猪繁殖与呼吸综合征弱毒活疫苗免疫猪脾脏转移因子的制备及其活性分析  

Preparation and Immunological Activities of Transfer Factor from the Spleen of Pig Immunized with Porcine Reproductive and Respiratory Syndrome Attenuated-live Virus Vaccine

作　　者：郭海勇[1] 袁洪兴[2] 孙泰然 曹静 宋勤叶[2] 李潭清[2] 逯纪成 GUO Hai-yong;YUAN Hong-xing;SUN Tai-ran;CAO Jing;SONG Qin-ye;LI Tan-qing;LU Ji-cheng(School of Life Science,Jilin Normal University,Siping 136000,China;Hebei Province Research Center of Veterinary Biologicals Engineering and Technology,College of Veterinary Medicine,Hebei Agricultural University,Baoding 071001,China;The Center for Animal Disease Control and Prevention,Baoding 071000,China)

机构地区：[1]吉林师范大学生命科学学院,四平136000 [2]河北农业大学动物医学院河北省兽用生物制品工程技术研究中心,保定071001 [3]河北省保定市动物疫病预防控制中心,保定071000

出　　处：《畜牧兽医学报》2018年第7期1467-1474,共8页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：河北省科技计划项目(17226613D-2;13226603D-1〈2013-X〉); 河北省生猪产业技术体系疫病控制专项

摘　　要：从猪繁殖与呼吸综合征(PRRS)弱毒活疫苗免疫猪脾制备转移因子(TF),检测其免疫活性,为开发新型免疫增强剂和有效防制PRRS提供科学依据。用PRRSV弱毒活疫苗免疫健康育成猪,无菌采集脾,通过透析法从脾提取TF。应用磺基水杨酸法、茚三酮反应和硝酸-钼酸铵试验等方法检测TF的主要成分,经高效液相色谱法测定TF中氨基酸种类及其含量;通过鸡红细胞吞噬试验和MTT法,检测TF对小鼠腹腔巨噬细胞吞噬功能和外周血淋巴细胞增殖活性的调节作用。进而将9头4周龄健康仔猪随机分为A、B和C组,每组3头。A组每头猪左、右侧耳后分别肌内注射1头份PRRSV疫苗和1mL TF;B组每头猪注射1头份PRRSV疫苗与1mL TF的混合液;C组每头猪注射1头份PRRSV疫苗与1mL生理盐水混合液。免疫后20d,通过MTT法检测每组猪外周血淋巴细胞(PBLC)的增殖活性。制备的TF内无蛋白质成分,含有核酸和18种氨基酸,每毫升TF含核酸21.5μg、总氨基酸3.479mg。TF能显著增强小鼠腹腔巨噬细胞对鸡红细胞的吞噬能力和PBLC增殖活性。当TF与PRRSV疫苗分开或混合注射给4周龄仔猪后,能够显著提高仔猪PBLC的增殖能力。从PRRS弱毒活疫苗免疫猪的脾制备了TF,其能显著提高巨噬细胞的吞噬功能和外周血淋巴细胞的增殖活性。The objective of this study was to prepare transfer factor（TF）from the spleen of pig immunized with porcine reproductive and respiratory syndrome（PRRS）attenuated-live virus vaccine,then detect its immune activities and expect to provide a scientific basis for developinganew immunopotentiator and prevention and control of PRRS.TF was extracted from spleen collected from healthy grown pigs which had been vaccinated with PRRS attenuated-live virus vaccine.The main components of TF were detected using sulfonyl salicylic acid,ninhydrin and ammonium molybdate-ammonium nitrate reactions,and kinds and contents of amino acids were measured by high performance liquid chromatography.The regulating effects on phagocytosis of peritoneal macrophage and lymphocytic proliferation of mice were checked by chicken erythrocyte phagocytosis test and MTT assay,respectively.Furthermore,nine piglets were divided into group A,B and C of 3 each.The piglets in the group A were injected intramuscularly with one dose of PRRS attenuated-live virus vaccine in left side opisthotic part and 1 mL of TF in the right side,respectively.Each one in the group B was injected intramuscularly the mixed liquor of one dose of PRRS attenuated-live virus vaccine with 1 mL of TF,and in the group C was did the mixed liquor of one dose of PRRS attenuated-live virus vaccine with 1 mL of normal saline.The proliferative activity of porcine peripheral blood lymphocyte（PBLC）from each group was measured by MTT on day 20 post-inoculation.There were nucleic acid and 18 kinds of amino acids in TF prepared in this study,containing 21.5μg of nucleic acid and 3.479 mg total amino acids per mL of TF.Phagocytic ability of peritoneal macrophage to chicken erythrocytes and proliferative activity of PBLC from mice were enhanced significantly following stimulation of TF.Moreover,the proliferation capacity of porcine PBLC was also increased remarkably when the four-week-old piglets were injected with PRRS attenuated-live virus vaccine and TF simultaneously in differe

关 键 词：猪繁殖与呼吸综合征病毒 猪 脾 转移因子 免疫活性 

分 类 号：S852.659.6[农业科学—基础兽医学]