摘要14

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	不同剂量小鹅瘟VP3基因疫苗在雏鹅体内的表达时相和分布规律

	黎敏¹，朱德康^1，2，卢菲¹，韩新锋¹，程安春^1,2，汪铭书^1，2，刘晓东¹，车茜¹，陈孝跃^1，2

	（1.四川农业大学动物医学院禽病防治研究中心,四川雅安625014； 2.动物疫病与人类健康四川省重点实验室，四川雅安625014)

	摘要：
	将小鹅瘟基因疫苗（pcDNA-GPV-VP3）分别以每只200、100和50μg肌肉注射免疫30日龄四川白鹅，以生理盐水和空载体质粒pcDNA3.1(+)为对照，于免疫接种后第12h、1d、3d、7d、21d、35d、63d和105d采集各组织器官，用免疫组织化学方法检测pcDNA-GPV-VP3在雏鹅各组织器官中的表达和分布。结果显示，各剂量免疫组第1d在免疫部位肌肉，第3d在心肌中均能检测到pcDNA-GPV-VP3的表达产物，第7d的表达量最大，表达可持续至第63d；第3d时各剂量免疫组均能在各肠段的肠腺、肠绒毛的杯状细胞和肌细胞中检测到pcDNA-GPV-VP3的表达产物，第7d的表达量最多，表达可持续至第35d；肝、肺、肾、脾、胰腺、脑、腔上囊、胸腺中未见表达；不同剂量的pcDNA-GPV-VP3免疫雏鹅后各组织器官中表达产物的量和持续时间的总体规律为200μg＞100μg＞50μg，但非等比例递增。研究证实，pcDNA-GPV-VP3免疫雏鹅后能在免疫部位肌肉、心肌、小肠肠腺和肠绒毛的杯状细胞、肌细胞内表达。
	关键词：
	小鹅瘟VP3基因疫苗；肌肉注射；雏鹅；基因表达；免疫组织化学
	中图分类号：S 852.659.2 文献标识码： A 文章编号：1673-4696(2008)06-0514-07

Distribution and expression kinetics of different doses of GPV-VP3 DNA vaccine in gosling

LI Min¹，ZHU De-kang^1，2，LU Fei¹，HAN Xin-feng¹， CHENG An-chun^1，2,WANG Ming-shu^1，2, LIU Xiao-dong¹,CHE Qian¹,CHEN Xiao-yue^1，2

(1.Avian Diseases Research Center,College of Veterinary Medicine,Sichuan Agricultural University,Ya’an 625014,China;2.Key Laboratory of Animal Diseases and Human Health of Sichuan Province,Ya’an 625014,China)

Abstract:Forty 30-day-old Sichuan White geese in each experimental group were injected intramuscularly with gene vaccine pcDNA-GPV-VP3 against goose parvovirus VP3 at 200μg,100μg and 50μg per gosling,respectively.15 geese injected respectively with normal saline solution and plasmid backbone were set as controls.Different tissue samples were collected,respectively,at hour 12,and on day 1,3,7,21,35,63 and 105 post-immunization,and analyzed by indirect immunohistochemistry（ISH） for the expression and distribution of the pcDNA-GPV-VP3.Expression of the plasmid could be detected at the muscle injection sites in different doses groups on day 1 post-immunization and at the cardiac muscle on day 3 post-immunization,and the expression peaked on day 7 post-immunization and persisted till on day 63 post-immunization.Expression could be detected in the intestinal gland cells,goblet cells of the intestinal villus and the muscle cells,too,and the expression peaked on day 7 post-immunization and persisted on day 35 post-immunization.No expression was detected in liver,lung,kidney,spleen,pancreas,brain,bursa of Fabricius and thymus.The expression of the pcDNA-GPV-VP3 in the intramuscular injection group at 200μg per gosling was superior to those at 100μg or 50μg per gosling.The results demonstrated that pcDNA-GPV-VP3 could be expressed in the immunization sites,cardiac muscle,small intestinal gland cells,goblet cells of the intestinal villus and the muscle cells.

Key words: GPV-VP3 DNA vaccine；intramuscular injection；gosling；gene expression；immunohistochemistry


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