通过免疫控制口蹄疫 (1)

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1、FOOT AND MOUTH DISEASE CONTROL BY VACCINATION 通过免疫控制口蹄疫 Dr.Shih-Ping Chen 陈世平博士 Agriculture Technology Research Institute 农业科技研究院 Agricultural Technology Research Institute Outlines 纲要纲要 FMD in Taiwan 台湾的口台湾的口蹄蹄疫疫 Serum neutralizing titers and protection 血清中血清中和抗体滴和抗体滴度度和保护和保护 Vaccine releasing test

2、 in Taiwan 台湾疫苗上市试验台湾疫苗上市试验 Field trials of vaccinated pigs 免疫猪的生产试验免疫猪的生产试验 Selection of right vaccine strain 选择合适的疫苗选择合适的疫苗株株 Duration of Immunity and Vaccination schedule 免免疫持续时疫持续时间和免疫程序间和免疫程序 Discussion 讨论讨论 Agricultural Technology Research Institute FMD in Taiwan 台台湾的口蹄疫湾的口蹄疫 In Taiwan 97,a sp

3、ecies-specific adaptation to pigs 1997年年台湾地区台湾地区，种种特异特异性表现在猪上性表现在猪上 Overt clinical signs in pigs 猪有明猪有明显的临床症状显的临床症状 Vaccination has been successfully used for controlling FMD 免疫已免疫已成功用于控制口蹄疫成功用于控制口蹄疫 Been recognized as FMD free with vaccination before 2008 在在2008年之年之前由于免疫被前由于免疫被确认为无口蹄确认为无口蹄疫疫 Effect

4、ive methods of differentiating vaccinated pigs from infected 鉴别免疫猪和被鉴别免疫猪和被感感染猪染猪的有效方法的有效方法 Agricultural Technology Research Institute Agricultural Technology Research Institute Humoral responses are more important than the cytotoxic reactions in protective immune response against FMD 对口蹄疫保护性免疫反应对口蹄

5、疫保护性免疫反应，体液免疫反应比细体液免疫反应比细胞毒性反应更重要胞毒性反应更重要 FMD SN titers do correlate with the protection in cattle 口蹄口蹄疫血清中和滴疫血清中和滴度度与在牛身上的保护作与在牛身上的保护作用相关用相关 Determine the protected SN titer in pigs 确确定猪定猪的保护的血清中和抗体滴度的保护的血清中和抗体滴度 Relationship between serum neutralizing titers and protection 血清中血清中和滴和滴度和保度和保护力之护力之间的

6、关系间的关系 Agricultural Technology Research Institute Relationship between serum neutralizing titers and protection from FMD in pigs after oil emulsion vaccination 油乳剂疫苗接种后的血清中油乳剂疫苗接种后的血清中和抗体滴和抗体滴度度和和对口蹄疫保护之间的对口蹄疫保护之间的关系关系 Black L et al.1984.血清抗体滴度（log 10)Agricultural Technology Research Institute Corre

7、lations Between Serum Neutralization Antibody and Protection of Foot-and-Mouth Disease In Pigs Lin et al.2004.血血清中和抗清中和抗体和对猪的口蹄体和对猪的口蹄保护的相关性保护的相关性 248 pigs were vaccinated with double oil emulsion vaccines of by IM once at 8-week old(74 pigs)challenged at 4 weeks post vaccination or twice at 8-and 1

8、2-week old(174 pigs)challenged at 20 WPV 248头猪用双油乳剂疫苗在第头猪用双油乳剂疫苗在第8周肌肉注射免疫一次周肌肉注射免疫一次（74头头），免疫后免疫后4周攻毒周攻毒，或第或第8、12 周免疫两次周免疫两次，免疫后免疫后20周攻毒周攻毒 Homogeneous strain was used for challenge(O/TAW strain)用同源毒株攻毒用同源毒株攻毒（O/TAW 株株）Subcutaneous injection with 105 TCID50 of FMDV into the heel bulbs of the right

