猪链球菌2型灭活疫苗对小鼠的免疫效果评价

doi:10.3969/j.issn.1004-1524.2020.04.03

浙江农业学报 ›› 2020, Vol. 32 ›› Issue (4): 577-585.DOI: 10.3969/j.issn.1004-1524.2020.04.03

猪链球菌2型灭活疫苗对小鼠的免疫效果评价

段倩倩, 毛天骄, 韩业芹, 韩雪姣, 魏建忠, 孙裴, 李郁^*

安徽农业大学动物科技学院,安徽合肥 230036

收稿日期:2019-09-14 出版日期:2020-04-25 发布日期:2020-04-26
通讯作者: *李郁,E-mail: liyouer@163.com
作者简介:段倩倩(1996—),女,安徽合肥人,硕士研究生,研究方向为动物传染病学。E-mail: 1460809869@qq.com
基金资助:
国家星火计划重点项目(2014GA710002); 安徽省长三角联合科技攻关项目(1101c0603065); 安徽省质量工程项目(2013sxzx008); 安徽省生猪产业体系基金(皖农科〔2016〕84号)

Evaluation of immune efficacy of inactivated Streptococcus suis serotype 2 vaccine in mice

DUAN Qianqian, MAO Tianjiao, HAN Yeqin, HAN Xuejiao, WEI Jianzhong, SUN Pei, LI Yu^*

College of Animal Science and Technology, Anhui Agricultural University, Hefei 230036, China

Received:2019-09-14 Online:2020-04-25 Published:2020-04-26

摘要/Abstract

摘要： 在前期筛选出2株毒力强、抗原性好、遗传稳定的猪链球菌2型(SS2)疫苗菌株(HF2、HF3株)并分别制成灭活疫苗(采用ISA 201 VG矿物油佐剂)的基础上,进一步与SS2商品化灭活疫苗(HA9801株,铝胶佐剂)同步免疫小鼠,利用间接ELISA、流式细胞术、免疫攻毒试验、细菌定植试验和病理组织学观察等方法测定小鼠血清中IgG抗体效价,细胞因子含量(IL-4、IL-10、IFN-γ、TNF-β、MCP-1),外周血中CD4⁺/CD3⁺、CD8⁺/CD3⁺ T细胞亚群含量,攻毒保护率,组织荷菌数和病理组织变化等指标。结果显示,二免7 d后,HF2、HF3灭活疫苗组和HA9801商品化灭活疫苗组的血清IgG抗体效价分别为1:25 600、1:12 800、1:25 600;HF2灭活疫苗组的IL-4、IL-10含量显著高于HF3灭活疫苗组和HA9801商品化灭活疫苗组,IFN-γ、TNF-β含量显著高于HF3灭活疫苗组,但MCP-1含量显著低于HF3灭活疫苗组;HF2和HF3灭活疫苗组的CD4⁺/CD3⁺ T细胞比率均显著低于HA9801商品化灭活疫苗组,但CD8⁺/CD3⁺ T细胞比率差异不显著;HF2、HF3灭活疫苗和HA9801商品化灭活疫苗对小鼠的攻毒保护率均为100%;HF2灭活疫苗组小鼠的肺、脾脏组织荷菌数显著低于HF3灭活疫苗组;HF2灭活疫苗组小鼠的肺、肾、脾脏病理变化较HF3灭活疫苗组和HA9801商品化灭活疫苗组轻微。综合结果表明,由SS2(HF2株)制备的ISA 201 VG佐剂灭活疫苗免疫小鼠后不仅可产生较强的免疫应答,还可完全抵抗强毒株的攻击。

关键词: 猪链球菌2型, 灭活疫苗, 免疫效果, 小鼠

Abstract: In preliminary research, 2 vaccine strains of Streptococcus suis serotype 2 HF2, HF3 were screened out through pathogenicity test, antigenicity test and stability test, and been prepared into inactivated vaccine with ISA 201 VG mineral oil adjuvant. These two inactivated vaccines were immunized in mice with commercial vaccine to evaluate their immune efficacy. The titer of IgG antibody and content of cytokines (IL-4, IL-10, IFN-γ, TNF-β, MCP-1) in serum were detected by ELISA and the subpopulation of T cells of CD4⁺/CD3⁺, CD8⁺/CD3⁺ were measured by flow cytometry. The protective rate of each immunization group after challenge test, the colonization of SS2 in mice by tissue load test and the pathological sections of lung, liver, spleen and kidney of mice were collected and observed. The results showed that the serum IgG antibody titers of the SS2 inactivated vaccine (HF2 strain, HF3 strain) and the SS2 commercial inactivated vaccine (HA9801 strain) after the second immunization of mice were 1:25 600, 1:12 800, 1:25 600, respectively. The content of IL-4 and IL-10 induced by HF2 inactivated vaccine were significantly higher than HF3 inactivated vaccine and commercial vaccine. The content of IFN-γ、TNF-β were significantly higher than HF3 inactivated vaccine, and the content of MCP-1 were significantly lower than HF3 inactivated vaccine. The commercial vaccine group induced mice to produce significantly higher ratio of CD4⁺/CD3⁺ T cells than other groups. The protection rate of HF2, HF3 inactivated vaccine and HA9801 commercial inactivated vaccine on mice were all 100%.The bacteria colonization of lung and spleen of HF2 inactivated vaccine were significantly lower than HF3 inactivated vaccine, and the pathological changes of lung, spleen and kidney were lighter than other groups. In conclusion, HF2 inactivated vaccine can not only produce a higher level of humoral immunity and cellular immunity, but also completely resist the attack of the virulent strain.

