PRRSV VR2332弱毒株经滴鼻、注射途径接种仔猪的病毒血症和抗体产生的差异分析

doi:10.11843/j.issn.0366-6964.2019.02.015

摘要/Abstract

摘要：

为了解猪繁殖与呼吸综合征病毒（PRRSV）VR2332弱毒株经滴鼻和注射两种途径接种仔猪的病毒血症、抗PRRSV抗体以及抗PRRSV中和抗体的差异，选取20日龄健康仔猪9头，随机分3组，即滴鼻组、注射组和对照组。在感染后第0、3、7、11、15、19、23、27、31、35、39、43天前腔静脉采血，于第43天采血后摘取猪的脾、颌下淋巴结、腹股沟淋巴结和肠系膜淋巴结，并无菌收集猪肺泡巨噬细胞（PAM），通过实时荧光定量PCR方法检测病毒载量。用ELISA方法检测抗PRRSV抗体效价，用基于PAM的中和试验检测抗PRRSV中和抗体效价。结果显示：仔猪感染PRRSV后，①滴鼻组第3天只在2头猪血清中检测到PRRSV，第15-19天病毒血症达到高峰，病毒拷贝数均大于10⁴拷贝·μL^-1，第27天后病毒血症消失；注射组第3天从3头仔猪血清中均检测到病毒，在第15-23天病毒血症达到高峰，其中第19天病毒拷贝数大于10⁵拷贝·μL^-1，病毒血症从第31天后开始消失。②从几种淋巴结和PAM中均检测到PRRSV，且注射组的PRRSV拷贝数稍高于滴鼻组。③均在第27天从血清中检测到抗PRRSV抗体，随后抗体水平一直处于逐渐上升趋势，且第35-43天，注射组的抗PRRSV抗体水平高于滴鼻组。④滴鼻组在第35天之前抗PRRSV中和抗体效价都小于1：2，第43天抗PRRSV中和抗体效价为1：2；注射组在第3、11和19天的抗PRRSV中和抗体效价均小于1：2，第27、35、43天的中和抗体效价分别为1：4、1：8、1：4。试验结果表明，注射组的病毒拷贝数一直为滴鼻组的10~100倍，且病毒血症持续时间更长；注射组血清中抗PRRSV抗体产生的时间比滴鼻组早，并且抗体水平比滴鼻组高；滴鼻组抗PRRSV中和抗体水平很低，注射组产生的抗PRRSV中和抗体比滴鼻组早且抗体水平高。综上所述，VR2332弱毒株经注射免疫比滴鼻免疫能刺激猪产生更高水平的中和抗体，但同时也会引发更高水平的病毒血症。

Abstract:

The study was conducted to understand the difference of viremia, anti-PRRSV antibody and anti-PRRSV neutralizing antibody in piglets vaccinated against porcine reproductive and respiratory syndrome virus (PRRSV) VR2332 attenuated strain which was given by nasal drip and injection. Nine 20-day-old health piglets were randomly selected and separated into 3 groups which designated as nose dropping group, injection group and control group respectively. Blood samples were collected by the anterior vena cava at day 0, 3, 7, 11, 15, 19, 23, 27, 31, 35, 39, 43 after infection, spleen, submandibular lymph nodes, inguinal lymph nodes and mesenteric lymph nodes were obtained at day 43, PAMs were collected in a sterile room meanwhile. Viral load in piglets was detected with the real-time fluorescent quantitative PCR. The anti-PRRSV antibody titer was tested by ELISA, and the anti-PRRSV neutralizing antibody titer was detected by the method based on PAMs. The results showed that, ①PRRSV started to replicate in only 2 piglet's sera at day 3 after infection, viremia peaked at day 15-19, the virus copy number was greater than 10⁴ copies·μL^-1 and viremia disappeared after day 27 in nose dropping group. PRRSV started to replicate in all 3 piglet's serum at day 3 after infection, viremia peaked at day 15-23, the virus copy number was up to 10⁵ copies·μL^-1 at day 19 and viremia disappeared after day 31 in injection group. ②PRRRSV can be detected in these lymph nodes and PAMs, and the virus copy number of injection group was greater than nose dropping group. ③Anti-PRRSV antibody was produced in piglets at day 27 after infection from serum, and anti-PRRSV antibody levels of injection group was always higher than nose dropping group. ④Anti-PRRSV neutralizing antibody titers was 1:2 at day 43 and lower than 1:2 before day 35 in nose dropping group. Anti-PRRSV neutralizing antibody titers was 1:4 at day 27, 1:8 at day 35, 1:4 at day 43 and lower than 1:2 at day 3, 11 and 19 in injection group. The results showed that the virus copy number in injection group was always ten to one hundred times higher than that of nose dropping group, and duration of viremia was longer. Anti-PRRSV antibody producing time in injection group was earlier than nose dropping group, and antibody levels was higher. Anti-PRRSV neutralizing antibody levels in nose dropping group was very low, anti-PRRSV neutralizing antibody producing time in injection group was earlier than nose dropping group, and antibody levels were higher. In conclusion, higher level neutralizing antibody can be stimulated in piglets after inoculating PRRSV VR2332 attenuated strain by injection than by nasal drip, but higher level viremia also can be induced meanwhile.

中图分类号:

S858.285.3

李闻, 马思续, 李想通, 孙杨杨, 魏凤灵, 许瑞勤, 杨国宇, 夏平安, 张改平. PRRSV VR2332弱毒株经滴鼻、注射途径接种仔猪的病毒血症和抗体产生的差异分析[J]. 畜牧兽医学报, 2019, 50(2): 373-381.

LI Wen, MA Sixu, LI Xiangtong, SUN Yangyang, WEI Fengling, XU Ruiqin, YANG Guoyu, XIA Ping'an, ZHANG Gaiping. The Variance Analysis of Viremia and Antibody in Piglets Vaccinated against PRRSV VR2332 Attenuated Strain via Nasal Drip and Injection[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2019, 50(2): 373-381.

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