不同组合猪繁殖与呼吸综合征疫苗对仔猪的免疫效果研究

江兴华

doi:10.19303/j.issn.1008-0384.2022.009.002

不同组合猪繁殖与呼吸综合征疫苗对仔猪的免疫效果研究

doi: 10.19303/j.issn.1008-0384.2022.009.002

江兴华

兆丰华生物科技（福州）有限公司，福建　福州 350014

基金项目: 福州市科技创新平台项目（2021-P-036）；福建省科技重大专项（2021NZ029008）

详细信息

作者简介:
江兴华（1975−），男，高级兽医师，研究方向：兽用生物制品的生产、管理与研发（E-mail：jxhgreen@163.com）

中图分类号: S 852
计量
- 文章访问数: 333
- HTML全文浏览量: 86
- PDF下载量: 41
- 被引次数: 0
出版历程
- 收稿日期: 2022-08-02
- 修回日期: 2022-09-02
- 网络出版日期: 2022-10-26
- 刊出日期: 2022-09-30

Immune Effect of Different Vaccine Combinations of Porcine Reproductive and Respiratory Syndrome On Piglets

JIANG Xinghua

JOFUNHWA Biotechnology (Fuzhou) Co., Ltd., Fuzhou, Fujian 350014, China

摘要

摘要: 目的设计猪繁殖与呼吸综合征PRRS活疫苗与灭活疫苗4种不同组合方式免疫仔猪，最大限度地发挥其对仔猪的免疫效果，为生产上制定PRRS的疫苗防控方案提供参考。方法制定4组不同组合的PRRS疫苗免疫方案，筛选 25头28~35 d仔猪，随机分成5组。A组为使用PRRS活疫苗（CH-1R株）A进行一免，每头注射2 mL（含1头份），间隔14 d使用PRRS灭活疫苗（CH-1a株）A进行二免，每头肌肉注射2 mL；B组免疫同A组，只是一免用PRRS活疫苗（CH-1R株）B，二免用PRRS灭活疫苗（CH-1a株）B；C组为一免与二免均使用PRRS灭活疫苗（CH-1a株）A，两次免疫间隔14 d，每次肌肉注射2 mL；D组为一免与二免均使用PRRS灭活疫苗（CH-1a株）B，其他方法同C组；E组为未免对照组。各组均于免疫前、一免后1、3、5、7 d进行采外周血检测细胞因子含量（IL-2、IFN-γ、IFN-α、TNF-α），于一免前、一免后14 d、二免后7、14、21、28 d进行采血检测免疫后抗体水平（GP5蛋白与N蛋白抗体），各组于二免后28 d进行攻毒，攻毒后21 d内观察体温变化，21d后进行剖检观察肺脏的病变。结果各免疫组一免后细胞因子含量发生不同的变化，对照组IL-2随日龄的增大有明显的下降，A、B组在3 d时显著高于其他免疫组；C、D组的IFN-γ含量在3、5、7 d时均显著高于A、B组；B组的 IFN-α含量在一免后3、5、7 d时显著高于其他组，A组的TNF-α含量在7 d时显著高于其他组。攻毒后对照组的体温上升快且高，高温持续期长，B组温度上升幅度小，较快降至常温。免后不同时间抗体水平、抗体阳性率、攻毒保护率B组均最高。结论 PRRS活疫苗与灭活疫苗组合在仔猪上的免疫效果优于PRRS疫苗组合组，B厂家疫苗免疫组免疫效果优于A厂家免疫组。
- 疫苗组合 /
- PRRS活疫苗 /
- PRRS灭活疫苗 /
- 免疫效果
Abstract: Objective Four combinations of Porcine reproductive and respiratory syndrome (PRRS) vaccine including live vaccine and inactivated vaccine were designed to improve the best immune effect for piglets, which would provide the reference for the formulation of PRRS vaccine prevention and control program. Methods A total of 25 piglets aged 28–35 days were randomly divided into 5 groups. In group A, piglets were firstly immunized with the live vaccine of PRRS (CH-1R strain) from manufacturer A and secondly immunized with the inactivated vaccine of PRRS (CH-1a strain ) from manufacturer A at an interval of 14 days. In group B, they was firstly immunized with the live vaccine of PRRS (strain CH-1R) B and secondly inactivated with the inactivated vaccine of PRRS (CH-1a strain) from manufacturer B, respectively. In group C, they was immunized for two times with the inactivated vaccine of PRRS (CH-1a strain) from manufacturer A. The interval of two immunization was 14 days, and 2 mL vaccine was injected into each piglet muscle. In group D, they was immunized for two times with the inactivated vaccine of PRRS (CH-1a strain) from manufacturer B. Group E was the control. The levels of cytokines (IL-2, IFN-γ, IFN-α, TNF-α) in peripheral blood were detected before and 1, 3, 5, 7 days after immunization. Blood samples were collected before the first immunization, 14 days after the first immunization, and 7, 14, 21 and 28 days after the second immunization to detect the levels of antibodies anti-GP5 protein and N protein. All groups were challenged with PRRSV at the 28^th day after the second immunization, and body temperature was detected for 21 days after PRRSV challenge, and necropsy was performed at the 21^st day after the PRRSV challenge. Results The test results showed that the mRNA expression levels of cytokines in all groups showed different change trend in control group after first immunization, the IL-2 expression levels decreased along with the increase of day age, but it in group A, B were significantly higher than those of group C, D, E at the 3^rd day; the expression level of IFN-γ in groups C and D were significantly higher than those in groups A and B at 3^rd day, 5^th day and 7^th day; the expression level of IFN-α in group B was significantly higher than that in other groups at the 3^rd day, 5^th day and 7^th day, and the expression level of TNF-α in group A was significantly higher than those in other groups at the 7^th day. After PRRSV wide strain challenge, the temperature of the control group rose quickly and high, and the duration of high temperature was long. The temperature of piglets in groups B increased less, and quickly returned to normal temperature. After immunization, the antibody level, antibody positive rate and challenge protection rate were the highest in group B. Conclusion The immune effect of live vaccine and inactivated vaccine of PRRS combination immunization is better than that of inactivated vaccine combination immunization, and the immune effect of vaccine from manufacturer B is better than that from manufacturer A.
- different vaccine combinations /
- live vaccine of PRRS /
- inactivated vaccine of PRRS /
- immune effect

