Journal
MOLECULES
Volume 28, Issue 4, Pages -Publisher
MDPI
DOI: 10.3390/molecules28041806
Keywords
Mytilus coruscus; interleukin-17; RNA interference; flow cytometric analysis; inflammatory response
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The newly identified member of the IL17 family, McIL17-1, plays an important role in the innate immune response to bacterial infection in M. coruscus. It is expressed in various tissues with high levels in gills, hemocytes, and digestive glands. McIL17-1 is upregulated after stimulation with different pathogen associated molecular patterns (PAMPs) and has a cytoplasmic localization in HEK293T cells. Knockdown of McIL17-1 reduces mortality rate in response to Vibrio alginolyticus infection, while overexpression increases mortality rate.
The inflammatory cytokine interleukin-17 (IL17) plays an important role in innate immunity by binding to its receptors (IL17Rs) to activate immune defense signals. To date, information on members of the IL17 family is still very limited in molluscan species. Here, a novel member of the IL17 family was identified and characterized from thick shell mussel Mytilus coruscus, and this gene was designated as McIL17-1 by predicting structural domains and phylogenetic analysis. McIL17-1 transcripts existed in all examined tissues with high expression levels in gills, hemocytes and digestive glands. After the stimuli of different pathogen associated molecular patterns (PAMPs) for 72 h, transcriptional expression of McIL17-1 was significantly upregulated, except for poly I:C stimulation. Cytoplasm localization of McIL17-1 was shown in HEK293T cells by fluorescence microscopy. Further, in vivo and in vitro assays were performed to evaluate the potential function of McIL17-1 played in immune response. McIL17-1 was either knocked down or overexpressed in vivo through RNA inference (RNAi) and recombinant protein injection, respectively. With the infection of living Vibrio alginolyticus, a high mortality rate was exhibited in the McIL17-1 overexpressed group compared to the control group, while a lower mortality rate was observed in the McIL17-1 knocked down group than control group. In vitro, the flow cytometric analysis showed that the apoptosis rate of McIL17-1 inhibited hemocytes was significantly lower than that of the control group after lipopolysaccharide stimulation. These results collectively suggested that the newly identified IL17 isoform is involved in the inflammatory response to bacterial infection in M. coruscus.
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