9、forefoot 在右前脚根部用在右前脚根部用105 TCID50 口蹄疫病毒皮下注射口蹄疫病毒皮下注射 Agricultural Technology Research Institute Correlation between SN antibody titers and protection against FMD virus challenge.Pigs in panel A were challenged at 4 WPV and in panel B were challenged at 20 WPV.Filled circles,protection;Open circles,n

10、o protection.血清中和抗血清中和抗体滴度体滴度和对攻毒保护的和对攻毒保护的相关性相关性。A组组 免疫后免疫后4周攻毒周攻毒，B组组 免疫免疫后后20周攻毒周攻毒。实心圆实心圆，保护保护;空空心圆心圆，没没有保护有保护。Agricultural Technology Research Institute SN titers did not correlate well with the protection,but pigs with SN titers 1:64 did provide a better protection for pigs of the 4 WPV。血清中和抗体

11、滴度与保护力没有好血清中和抗体滴度与保护力没有好的相的相关性关性，但中和抗体滴度但中和抗体滴度1:64的猪对的猪对4周周后攻毒的猪后攻毒的猪提供提供了较好的了较好的保保护护。Agricultural Technology Research Institute Batch tests in swine with FMD vaccines:Viral Neutralization Test for batch releasing test 猪口蹄疫疫苗批次实验：批次上市的病毒中和试验 Agricultural Technology Research Institute Vaccine releas

12、ing test in Taiwan 台湾的疫苗上市试验台湾的疫苗上市试验 FMD vaccine quality is critical for the control of FMD and in Taiwan this quality was evaluated by SN titers instead of by the challenge test 口蹄疫疫苗质量对口蹄疫控制至关重要口蹄疫疫苗质量对口蹄疫控制至关重要，在台湾在台湾，质量是通过中和抗体滴定来评估质量是通过中和抗体滴定来评估，而不是攻毒试验而不是攻毒试验 In vaccine test,the geometric mean

13、 SN antibody titers of 5 pigs were determined after 3 or 4 weeks of vaccination and vaccines producing titers 1:32 was considered to pass the test 在疫苗试验中在疫苗试验中，5头猪头猪的血清中和抗体的几的血清中和抗体的几何平何平均值在免疫后均值在免疫后3或或4周来评估周来评估，达到滴达到滴度度1:32 的疫苗为的疫苗为通过测试通过测试 Agricultural Technology Research Institute Duration of imm

14、unity and MDA interference trials:different swine vaccination studies in Taiwan after 1997 FMD outbreaks 免疫持续时间和MDA干扰实验：1997年口蹄疫疫情爆发后台湾不同的免疫研究 Agricultural Technology Research Institute Vaccination study after 1997 outbreak of FMD 1997爆发口蹄疫后的免疫研究爆发口蹄疫后的免疫研究 Seventy pigs,8-week-old,on farm L were ran

15、domly allocated into one of seven groups L场场70 头头8周周龄的猪随机龄的猪随机分为七分为七组组 Vaccinated with one of seven commercially available monovalent FMD serotype O1 Manisa,O 4174 or O1 Campos oil-emulsified vaccines(Vaccines AHsee Table 1)用用7种单价油乳剂商品苗种单价油乳剂商品苗，口蹄疫血清型口蹄疫血清型O1马尼萨马尼萨，O4174或或O1坎波斯免疫坎波斯免疫（疫苗疫苗A-H-见表见表1

16、）All pigs received the first FMD vaccination at 8 weeks of age and a secondary vaccination at 12 weeks of age 所有所有猪在第猪在第8周龄免疫第一针周龄免疫第一针，12周周龄第二针龄第二针 Serum samples were collected from pigs at 8,12,16 and 23 weeks of age 第第8，12，16和和23周周龄采集血清样本龄采集血清样本 Measurement of neutralizing antibody titers and NSP

17、 antibodies.测定测定中和抗体效价中和抗体效价和和NSP抗抗体体。Agricultural Technology Research Institute Seven commercially available FMD vaccines used in the vaccination experiments 在免疫实在免疫实验中使用的验中使用的七种商品口七种商品口蹄疫疫苗蹄疫疫苗 Vaccine designation Origin of vaccine Virus strain A Argentina O1 Campos B Argentina O1 Campos C Argenti

