Antibody Response of Dogs After Immunisation with Chimeric Vaccine Against Borreliosis

J. Nepereny , V. Vrzal , M. Raska , E. Weigl , M. Krupka , J. Turanek , J. Masek
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引用次数: 2

Abstract

Two chimeric recombinant fusion proteins (ch-rOspC and ch-rOspA) were created. They are composed of the immunodominant domains of OspC and OspA proteins described in the clinically most important strains of Borrelia. The gene constructs for these chimeric proteins were inserted into plasmids pET28 allowing induced gene expressions in a bacterial system.

The proteins were expressed in E. coli BL21 strains, purified and used for preparation of the vaccine.

One dose of the tested vaccines contained 50 μg of each relevant protein (ch-rOspC, ch-rOspA, or ch-rOspC + ch-rOspA). PET GEL A (Seppic) or Aluminium hydroxide gel as the immune adjuvants were used.

The dogs were vaccinated three times at 21 days intervals subcutaneously or intradermally and unvaccinated controls were also included.

The vaccine-elicited serum action antibodies specific to OspA and OspC were determined using in-house ELISA sets.

The immunisation induced specific antibody response in the vaccinated animals and OspC and OspA from representative genospecies B. garinii, B. afzelii, and B. burgdorferi sensu stricto were recognized. The control dogs were without antibody response.

ELISA examination enables determination of specific post-vaccination antibodies against OspA and OspC. Detection of these antibodies and their quantification may be used for evaluation of efficiency of vaccines.

犬用嵌合疫苗免疫疏螺旋体病后的抗体反应
构建了两个嵌合重组融合蛋白ch-rOspC和ch-rOspA。它们由临床最重要的伯氏疏螺旋体菌株中描述的OspC和OspA蛋白的免疫优势结构域组成。这些嵌合蛋白的基因结构被插入到质粒pET28中,允许在细菌系统中诱导基因表达。这些蛋白在大肠杆菌BL21株中表达,纯化后用于制备疫苗。每种相关蛋白(ch-rOspC、ch-rOspA或ch-rOspC + ch-rOspA)各含50 μg。采用PET GEL A (Seppic)或氢氧化铝凝胶作为免疫佐剂。这些狗每隔21天注射三次皮下或皮内疫苗,未接种疫苗的对照组也包括在内。疫苗诱导的OspA和OspC特异性血清作用抗体采用内部ELISA试剂盒测定。免疫接种动物和典型基因种B. garinii、B. afzelii和B. burgdorferi的OspC和OspA均产生特异性抗体反应。对照犬无抗体反应。ELISA检测能够确定接种后针对OspA和OspC的特异性抗体。这些抗体的检测和定量可用于评估疫苗的效率。
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