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Genes 2019, 10(4), 300; https://doi.org/10.3390/genes10040300

Molecular Characterization of a Dirofilaria immitis Cysteine Protease Inhibitor (Cystatin) and Its Possible Role in Filarial Immune Evasion

1
Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China
2
Department of Chemistry, College of Life and Basic Science, Sichuan Agricultural University, Chengdu 611130, China
*
Author to whom correspondence should be addressed.
These authors contributed equally to this work.
Received: 17 February 2019 / Revised: 7 April 2019 / Accepted: 8 April 2019 / Published: 12 April 2019
(This article belongs to the Section Microbial Genetics and Genomics)
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Abstract

Infection with canine heartworm (Dirofilaria immitis), spread via mosquito vectors, causes coughing, asthma, pneumonia, and bronchitis in humans and other animals. The disease is especially severe and often fatal in dogs and represents a serious threat to public health worldwide. Cysteine protease inhibitors (CPIs), also known as cystatins, are major immunomodulators of the host immune response during nematode infections. Herein, we cloned and expressed the cystatin Di-CPI from D. immitis. Sequence analysis revealed two specific cystatin-like domains, a Q-x-V-x-G motif, and a SND motif. Phylogenetic analysis indicates that Di-CPI is a member of the second subgroup of nematode type II cystatins. Probing of D. immitis total proteins with anti-rDi-CPI polyclonal antibody revealed a weak signal, and immunofluorescence-based histochemical analysis showed that native Di-CPI is mainly localized in the cuticle of male and female worms and the gut of male worms. Treatment of canine peripheral blood mononuclear cells (PMBCs) with recombinant Di-CPI induced a Th2-type immune response characterized by high expression of the anti-inflammatory factor interleukin-10. Proliferation assays showed that Di-CPI inhibits the proliferation of canine PMBCs by 15%. Together, the results indicate that Di-CPI might be related to cellular hyporesponsiveness in dirofilariasis and may help D. immitis to evade the host immune system. View Full-Text
Keywords: Dirofilaria immitis; cystatin; immunohistochemistry; peripheral blood mononuclear cells; immune evasion Dirofilaria immitis; cystatin; immunohistochemistry; peripheral blood mononuclear cells; immune evasion
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Dong, X.; Xu, J.; Song, H.; Liu, Y.; Wu, M.; Zhang, H.; Jing, B.; Lai, W.; Gu, X.; Xie, Y.; Peng, X.; Yang, G. Molecular Characterization of a Dirofilaria immitis Cysteine Protease Inhibitor (Cystatin) and Its Possible Role in Filarial Immune Evasion. Genes 2019, 10, 300.

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