Histamine Release Factor from Dermanyssus gallinae (De Geer): Characterization and in vitro assessment as a protective antigen
Histamine Release Factor from Dermanyssus gallinae (De Geer): Characterization and in vitro assessment as a protective antigen
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DOI：
10.1016/j.ijpara.2008.09.006
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发表时间：
2009-03-01
影响因子：
4
通讯作者：
Huntley, John F.
中科院分区：
文献类型：
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作者：
Bartley, Kathryn;Nisbet, Alasdair J.;Huntley, John F.
从食血家禽革螨（Dermanyssus gallinae）中鉴定出一段编码蜱组胺释放因子（HRF）的同源基因，全长174个氨基酸。预测的鸡D. HRF蛋白（Dg-HRF-1）序列与蜱虫HRF高度保守（一致性为52-54%），与来自哺乳动物和其他无脊椎动物的翻译控制肿瘤蛋白（TCTP）在较小程度上保守（范围为38-47%）。在系统发育上，Dg-HRF-1与蜱的HRF进化支划分，这表明它们具有共同的谱系和潜在的相似功能。制备了重组Dg-HRF-1蛋白（rDg-HRF-1），并在体外诱导大鼠腹膜肥大细胞脱颗粒，证实鸡鸡鸡保留了组胺释放功能。在家兔和蛋鸡中制备了抗rDg-HRF-1的多克隆抗体。免疫印迹法证实天然Dg-HRF是一种可溶性蛋白，螨体切片免疫组化染色显示Dg-HRF虽然普遍存在，但多见于螨的生殖、消化和神经节组织。一项对连续饲养在螨虫感染的商业家禽单位的母鸡的调查未能发现重组或天然Dg-HRF的IgY特异性，这表明Dg-HRF在感染/饲养期间不暴露于宿主，因此可能有潜力使用隐藏抗原方法作为疫苗。为了测试rDg-HRF-1的保护能力，我们在新鲜肝素化的鸡血液中富集蛋黄来源的抗rdg - hrf IgY抗体，并用体外饲养系统喂半饥饿的螨虫。用抗dg - hrf IgY饲喂一餐血后，螨虫死亡率有统计学意义的增加（P = 0.004）。据我们所知，这项工作首次证明了用重组鸡D.鸡抗原接种母鸡控制螨感染的可行性，以及rDg-HRF-1作为疫苗抗原的潜力。(C) 2008澳大利亚寄生虫学学会Elsevier Ltd.出版。版权所有。
A cDNA encoding a 174-amino-acid orthologue of a tick histamine release factor (HRF) was identified from the haematophagous poultry red mite Dermanyssus gallinae. The predicted D. gallinae HRF protein (Dg-HRF-1) sequence is highly conserved with the tick HRFs (identity 52-54%) and to a lesser degree with translationally controlled tumour proteins (TCTP) from mammals and other invertebrates (range 38-47%). Phylogenetically, Dg-HRF-1 partitions with the tick HRF clade Suggesting a shared linage and potentially similar function(s). A recombinant Dg-HRF-1 protein (rDg-HRF-1) was produced and shown to induce degranulation of rat peritoneal mast cells in vitro, confirming conservation of the histamine-releasing function in D. gallinae. Polyclonal antibodies were generated in rabbits and hens to rDg-HRF-1. Western blotting demonstrated that native Dg-HRF is a soluble protein and immunohistochemical staining of mite sections revealed that the distribution of Dg-HRF, although ubiquitous, is more common in mite reproductive, digestive and synganglion tissues. A survey of hens housed continuously in a mite-infested commercial poultry unit failed to identify IgY specific for recombinant or native Dg-HRF, indicating that Dg-HRF is not exposed to the host during infestation/feeding and may therefore have potential as a vaccine using the concealed antigen approach. To test the protective capability of rDg-HRF-1, fresh heparinised chicken blood was enriched with yolk-derived anti-Dg-HRF IgY antibodies and fed to semi-starved mites using an in vitro feeding system. A statistically significant increase in mortality was shown (P = 0.004) in mites fed with anti-Dg-HRF IgY after just one blood meal. The work presented here demonstrates, to our knowledge for the first time, the feasibility of vaccinating hens with recombinant D. gallinae antigens to control mite infestation and the potential of rDg-HRF-1 as a vaccine antigen. (C) 2008 Australian Society for Parasitology Inc. Published by Elsevier Ltd. All rights reserved.