18、na O1 Campos D England O 4174 E Germany O1 Manisa F Turkey O1 Manisa G Holland O1 Manisa 疫苗编号疫苗编号 疫苗来源疫苗来源 毒株毒株 Agricultural Technology Research Institute 012345678901234561115Titerweeks post vaccinationTwice vaccinationABCDEFGPrimary vaccination Boost 两次疫苗接种两次疫苗接种 滴滴度度 初初级免疫级免疫 加强加强 接种后周数接种后周数 Agri

19、cultural Technology Research Institute 0123456701234561115Mean Log2 TiterWeeks post-vaccinationSingle vaccinationABCDEFG单次免疫单次免疫 平均Log2滴度 免疫后免疫后周周数数 Agricultural Technology Research Institute NSP 抑制抑制%阳性阳性%疫苗 第一次免疫前 第一次免疫前 第一次免疫后15周 第一次免疫后15周 Agricultural Technology Research Institute Appropriate va

20、ccine strains related to OTaiwan/97 were determined based on the evaluation of r1 values 根据评估根据评估r1值确定与值确定与OTaiwan/97相关的免疫毒株相关的免疫毒株 The r1 values for O1 Manisa,O 4174 and O1 Campos were 1,1 and 0.7,respectively，O1 Manisa,O 4174 和和 O1 Campos 的的r1值分别为值分别为 1,1 和和 0.7,However,vaccines E,F and G formulat

21、ed with O1 Manisa induced lower neutralizing titers than those vaccines with O1 Campos and O 4174.然而然而，用用O1 Manisa 生产的生产的E、F、和和G 比用比用O1 Campos 和和 O 4174生产的疫苗产生较低的中和抗生产的疫苗产生较低的中和抗体滴度体滴度 Other factors which can cause a difference in vaccine efficacy include antigen mass contained in vaccines,vaccine p

22、roduction procedures and antigen adjuvant interactions其其它导致疫苗效果差异的因素包括疫苗中抗原量它导致疫苗效果差异的因素包括疫苗中抗原量，疫苗生产疫苗生产过程及抗原与佐剂的相互作用过程及抗原与佐剂的相互作用 Agricultural Technology Research Institute 050100150200TiterMohthly Monitoring Monitoring titers of sows after vaccination 免疫后监免疫后监测母测母猪体中滴猪体中滴度度 滴度 每月监控每月监控 Agricultur

23、al Technology Research Institute Immune response of sows after vaccination with heterologous strains 母猪接种异源毒株后的免疫反应母猪接种异源毒株后的免疫反应 免疫时间免疫时间 中和滴度中和滴度 Agricultural Technology Research Institute Comparison of neutralizing titers in pigs vaccinated with homologous and heterologous vaccines 比较用同比较用同源和异源和异

24、源毒株疫苗免疫的中和抗体滴源毒株疫苗免疫的中和抗体滴度度 Agricultural Technology Research Institute 中和滴度 免疫时间免疫时间 Agricultural Technology Research Institute 中和滴度 免疫时间免疫时间 Agricultural Technology Research Institute 中和滴度 免疫时间免疫时间 Agricultural Technology Research Institute Effect of maternal Ab on the antibody titers of pigs at 4

25、 weeks after vaccination 免疫4周后母 抗对猪的抗体滴度的作用 Agricultural Technology Research Institute 仔豬移行抗體仔豬移行抗體 log10 Matermal Ab 母抗母抗 Agricultural Technology Research Institute Matermal Ab 母抗母抗 Agricultural Technology Research Institute 仔豬移行抗體仔豬移行抗體 log10 Matermal Ab 母抗母抗 168 pigs free from FMD randomly divide