HTML全文

图 1 不同时间点各组IL-2含量变化

（1）不同时间点各组比较用小写字母表示，不同字母代表差异显著（P＜0.05）；（2）A、B、C、D、E表示不同免疫组，其免疫信息见表1。图2、3、4同。

Figure 1. Changes of IL-2 content in each group at different time points

1）Comparisons between groups at different time points were represented by lowercase letters, with different letters representing significant differences (P＜0.05). 2) A, B, C, D and E represent different immune groups, and their immune information is shown in Table 1. The same as Fig. 2, 3, 4.

下载: 全尺寸图片幻灯片

图 2 不同时间点各组IFN-γ含量变化

Figure 2. Changes of IFN-γ content in each group at different time points

下载: 全尺寸图片幻灯片

图 3 不同时间点各组IFN-α含量变化

Figure 3. Changes of IFN-α content in each group at different time points

下载: 全尺寸图片幻灯片

图 4 不同时间点各组TNF-α含量变化

Figure 4. Changes of TNF-α content in each group at different time points

下载: 全尺寸图片幻灯片

图 5 不同组免后GP5蛋白抗体水平

1-14 d代表一免后14 d，2-7 d代表二免后7 d，2-14 d，代表二免后14 d，2-21 d，代表二免后21 d，2-28 d代表二免后28 d。

Figure 5. GP5 protein antibody levels in different groups after immunization

1-14 d represents the 14^th day after the first immunization，2-7 d represents the 7^th day after the second immunization，2-14 d represents the 14^th day after the second immunization，2-21 d represents the 21^st day after the second immunization，2-28 d represents the 28^th day after the second immunization.

下载: 全尺寸图片幻灯片

图 6 不同组免后N蛋白抗体水平

1-14 d代表1免后14 d，2-7 d代表2免后7 d，2-14 d，代表2免后14 d，2-21 d，代表2免后21 d，2-28 d代表2免后28 d

Figure 6. N protein antibody levels in different groups after immunization

下载: 全尺寸图片幻灯片

图 7 各组攻毒后体温变化曲线

Figure 7. Body temperature change curve of each group after challenge

下载: 全尺寸图片幻灯片

图 8 各组攻毒后肺脏病变情况

Figure 8. Lung lesions in each group after challenge

下载: 全尺寸图片幻灯片

表 1 各组免疫信息

Table 1. Information table of immune in each group

组别 Group	数量 Total /头	第一次免疫 The first immunization			第二次免疫（一免后14 d） Second immunization (14 days after the first immunization)
组别 Group	数量 Total /头	疫苗类型 Vaccines	免疫方法 Immune type	剂量 Dose/(mL·头⁻¹)	疫苗类型 Vaccines	免疫方法 Immune type	剂量 Dose/(mL·头⁻¹)
A组 A Group	5	PRRS活疫苗（A）	肌注	2	PRRS灭活疫苗（A）	肌注	2
B组 B Group	5	PRRS活疫苗（B）	肌注	2	PRRS灭活疫苗（B）	肌注	2
C组 C Group	5	PRRS灭活疫苗（A）	肌注	2	PRRS灭活疫苗（A）	肌注	2
D组 D Group	5	PRRS灭活疫苗（B）	肌注	2	PRRS灭活疫苗（B）	肌注	2
E组 E Group	5	灭菌生理盐水	肌注	2	灭菌生理盐水	肌注	2

