 2.6 (P<0.01), respectively, compared with those in the chicken vaccinated with La Sota strain alone. After challenge, the IL-10, IL-16, IL-21, and the viremia levels in the chicken under combined vaccination decreased at most by 428.61 pg/mL, 167.81 pg/mL (P<0.01), 1.48 pg/mL (P<0.01), and 20%, respectively, compared with those in the chicken vaccinated with La Sota strain alone. [Conclusion] TF improves the immune protection rate of NDV attenuated La Sota strain vaccine, the immune response mediated by IL-6, IL-10, and IL-21, and the ND HI antibody titer. In addition, TF reduces the PD50, the inflammatory responses mediated by IL-16 and IL-21, the inhibition of immune response mediated by IL-10, and the viremia after challenge. It is clear that TF has a good immunoenhancement effect on the La Sota strain attenuated vaccine, which will provide a reference for research and development of vaccine adjuvant as well as prevention and control of ND.

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徐磊,杨慧,刘毅发,杨子玚,闫丽萍,苏雅君,刘小龙,宋素泉,张渊魁,黄瑜. 猪脾转移因子对La Sota株鸡新城疫弱毒疫苗的免疫增强作用[J]. 微生物学报, 2022, 62(2): 727-741

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