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Cloning and characterization of a novel gene encoding 16 kDa protein (Ac16) from Angiostrongylus cantonensis
Zheng-Yu Li1, Zhi-Yue Lv, Jie Wei
1Key Laboratory of Tropical Diseases and Control, Ministry of Education, Guangzhou, People's Republic of China.
Abstract:
Angiostrongylus cantonensis is the most common infectious cause of eosinophilic meningitis or meningoencephalitis. A novel gene (AC16) was isolated from a cDNA library of A. cantonensis fourth-stage larvae. The putative 16-kDa protein has 149 amino acids and is homologous to an immunodominant hypodermal antigen (IHA16) from Ancylostoma caninum (identities = 57%). In this paper, we cloned the gene and purified the recombinant Ac16 (rAC16) protein. Real-time quantitative PCR revealed that Ac16 was expressed significantly higher in the fourth-stage larvae and adult worms derived from rats than that in the fourth-stage larvae derived from mice. Moreover, sera from rat (permissive host) infected with A. cantonensis detected Ac16 by Western blot, while sera from infected mouse (non-permissive host) could not. The results implied that Ac16 was related to the parasitic adaptation of A. cantonensis in different hosts and non-permissive host mouse had no circulating antibody to the antigen Ac16 from A. cantonensis and thus might contribute to understanding the mechanism of parasite immune evasion. Furthermore, we evaluated the ability of Ac16 antibody diagnosing A. cantonensis infection by an indirect enzyme-linked immunosorbent assay. The results showed that the Ac16 antibody had a 79.17% sensitivity to rAC16 and 83.33% to crude adult worm antigens (CA) (P > 0.05), while the specificity to rAC16 and to CA were 95.89% and 86.30% respectively (P < 0.05), thus implying that rAc16 may constitute a putative serodiagnostic antigen for Angiostrongyliasis cantonensis.
Insights
A novel Angiostrongylus cantonensis gene, AC16, shows higher expression in permissive rat hosts and is recognized by rat antibodies, suggesting a role in parasitic adaptation and potential for diagnosing Angiostrongyliasis cantonensis.
Area of Science:
- Parasitology
- Molecular Biology
- Immunology
Background:
- Angiostrongylus cantonensis is a significant pathogen causing eosinophilic meningitis.
- Understanding host-parasite interactions and diagnostic markers is crucial for managing Angiostrongyliasis cantonensis.
- The AC16 gene and its protein product represent a novel target for investigation.
Purpose of the Study:
- To clone and characterize the novel Angiostrongylus cantonensis gene AC16.
- To investigate the expression patterns of Ac16 in different hosts and parasitic stages.
- To evaluate the diagnostic potential of the Ac16 antigen for Angiostrongyliasis cantonensis.
Main Methods:
- Gene cloning and recombinant protein purification (rAC16).
- Real-time quantitative PCR for gene expression analysis.
- Western blot analysis using sera from infected rats and mice.
- Indirect enzyme-linked immunosorbent assay (ELISA) for diagnostic evaluation.
Main Results:
- The AC16 gene encodes a 16-kDa protein homologous to Ancylostoma caninum IHA16.
- Ac16 expression was significantly higher in fourth-stage larvae and adult worms from rats compared to mice.
- Rat sera recognized rAC16, while mouse sera did not, indicating differential host immune response.
- The Ac16 antibody demonstrated good sensitivity (79.17-83.33%) and specificity (86.30-95.89%) in ELISA.
Conclusions:
- Ac16 plays a role in the parasitic adaptation of Angiostrongylus cantonensis in different hosts.
- The lack of antibody response in mice suggests a mechanism of immune evasion.
- Recombinant Ac16 (rAC16) shows promise as a serodiagnostic antigen for Angiostrongyliasis cantonensis.