Key words: Streptococcus suis serotype 2, inactivated vaccine, immune efficacy, mice

中图分类号:

S855.3

段倩倩, 毛天骄, 韩业芹, 韩雪姣, 魏建忠, 孙裴, 李郁. 猪链球菌2型灭活疫苗对小鼠的免疫效果评价[J]. 浙江农业学报, 2020, 32(4): 577-585.

DUAN Qianqian, MAO Tianjiao, HAN Yeqin, HAN Xuejiao, WEI Jianzhong, SUN Pei, LI Yu. Evaluation of immune efficacy of inactivated Streptococcus suis serotype 2 vaccine in mice[J]. , 2020, 32(4): 577-585.

参考文献

[1] 金梅林. 猪链球菌致病分子基础与防控技术[M]. 北京: 科学出版社, 2018: 6-7.
[2] GOTTSCHALK M, SEGURA M.The pathogenesis of the meningitis caused by Streptococcus suis: the unresolved questions[J]. Veterinary Microbiology, 2000, 76(3): 259-272.
[3] 罗满林. 兽医生物制品学[M]. 北京: 中国农业大学出版社, 2019: 12-23.
[4] 陈章, 吴华健, 毛天骄, 等. 猪链球菌2型制苗用菌株的筛选[J]. 浙江农业学报, 2020, 32(1): 57-64.
CHEN Z, WU H J, MAO T J, et al.Screening of strain for inactivated vaccine of Streptococcus suis type 2[J]. Acta Agriculturae Zhejiangensis, 2020, 32(1): 57-64.(in Chinese with English abstract)
[5] 杜德超, 于岩飞, 马彩凤, 等. 猪链球菌2型不同mrp基因型与毒力的关系[J]. 微生物学通报, 2016, 43(1): 147-155.
DU D C, YU Y F, MA C F, et al.Genotype of mrp and its relationship with the virulence of Streptococcus suis serotype 2[J]. Microbiology China, 2016, 43(1): 147-155.(in Chinese with English abstract)
[6] 吴华健, 毛天骄, 杨龙斌, 等. 猪链球菌2型溶血素基因的表达及其免疫原性[J]. 微生物学通报, 2019, 46(3): 624-637.
WU H J, MAO T J, YANG L B, et al.Expression and immunogenicity of the sly gene of Streptococcus suis type 2[J]. Microbiology China, 2019, 46(3): 624-637.(in Chinese with English abstract)
[7] 李丽, 黄良宗, 谢博, 等. 猪链球菌2型疫苗候选菌株的筛选[J]. 黑龙江畜牧兽医, 2018(17): 160-163.
LI L, HUANG L Z, XIE B, et al.Screening of candidate strains of Streptococcus suis type 2 vaccine[J]. Heilongjiang Animal Science and Veterinary Medicine, 2018(17): 160-163.(in Chinese)
[8] 薛云, 李静, 王臣, 等. 羊源致病性柠檬酸杆菌4种佐剂灭活疫苗的免疫效果比较[J]. 中国预防兽医学报, 2018, 40(8): 726-731.
XUE Y, LI J, WANG C, et al.Comparison of four kinds of adjuvants for inactivated Citrobacter freundii vaccine in sheep[J]. Chinese Journal of Preventive Veterinary Medicine, 2018, 40(8): 726-731.(in Chinese with English abstract)
[9] 刘帅. 两种佐剂对虹鳟杀鲑气单胞菌灭活疫苗免疫效果的影响[D]. 上海: 上海海洋大学, 2017.
LIU S.Immune effect of two adjuvants on rainbow trout (Oncorhynchus mykiss) immunized by inactivated Aeromonas salmonicida vaccine[D]. Shanghai: Shanghai Ocean University, 2017. (in Chinese with English abstract)
[10] 张超范, 崔尚金, 苗岚飞, 等. 猪细小病毒灭活疫苗安全性试验及佐剂的筛选[J]. 中国预防兽医学报, 2009, 31(6): 462-465.
ZHANG C F, CUI S J, MIAO L F, et al.Safety valuation of a porcine parvovirus inactivated vaccine[J]. Chinese Journal of Preventive Veterinary Medicine, 2009, 31(6): 462-465.(in Chinese with English abstract)
[11] CHEN, TANG J Q, DONG W, et al. A glimpse of streptococcal toxic shock syndrome from comparative genomics of S. suis 2 Chinese isolates[J]. PLoS One, 2007, 2(3): e315.
[12] FAN H Y, PAN Y F, FANG L R, et al.Construction and immunogenicity of recombinant pseudotype baculovirus expressing the capsid protein of porcine Circovirus type 2 in mice[J]. Journal of Virological Methods, 2008, 150(1/2): 21-26.
[13] 魏子贡. 猪链球菌流行病学及其生物被膜形成机理研究[D]. 武汉: 华中农业大学, 2010.
WEI Z G.Study on the epidemiology of Streptococcus suis and the merchanism of their biofilm formation[D]. Wuhan: Huazhong Agricultural University, 2010.(in Chinese)
[14] DAR P, KALAIVANAN R, SIED N, et al.Montanide ISA^TM 201 adjuvanted FMD vaccine induces improved immune responses and protection in cattle[J]. Vaccine, 2013, 31(33): 3327-3332.
[15] MOSMANN T R, CHERWINSKI H, CHER D, et al.Two types of mouse helper T cell clone: differences in B cell help and lymphokine synthesis[M]//Molecular Basis of Lymphokine Action. Totowa, NJ: Humana Press, 1987: 149-159.
[16] ZUCKERMANN F A, GASKINS H R.Distribution of porcine CD4/CD8 double-positive T lymphocytes in mucosa-associated lymphoid tissues[J]. Immunology, 1996, 87(3): 493-499.

猪链球菌2型灭活疫苗对小鼠的免疫效果评价

Evaluation of immune efficacy of inactivated Streptococcus suis serotype 2 vaccine in mice

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