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Expression and biological functions of Ancylostoma ceylanicum saposin-like protein

  • Helminthology - Original Paper
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Abstract

Ancylostoma ceylanicum is a common zoonotic nematode that inhabits the small intestine of humans, dogs, and cats. Saposin-like proteins (SLPs) have hemolytic and antibacterial activities and could be used as diagnostic or vaccine candidates. To explore the biological functions of Ancylostoma ceylanicum SLP (Ace-SLP-1), cDNA-encoding Ace-SLP-1 mature peptide was cloned into prokaryotic expression vector pET-28a and transformed into Escherichia coli BL21 (DE3) to induce expression. After incubation of canine red blood cell suspension with different concentrations of recombinant Ace-SLP-1, the supernatant was separated to measure OD value and calculate the hemolysis rate. The different concentrations of recombinant protein were co-cultured with E. coli and Enterococcus faecalis, and colony-forming units (CFU) were determined by the plate counting method. Peripheral blood mononuclear cells (PBMCs) from healthy dogs were incubated with different concentrations of recombinant Ace-SLP-1, and the cytokine expression was evaluated by relative quantitative PCR. Our results showed that the hemolytic activity of Ace-SLP-1 increased with the increase in protein concentration from 25 to 100 μg/mL. The recombinant protein had no antibacterial activity against the two kinds of bacteria but could stimulate the secretion of cytokines (IL-4, IL-10, IL-12, and IL-13) in canine PBMCs. These data suggest that Ace-SLP-1 is involved in hookworm blood-feeding and survival and has good immunogenicity, supporting its potential as a diagnostic and vaccine target molecule.

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Funding

This work was supported by grants from the National Natural Science Foundation of China (Grant No. 31672541) and the Science and Technology Planning Project of Guangdong Province, China (Grant No. 2014A020214005).

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Authors and Affiliations

  1. Guangdong Provincial Key Laboratory of Zoonosis Prevention and Control, College of Veterinary Medicine, South China Agricultural University, Guangzhou, 510542, China

    Long He, Asmaa M. I. Abuzeid, Tingting Zhuang, Qi Zhao, Shilan Zhu, Xiaoyu Chen, Jumei Liu, Xiu Li & Guoqing Li

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  1. Long He
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  2. Asmaa M. I. Abuzeid
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  3. Tingting Zhuang
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  4. Qi Zhao
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  5. Shilan Zhu
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  7. Jumei Liu
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  8. Xiu Li
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  9. Guoqing Li
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Correspondence to Guoqing Li.

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Ethical approval

Animal experiments involved in this study were performed in accordance with the guidelines of the Animal Welfare Law and Regulations of the Department of Health and Human Services, China, and were reviewed and approved by the South China Agricultural University Animal Care and Use Committee (approval number SYXK(Yue)2019–0136).

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The authors declare no competing interests.

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He, L., Abuzeid, A.M.I., Zhuang, T. et al. Expression and biological functions of Ancylostoma ceylanicum saposin-like protein. Parasitol Res 120, 3805–3813 (2021). https://doi.org/10.1007/s00436-021-07313-6

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  • DOI: https://doi.org/10.1007/s00436-021-07313-6

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