【作者】 徐磊；

【导师】 吴宝成； 黄瑜； 张渊魁；

【作者基本信息】 福建农林大学 ， 临床兽医学， 2009， 硕士

【摘要】 猪瘟(Classical Swine Fever,CSF)是由猪瘟病毒引起猪的一种高度接触性传染病,给世界养猪业造成了巨大经济损失。在我国,猪瘟依然是危害养猪业最为严重的疫病之一。目前在我国养猪业中,多应用兔化弱毒脾淋苗或细胞苗预防猪瘟。然而,在猪瘟抗体水平较高的猪群中,有的仍发生猪瘟。为评价这两种疫苗的细胞免疫效果,进行了比较研究。本试验采用常规白细胞计数、流式细胞技术、淋巴细胞转化试验、ELISA试剂盒对猪瘟兔化弱毒细胞苗免疫仔猪(以下简称细胞苗组)、脾淋苗免疫仔猪(有两组,以下分别简称脾淋苗A组、脾淋苗B组)和注射生理盐水的仔猪(简称对照组)分别进行主要免疫相关指标的测定,结果如下:1、猪外周血CD3+、CD4+、CD8+亚群分型测定免疫后第1、7、14、24和35天细胞苗组CD3+淋巴细胞比例低于两个脾淋苗组,但差异不显著;CD4+淋巴细胞亚群比例由高至低依次为:两个脾淋苗组、对照组、细胞苗组,其中于第1天差异显著;组内各时间点CD4+/CD8+比值只有细胞苗组差异显著,且细胞苗组CD4+/CD8+比值始终低于两个脾淋苗组,其中于第35天差异显著。2、猪外周血淋巴细胞增殖活性测定免疫后第1、3、7、14、24和35天植物血凝素非特异性淋巴细胞活力由高至低依次为:脾淋苗A组、脾淋苗B组、细胞苗组和对照组,其中于第3和7天差异显著;免疫后7天起猪瘟兔化弱毒特异性淋巴细胞活力由高至低依次为:脾淋苗A组、脾淋苗B组、细胞苗组和对照组,其中于第14天差异显著。3、猪外周血细胞因子水平测定免疫后第1、3、7、14、24和35天IFN-γ水平由高到低依次为:脾淋苗A组、脾淋苗B组、细胞苗组和对照组,其中于第1、3、7、14和24天差异显著;脾淋苗A组IL-3水平始终高于其他组,而对照组始终最低。细胞苗组IL-3水平于免疫后7天内低于脾淋苗B组,7天后高于脾淋苗B组。其中于第1、3、7、14、24和35天差异显著。4、血常规检测猪外周血白细胞、淋巴细胞总数组间差异不显著,且组内各日龄差异亦不显著。综合以上试验结果可见,与猪瘟兔化弱毒细胞苗相比,猪瘟兔化弱毒脾淋苗免疫猪具有高比例的CD3+和CD4+淋巴细胞亚群,高而稳定的CD4+/CD8+比值,高活力的淋巴细胞以及高水平的IFN-γ和IL-3,表明猪瘟兔化弱毒脾淋苗的细胞免疫效果更为明显,为猪瘟疫苗的选择提供了科学依据。更多还原

【Abstract】 Classical swine fever (CSF) is a highly contagious disease in pigs caused by the CSF virus. It brings a severe economic losses to the worldwide pig farming. In China it is still one of the most severe diseases. Now Both hog cholera lapinized virus (HCLV) rabbit spleen and lymph tissue origin vaccine and HCLV bovine testis cell vaccine were spreadly used to prevent pigs from CSF in pig farming of our country’s. But pigs containing high antibody against CSFV still suffer from CSF. In order to estimate cell immunity efficiency of CSFV vaccine, we have done some research on comparing them .In this study, we develop a track observation for some principal references of immunity in order to take a objective compared analysis with HCLV bovine testis cell vaccine group, HCLV rabbit spleen and lymph tissue origin vaccine group A , HCLV rabbit spleen and lymph tissue origin vaccine group B and negative group by the traditional methods of white blood counting, Flow cytometry,lymphocytes proliferation test and ELISA test kit.The results were shown as follows:1 the level of CD3+,CD4+,CD8+ lymphocytes subsets in PBMC On 1,7,14,24 and 35 days after immunization CD3+/lym of HCLV bovine testis cell vaccine group is lower than HCLV rabbit spleen and lymph tissue origin vaccine group A and B ,but there is no significant difference; CD4+/lym from high to low respectively is: HCLV rabbit spleen and lymph tissue origin vaccine group A and B,negative group and HCLV bovine testis cell vaccine group .And there is a significant difference on 1 day after immunization; There is a significant difference on the ratio of CD4+/ CD8+ only for HCLV bovine testis cell vaccine group between different days.And its ratio of CD4+/ CD8+ is lower than HCLV rabbit spleen and lymph tissue origin vaccine group A and B.There is a significant difference on 35 days after immunization.2 Result of lymphocytes proliferation test On 1,3,7,14,24 and 35 days after immunization SI of nonspecific proliferation from high to low respectively is: group A, group B, HCLV bovine testis cell vaccine group and negative group , and there is a significant difference on 3 and 7 days after immunization; After immunization for 7 days SI of specific proliferation from high to low respectively is: group A, group B, HCLV bovine testis cell vaccine group and negative group,and there is a significant difference on 14 days after immunization.3 The level of some cytokine On 1,3,7,14,24 and 35 days after immunization the level of IFN-γfrom high to low respectively is: group A, group B, HCLV bovine testis cell vaccine group and negative group ,and there is a significant difference on 1,3,7,14 and 24 days after immunization;The level of IL-3 from pigs of group A is higher than other group’s, meanwhile the level of IL-3 from pigs of negative group is lower than other group’s. The level of IL-3 from pigs of HCLV bovine testis cell vaccine group is lower than group B , but after immunization for 7 days it is higher than group B. And there is a significant difference on 1,3,7,14,24 and 35 days after immunization.4 Result of white blood counting there is no significant difference between groups and different days on leucocyte and lymphocyte count .The results demonstrat that compaired with HCLV bovine testis cell vaccine, the pigs vaccinated with HCLV rabbit spleen and lymph tissue origin vaccine contain high CD3+/lym and CD4+/lym, high and stable ratio of CD4+/ CD8+ ,high activity of T lymphocytes and high level of IFN-γand IL-3.So the cell immunity efficiency of HCLV rabbit spleen and lymph tissue origin vaccine is much more obviously .The objective evaluation take a scientific dependence for choosing CSFV vaccine .更多还原