旨在预防猫、犬诱导的过敏反应,本研究将猫主要变应原Fel d 1与犬主要变应原Canf 1融合原核表达,纯化制备了 rFel d 1-Can f 1 融合变应原.将层状双氢氧化物(layered double hydroxides,LDH)与 rFel d 1-Can f 1混合,制备LDH-rFel d 1-Can f 1,皮下注射法免疫BALB/c小鼠,随后利用rFel d 1-Can f 1融合变应原诱导过敏小鼠模型,通过检测小鼠应激性过敏反应的体温变化、耳朵点刺试验、肺部组织切片HE染色、血清IgE水平等指标评价LDH-rFel d 1-Can f 1对猫犬诱发过敏反应的预防效果.研究表明,LDH与rFel d 1-Can f 1质量比为7∶1时,LDH可以完全吸附rFel d 1-Can f 1融合蛋白.LDH-rFel d 1-Can f 1可以有效缓解应激性过敏反应引起的小鼠体温下降,显著减少过敏导致的染料渗透面积.肺部组织切片HE染色结果显示,LDH-rFel d 1-Can f 1可以缓解过敏引起的炎症细胞浸润.此外,LDH-rFel d 1-Can f 1免疫后可以诱导小鼠产生较高水平IgG,这为开发抑制宠物猫犬诱发过敏反应的疫苗奠定基础.
Preventive Effect of Layered Double Hydroxide-rFel d 1-Can f 1 Fusion Allergens on Anaphylaxis in Mouse Model
To prevent allergic reactions induced by cats and dogs,we purified rFel d 1-Can f 1 fusion allergen by fusion prokaryotic expression of cat primary allergen Fel d 1 and dog primary allergen Can f 1.We mixed Layered double hydroxides(LDH)with rFel d 1-Can f 1 to prepare LDH-rFel d 1-Can f 1 and immunized BALB/c mice by subcutaneous injection.Subsequently,we used rFel d 1-can f 1 fusion allergen to induce allergic mice models.We evaluated the preventive effect of LDH-rFel d 1-Can f 1 on allergic reactions caused by cats and dogs through anaphylaxis,ear prick test,H&E staining of lung tissue sections,and serum IgE level.The results showed that LDH could completely adsorb the fusion protein of rFel d 1-Can f 1 when the mass ratio of LDH to rFel d 1-Can f 1 was 7∶1.The LDH-rFel d 1-Can f 1 can effectively relieve the hypothermia caused by stress allergic reaction in mice,and significantly reduce the dye penetration area caused by allergy.H&E staining of lung tissue sections showed that LDH-rFel d 1-Can f 1 could relieve inflammatory cell invasion caused by the allergy.In addition,the LDH-rFel d 1-Can f 1 can induce mice to produce higher levels of IgG,which lays the foundation for the development of vaccines that inhibit allergic reactions induced by pet cats and dogs.
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