Modulation of goat monocyte function by HCcyst-2, a secreted cystatin from Haemonchus contortus
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Yujian Wang1, Yuling Wen1, Shuai Wang1, Muhammad Ehsan1, RuoFeng Yan1, XiaoKai Song1, LiXin Xu1 and XiangRui Li1
1College of Veterinary Medicine, Nanjing Agricultural University, Nanjing 210095, PR China
XiangRui Li, email: [email protected]
Keywords: Haemonchus contortus, cystatin, monocyte, immunomodulation
Received: December 23, 2016 Accepted: March 31, 2017 Published: April 21, 2017
Modulation and suppression of the host immune response by nematode parasites have been reported extensively and the cysteine protease inhibitor (cystatin) is identified as one of the major immunomodulator. In the present study, we cloned and produced recombinant cystatin protein from nematode parasite Haemonchus contortus (rHCcyst-2) and investigated its immunomodulatory effects on goat monocyte. rHCcyst-2 protein is biologically functional as shown by its ability to inhibit the protease activity of cathepsin L, cathepsin B and papain. Immunohistochemical test demonstrated that the native HCcyst-2 protein was predominantly localized at the body surface and internal surface of the worm’s gut. We demonstrated that rHCcyst-2 could be distinguished by antisera from goats experimentally infected with H. contortus and could uptake by goat monocytes. The immunomodulatory effects of HCcyst-2 on cytokine secretion, MHC molecule expression, NO production and phagocytosis were observed by co-incubation of rHCcyst-2 with goat monocytes. The results showed that the interaction of rHCcyst-2 decreased the production of TNF-α, IL-1β and IL-12p40. However, it significantly increased the secretion of IL-10 in goat monocytes. After rHCcyst-2 exposure, the expression of MHC-II on goat monocytes was inhibited. Moreover, rHCcyst-2 could up-regulate the LPS induced NO production of goat monocytes. Phagocytotic assay by FITC-dextran internalization showed that rHCcyst-2 inhibited the phagocytosis of goat monocytes. Our findings provided potential target as immunoregulator, and will be helpful to illustrate the molecular basis of host–parasite interactions and search for new potential molecule as vaccine and drug target candidate.
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