Identification and evaluation of vaccine candidate antigens from the poultry red mite (Dermanyssus gallinae).
Identification and evaluation of vaccine candidate antigens from the poultry red mite (Dermanyssus gallinae).
复制标题
DOI：
10.1016/j.ijpara.2015.07.004
复制
发表时间：
2015-11
影响因子：
4
通讯作者：
Nisbet AJ
中科院分区：
文献类型：
--
作者：
Bartley K;Wright HW;Huntley JF;Manson ED;Inglis NF;McLean K;Nath M;Bartley Y;Nisbet AJ
鉴定了10种家禽红螨疫苗候选抗原，并生产了重组版本。在饲喂来自接种疫苗的母鸡的血液后监测螨死亡率。观察到四种候选疫苗的螨死亡风险增加了1.6倍（P < 0.001）。最佳候选物包括：丝氨酸蛋白酶抑制剂、卵黄蛋白原、血红素糖蛋白和一种新蛋白。采用阴离子交换色谱法对食血性家禽体外寄生虫（鸡皮刺螨）的水提取物进行了亚分级。这些亚组分中的六个用于免疫母鸡，并将来自这些母鸡的血液在体外饲喂家禽红螨。这些饲料后的螨死亡率表明两个亚组分中存在保护性抗原，螨死亡的风险分别是对照组的3.1和3.7倍（P < 0.001）。二维免疫印迹和免疫亲和层析的组合，使用IgY从母鸡免疫这些亚组分，与蛋白质组学分析，以确定最强的免疫原性蛋白在每个这些亚组分。使用以下标准选择10种免疫反应性蛋白质作为候选疫苗进行评估：免疫识别强度;抗原暴露于血餐中抗体的可能性;邻苯二甲酸分子的拟定功能和已知疫苗潜力。这10种蛋白质中的每一种的重组版本在大肠杆菌中产生，并用于免疫母鸡。随后在体外用这些鸡的血液喂养螨表明，用Deg-SRP-1（丝氨酸蛋白酶抑制剂）、Deg-VIT-1（卵黄蛋白原）、Deg-HGP-1（血红素糖蛋白）或Deg-PUF-1（功能未知的蛋白质）免疫导致螨死亡风险显著增加（比用仅用佐剂免疫的对照母鸡的血液喂养的螨高1.7-2.8倍，P < 0.001）。在重组疫苗中使用这些抗原的潜力进行了讨论。
Ten poultry red mite vaccine candidate antigens were identified and recombinant versions produced. Mite mortality was monitored after feeding on the blood from vaccinated hens. A ⩾1.6-fold increased risk of mite death was observed with four of the vaccine candidates (P < 0.001). Best candidates include: a serpin, vitellogenin, hemelipoglycoprotein and a novel protein. An aqueous extract of the haematophagous poultry ectoparasite, Dermanyssus gallinae, was subfractionated using anion exchange chromatography. Six of these subfractions were used to immunise hens and the blood from these hens was fed, in vitro, to poultry red mites. Mite mortality following these feeds was indicative of protective antigens in two of the subfractions, with the risks of mites dying being 3.1 and 3.7 times higher than in the control group (P < 0.001). A combination of two-dimensional immunoblotting and immunoaffinity chromatography, using IgY from hens immunised with these subfractions, was used in concert with proteomic analyses to identify the strongest immunogenic proteins in each of these subfractions. Ten of the immunoreactive proteins were selected for assessment as vaccine candidates using the following criteria: intensity of immune recognition; likelihood of exposure of the antigen to the antibodies in a blood meal; proposed function and known vaccine potential of orthologous molecules. Recombinant versions of each of these 10 proteins were produced in Escherichia coli and were used to immunise hens. Subsequent in vitro feeding of mites on blood from these birds indicated that immunisation with Deg-SRP-1 (serpin), Deg-VIT-1 (vitellogenin), Deg-HGP-1 (hemelipoglycoprotein) or Deg-PUF-1 (a protein of unknown function) resulted in significantly increased risk of mite death (1.7–2.8 times higher than in mites fed blood from control hens immunised with adjuvant only, P < 0.001). The potential for using these antigens in a recombinant vaccine is discussed.