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在宿主卵巢中对注入长角血蜱雌蜱的分泌型铁蛋白的宿主抗体进行免疫荧光检测。

Immunofluorescent detection in the ovary of host antibodies against a secretory ferritin injected into female Haemaphysalis longicornis ticks.

作者信息

Galay Remil Linggatong, Matsuo Tomohide, Hernandez Emmanuel Pacia, Talactac Melbourne Rio, Kusakisako Kodai, Umemiya-Shirafuji Rika, Mochizuki Masami, Fujisaki Kozo, Tanaka Tetsuya

机构信息

Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, Los Baños, Laguna 4031, Philippines; Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

Laboratory of Parasitology, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24 Korimoto, Kagoshima 890-0065, Japan.

出版信息

Parasitol Int. 2018 Apr;67(2):119-122. doi: 10.1016/j.parint.2017.10.006. Epub 2017 Oct 19.

DOI:10.1016/j.parint.2017.10.006
PMID:29056475
Abstract

Due to the continuous threat of ticks and tick-borne diseases to human and animal health worldwide, and the drawbacks of chemical acaricide application, many researchers are exploring vaccination as an alternative tick control method. Earlier studies have shown that host antibodies can circulate in the ticks, but it has not been confirmed whether these antibodies can be passed on to the eggs. We previously reported that ticks infesting rabbits immunized with a recombinant secretory ferritin of Haemaphysalis longicornis (HlFER2) had reduced egg production and hatching. Here we attempted to detect the presence of antibodies against HlFER2 in the ovary and eggs of female ticks through immunofluorescent visualization. Purified anti-HlFER2 antibodies or rabbit IgG for control was directly injected to engorged female H. longicornis. Ovaries and eggs after oviposition were collected and prepared for an indirect immunofluorescent antibody test. Positive fluorescence was detected in ovaries one day post-injection of anti-HlFER2 antibodies. Through silencing of Hlfer2 gene, we also determined whether the injected antibodies can specifically bind to native HlFER2. Immunofluorescence was observed in the oocytes of dsLuciferase control ticks injected with anti-HlFER2 antibodies, but not in the oocytes of Hlfer2-silenced ticks also injected with anti-HlFER2 antibodies. Our current findings suggest that host antibodies can be passed on to the oocytes, which is significant in formulating a vaccine that can disrupt tick reproduction.

摘要

由于蜱虫和蜱传疾病在全球范围内对人类和动物健康构成持续威胁,以及化学杀螨剂应用存在的弊端,许多研究人员正在探索将疫苗接种作为一种替代的蜱虫控制方法。早期研究表明,宿主抗体可在蜱虫体内循环,但这些抗体是否能传递给卵尚未得到证实。我们之前报道,用长角血蜱重组分泌型铁蛋白(HlFER2)免疫的兔子身上寄生的蜱虫产卵量和孵化率降低。在此,我们试图通过免疫荧光可视化检测雌性蜱虫卵巢和卵中抗HlFER2抗体的存在。将纯化的抗HlFER2抗体或作为对照的兔IgG直接注射到饱血的雌性长角血蜱体内。收集产卵后的卵巢和卵,准备进行间接免疫荧光抗体试验。注射抗HlFER2抗体后一天,在卵巢中检测到阳性荧光。通过沉默Hlfer2基因,我们还确定了注射的抗体是否能特异性结合天然HlFER2。在注射抗HlFER2抗体的dsLuciferase对照蜱虫的卵母细胞中观察到免疫荧光,但在同样注射抗HlFER2抗体的Hlfer2沉默蜱虫的卵母细胞中未观察到。我们目前的研究结果表明,宿主抗体可以传递给卵母细胞,这对于制定一种能够干扰蜱虫繁殖的疫苗具有重要意义。

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