Imamura Saiki, da Silva Vaz Itabajara, Konnai Satoru, Yamada Shinji, Nakajima Chie, Onuma Misao, Ohashi Kazuhiko
Department of Disease Control, Graduate School of Veterinary Medicine, Hokkaido University, Kita 18, Nishi 9, Sapporo 060-0818, Hokkaido, Japan.
Exp Appl Acarol. 2009 Aug;48(4):345-58. doi: 10.1007/s10493-009-9245-3. Epub 2009 Jan 29.
We report the cloning, expression and characterization of an Haemaphysalis longicornis metalloprotease (named HLMP1). The gene encodes a predicted 550 aminoacid protein with similarity to metalloproteases of the reprolysin family. The protein sequence contains a signal sequence, the zinc-binding motif (HEXXHXXGXXH) common to metalloproteases and a cysteine-rich region. Reverse transcription-PCR expression analysis indicates the presence of mRNA in the salivary gland of larva, nymph and adult ticks. Rabbit repeatedly infested with H. longicornis recognized rHLMP1, suggesting that the immune-response against HLMP1 is naturally induced through the feeding of ticks. Vaccination of rabbit with rHLMP1 produced protective immunity against ticks, resulting in 15.6 and 14.6% mortality in nymph and adult ticks, respectively. This work provides information to understand the tick's defense system, and offers new insights to develop strategies to block this defense system with an anti-tick vaccine based on a metalloprotease.
我们报道了长角血蜱金属蛋白酶(命名为HLMP1)的克隆、表达及特性分析。该基因编码一个预测的含550个氨基酸的蛋白质,与解聚素家族的金属蛋白酶具有相似性。蛋白质序列包含一个信号序列、金属蛋白酶共有的锌结合基序(HEXXHXXGXXH)和一个富含半胱氨酸的区域。逆转录-聚合酶链反应表达分析表明,在幼虫、若虫和成年蜱的唾液腺中存在mRNA。反复感染长角血蜱的兔子可识别重组HLMP1,这表明针对HLMP1的免疫反应是通过蜱的取食自然诱导产生的。用重组HLMP1对兔子进行疫苗接种可产生针对蜱的保护性免疫,导致若虫和成年蜱的死亡率分别为15.6%和14.6%。这项工作为理解蜱的防御系统提供了信息,并为基于金属蛋白酶开发抗蜱疫苗来阻断该防御系统的策略提供了新的见解。