下载: 导出CSV

表 2 相关引物序列表

Table 2. Sequence table of related primers

引物名称 Primers names	引物序列（5′-3′） Sequences (5′-3′)
GAPDH-F	TATGATTCCACCCACGGCAAG
GAPDH-R	CCACAACATACGTAGCACCAG
IL-2-F	AAGATGCAGCTCTTGTGTTGC
IL-2-R	TCAACAGCAGTTACTGTCTCA
IFN-α-F	AATCTCTCCCTTCTCCTGCCT
IFN-α-R	CAGGAGGAAGAATGGGCTTGT
IFN-γ-F	CAGAAGCTAACTCTCTCCGAA
IFN-γ-R	TCTGACTTCTCTTCCGCTTT
TNF-α-F	GAAGACACACCCCCGAACAGG
TNF-α-R	CGGCACTGAGTCGATCATCC

下载: 导出CSV

表 3 不同疫苗组免疫后GP5蛋白抗体阳性率

Table 3. Positive rates of antibodies against GP5 proteins after immunization in different groups （单位：%）

组别 Group	一免前 Before first immunization	一免后14 d 14^th d after first immunization	二免后7 d 7^th d after second immunization	二免后14 d 14^th d after second immunization	二免后21 d 21^th d after second immunization	二免后28 d 28^th d after second immunization
A组 Group A	0	0	20	60	80	100
B组 Group B	0	0	20	80	100	100
C组 Group C	0	0	0	0	20	20
D组 Group D	0	0	20	20	40	60
E组（对照）Group E (Control)	0	0	0	0	0	0

下载: 导出CSV

表 4 不同疫苗组免疫后N蛋白抗体阳性率

Table 4. Positive rate of N protein antibody in different groups after immunization （单位：%）

组别 Group	一免前 Before the first immunization	一免后14 d 14^th d after the first immunization	二免后7 d 7^thd after second immunization	二免后14 d 14^thd after second immunization	二免后21 d 21^thd after second immunization	二免后28 d 28^th d after second immunization
A组 Group A	0	0	40	60	100	100
B组 Group B	0	0	60	80	100	100
C组 Group C	0	0	0	20	20	40
D组 Group D	0	0	20	40	60	60
E组（对照）Group E (Control)	0	0	0	0	0	0

下载: 导出CSV

表 5 不同组攻毒保护率

Table 5. Challenge protection rate of different groups

组别 Group	攻毒保护率 Challenge protection rate/%
A组 Group A	80
B组 Group B	100
C组 Group C	40
D组 Group D	60
E组（对照） Group E（control）	0

下载: 导出CSV

参考文献(16)