26、d into three groups 168头没有口蹄疫的猪被随机分头没有口蹄疫的猪被随机分成三组成三组 A-imported from England A组组-从英国进口从英国进口 B-imported from Argentina B组组-从阿根廷进口从阿根廷进口 C-imported from Russia C组组-从俄罗斯进口从俄罗斯进口 Field trials 生产试验 Agricultural Technology Research Institute 第一次免疫后周数第一次免疫后周数 中中和和滴滴度度 Agricultural Technology Research Inst

27、itute For all three vaccines,geometric mean neutralizing antibody titers peaked by 2 weeks after booster vaccination and had declined by 12 weeks post vaccination but were still higher at that time than titers 4 weeks after primary vaccination.对于对于所有三种疫苗所有三种疫苗，中中和抗体滴和抗体滴度几何平均度几何平均值值加加强强免疫免疫2周周后达到最高后

28、达到最高，免疫免疫12周周后下降后下降，但仍高但仍高于于初免初免4周周后后的滴度的滴度。Profile of Neutralizing Titer with Double doses 两针免疫中和抗体滴度的情况两针免疫中和抗体滴度的情况 第一次接种第一次接种后周数后周数 中和滴度 Agricultural Technology Research Institute Sequential neutralizing antibody titers in pigs vaccinated with double dose of vaccines B and C at 12,16 and 20 week

29、s of age and tested for antibodies at 12,16,20 and 24 weeks of age.在在12，16和和20周龄猪中接种周龄猪中接种两针两针疫苗疫苗B和和C的的连续连续中和抗体滴度中和抗体滴度，检测检测在在12，16，20和和24周龄的周龄的抗体抗体.For both vaccines geometric mean neutralizing antibody titers increased 4 weeks after each vaccination.对于这两种疫对于这两种疫苗苗，中中和抗体滴和抗体滴度几度几何平均何平均值值每每次接种次接种后后

30、4周周 增加增加.Agricultural Technology Research Institute Selection of right vaccine strain 选择合适的疫苗选择合适的疫苗株株 Antigenic diversity(serotypes)抗原多样性抗原多样性（血清型血清型）Select vaccine strains from actual field isolates or traditional vaccine strains?选选择择猪场分离出的毒株猪场分离出的毒株或或传统疫苗传统疫苗株株?FMD naive country:口蹄疫阴性的国家口蹄疫阴性的国家

31、seronegative pigs:血血清阴清阴性猪性猪：Single vaccination of seronegative pigs protects 3-6 months(data depending on vaccine strains)血清阴性猪血清阴性猪单次接种可保单次接种可保护护3 3-6 6个月个月（根根据疫苗据疫苗株株的数据的数据）Sow vaccination with Heterologous/homologous vaccine strains 母猪接种异源母猪接种异源/同源疫同源疫苗株苗株 Heterologous vaccine strains:3 months 异

32、源的疫苗株：异源的疫苗株：3个月个月 Homologous vaccine strain:6 months 同同源疫苗株：源疫苗株：6个月个月 Duration of Immunity and Vaccination schedule 免疫免疫持续时间免疫程序持续时间免疫程序 Agricultural Technology Research Institute FMD infected/vaccinated country:口蹄疫病毒感口蹄疫病毒感染染/免疫免疫的的国家：国家：seropositive pigs:血清阳性猪：血清阳性猪：Multiple vaccinated sows with

33、 natural infection have high antibody titers自然感染自然感染并并多多次免疫的次免疫的母猪有母猪有较高的抗体滴度较高的抗体滴度 Variable MDA titers in piglets 仔猪中仔猪中MDA滴滴度度 有差异有差异 Interference of heterologous vaccines with high MDA titers有高有高MDA滴度的异滴度的异源疫源疫苗的干苗的干扰扰 Two vaccination in fatteners:育肥猪两次免疫育肥猪两次免疫：1st vaccination at 8 weeks and 2nd vaccination at 12 weeks with homologous vaccine 用同源毒株疫苗在用同源毒株疫苗在8周周和和12周周两次免疫两次免疫 Delay of vaccination time when using heterologous vaccines 用用异异源源毒株毒株疫疫苗时苗时，推迟免疫推迟免疫时间时间 Discussion 讨论讨论