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Identification of the Babesia-responsive leucine-rich repeat domain-containing protein from the hard tick Haemaphysalis longicornis

Abstract

Haemaphysalis longicornis is a tick known for transmitting Babesia parasites, including Babesia gibsoni, in East Asian countries. The vector tick must have strategies to control Babesia parasites, while Babesia parasites are also considered to establish an evasive mechanism from the tick’s innate immunity. Due to this mutual tolerance, H. longicornis is considered to be a vector of Babesia parasites. Recent studies have shown the important roles of leucine-rich repeat (LRR) domain-containing proteins in innate immunity in many living organisms. Some LRR domain-containing proteins were identified in ticks; however, their functions are still unknown. In this study, a novel LRR domain-containing protein was identified from H. longicornis (HlLRR). HlLRR contains two LRR domains, and the expression levels of mRNA and proteins were upregulated during blood feeding, particularly in the salivary glands and midgut. In addition, recombinant HlLRR (rHlLRR) demonstrated growth inhibition activity against B. gibsoni in vitro without a hemolytic effect at any concentration used. Moreover, the diameters of Babesia merozoites treated with rHlLRR were significantly larger than those of the control group. These results strongly indicate the key roles of HlLRR in the tick’s innate immunity against Babesia parasites. Furthermore, HlLRR might be a potential alternative drug to treat babesiosis.

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Acknowledgments

We are grateful to Dr. A. Iguchi of the National Research Center for Protozoan Diseases, Obihiro University of Agriculture and Veterinary Medicine, and Dr. A. Matsuu of the Transboundary Animal Diseases Research Center, Joint Faculty of Veterinary Medicine, Kagoshima University, for providing B. gibsoni (Aomori strain). We thank to Dr. H. Izumi of the Shin Nippon Biomedical Laboratories, Ltd. for providing a dog for supplying dog blood. We also thank Dr. T. Masatani of the Transboundary Animal Diseases Research Center, Joint Faculty of Veterinary Medicine, Kagoshima University, for his helpful comments on this work. This work was supported by the Japan Society for the Promotion of Science (JSPS) KAKENHI Grant Numbers 25292173, 26660229, and 26+5872. H. Maeda is supported by a Grant-in-Aid for JSPS fellows.

Author information

Correspondence to Tetsuya Tanaka.

Additional information

Hiroki Maeda and Koshi Kurisu contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Fig. 1

Confirmation of gene silencing after dsRNA injection by semiquantitative RT-PCR (PDF 17 kb)

Supplementary Fig. 2

Hemolysis level of rHlLRR and rHlLRR-ND in canine RBC. recombinant protein concentrations of 5 μM and lower were assessed. Each percentage represents the ratio vs. Triton-X as 100 % hemolysis. PBS was also used for 0 % hemolysis (PDF 11 kb)

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Maeda, H., Kurisu, K., Miyata, T. et al. Identification of the Babesia-responsive leucine-rich repeat domain-containing protein from the hard tick Haemaphysalis longicornis . Parasitol Res 114, 1793–1802 (2015). https://doi.org/10.1007/s00436-015-4365-7

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Keywords

  • Leucine-rich repeat
  • Tick
  • Babesia
  • Recombinant
  • Parasite