[1]	殷震, 刘景华. 动物病毒学[M]. 2版. 北京: 科学出版社, 1997: 619-625.
[2]	郭宝清, 蔡雪晖, 刘文兴, 等. 猪繁殖与呼吸综合征灭活疫苗的研制 [J]. 中国预防兽医学报, 2000(4):259−262. GUO B Q, CAI X H, LIU W X, et al. Development of oil emulsion inactivated vaccine of Porcine reproductive and respiratory syndrome [J]. Chinese Journal of Preventive Veterinary Medicine, 2000(4): 259−262.(in Chinese)
[3]	李丽琴. 高致病性猪蓝耳病弱毒疫苗与灭活苗免疫特性的研究[D]. 北京: 中国农业科学院, 2010. LI L Q. Study on immune characteristics of attenuated vaccine and inactivated vaccine of highly pathogenic PRRS[D]. Beijing: Chinese Academy of Agricultural Sciences, 2010. (in Chinese)
[4]	梁皓仪. 不同蓝耳病疫苗的免疫效果—抗体效价分析 [J]. 养猪, 2008(4):70−72. LIANG H Y. Immune effect of different vaccines for blue ear disease-analysis of antibody titers [J]. Swine Production, 2008(4): 70−72.(in Chinese)
[5]	张书存, 王志鸿, 廖智慧. 高致病性猪蓝耳病的防控离不开弱毒疫苗免疫 [J]. 中国猪业, 2012(9):29−34. ZHANG S C, WANG Z H, LIAO Z H. The prevention and control of highly pathogenic porcine blue ear disease can not be separated from attenuated vaccine immunization [J]. China Swine Industry, 2012(9): 29−34.(in Chinese)
[6]	梁莹, 杨莉. 高致病性蓝耳病弱毒疫苗及灭活疫苗免疫效果的比较 [J]. 中国动物检疫, 2012(11):57−58. LIANG Y, YANG L. Comparison of immune effects between attenuated vaccine and inactivated vaccine against highly pathogenic blue ear disease [J]. China Animal Health Inspection, 2012(11): 57−58.(in Chinese)
[7]	刘义, 杨秀芬, 冯云飞, 等. 猪繁殖与呼吸综合征病毒N蛋白基因工程亚单位疫苗免疫保护试验 [J]. 黑龙江畜牧兽医, 2018(5):174−177,267,268. LIU Y, YANG X F, FENG Y F, et al. Immune protection test of porcine reproductive and respiratory syndrome virus N protein gene engineering subunit vaccine [J]. Heilongjiang Animal Science and Veterinary Medicine, 2018(5): 174−177,267,268.(in Chinese)
[8]	秦学远, 李金海, 罗毅, 等. 高致病性猪蓝耳病灭活疫苗和弱毒疫苗免疫效果试验 [J]. 黑龙江畜牧兽医(下半月), 2014(1):61−63. QIN X Y, LI J H, LUO Y, et al. Experimental study on immune effect of inactivated vaccine and attenuated vaccine against highly pathogenic PRRS [J]. Heilongjiang Animal Science and Veterinary Medicine, 2014(1): 61−63.(in Chinese)
[9]	刘海珍, 张俊文. 不同猪蓝耳病疫苗对仔猪免疫效果对比试验 [J]. 中国畜禽种业, 2013(2):42−43. LIU H Z, ZHANG J W. Comparative experiment on immune effect of different porcine blue ear disease vaccines on piglets [J]. The Chinese Livestock and Poultry Breeding, 2013(2): 42−43.(in Chinese)
[10]	杨竹鸣. pVAX1-IFN-α作为猪蓝耳病疫苗免疫佐剂的评价[D]. 南京: 南京农业大学, 2012. YANG Z M. Evaluation of PVX1-IFN-α as immune adjuvant of Porcine blue ear disease vaccine[D]. Nanjing: Nanjing Agricultural University, 2012. (in Chinese)
[11]	GONIN P, PIRZADEH B, GAGNON C A, et al. Se- roneutralization of porcine reproductive and respirato-ry syndrome virus correlates with antibody response to the GP5 major envelope glycoprotein [J]. J Vet Di- agn Invest, 1999, 11(1): 20−26. doi: 10.1177/104063879901100103
[12]	冯延, 郭嘉, 许瑞勤, 等. 针对不同形式猪繁殖与呼吸综合征病毒GP5重组蛋白的抗体中和活性比较 [J]. 畜牧兽医学报, 2018(6):1222−1230. FENG Y, GUO J, XU R Q, et al. Comparison of neutralizing antibodies induced by PRRSV GP5 recombinant proteins in different forms of truncation [J]. Chinese Journal of Animal and Veterinary Sciences, 2018(6): 1222−1230.(in Chinese)
[13]	邢刚. 猪蓝耳病病毒的结构与相关抗体的分析 [J]. 中国牧业通讯, 2011(12):74. XING G. Analysis of the structure and related antibodies of PRRSV [J]. China Animal Husbandry Bulletin, 2011(12): 74.(in Chinese)
[14]	任向阳, 吴文福, 岑小清, 等. 猪瘟活疫苗(传代细胞源)免疫产生期和免疫持续期试验 [J]. 中国兽医杂志, 2017(4):92−94,98. REN X Y, WU W F, CEN X Q, et al. The onset time and maintenance phase of immunity protection for classical swine fever live vaccine(cell line origin) [J]. Chinese Journal of Veterinary Medicine, 2017(4): 92−94,98.(in Chinese)
[15]	WILLIAM L,MENGELING. The porcinereproductive and respiratory syndromequandary Part one Fact versus speculation [J]. Journal of Swine Health and Production, 2005, 13(2): 91−95.
[16]	严玉霖, 高洪. 猪繁殖与呼吸综合征诊断技术研究进展 [J]. 动物医学进展, 2008(7):81−85. YAN Y L, GAO H. Advance in Diagnostic Techniques for Porcine reproductive and respiratory syndrome virus [J]. Progress in Veterinary Medicine, 2008(7): 81−85.(in Chinese)