Comparative analysis for immune response of coelomic fluid from coelom and polian vesicle in Apostichopus japonicus to Vibrio splendidus infection

The polian vesicle and coelom of sea cucumber Apostichopus japonicus were full of coelomic fluid in which many types of coelomocytes with different functions were suspended. Our previous work has indicated the differences of coelomocytes between two sites mainly in subtype proportion, non-specific immune enzymes activities and several immune-related genes expression levels in healthy A. japonicus. However, the functional similarities and differences of coelomic fluid in two sites including the coelom and polian vesicle after pathogenic infection still remain unclear. Here, we investigated the changes of the total coelomocyte density (TCD) and differential coelomocyte density (DCD) after pathogen infection by Vibrio splendidus in coelom and polian vesicle. After infected by V. splendidus, the TCD in the coelom and polian vesicle rapidly declined at 12 h, and then the TCD in the coelom showed a stably ascending trend, while the TCD in the polian vesicle reached a peak at 24 h post infection (hpi), and then showed a continuously decline trend from 24 hpi to 72 hpi followed by a slow elevation until recovering the normal level from 72 hpi to 96 hpi. Then the activities of acidic phosphatase (ACP), alkaline phosphatase (AKP), catalase (CAT) and superoxide dismutase (SOD) were determined to evaluate the response of cell-free coelomic fluid to V. splendidus infection. The activities of ACP, AKP and CAT showed similar trends in the coelom and polian vesicle. The SOD activity significantly increased in the polian vesicle, whereas it exhibited a decreasing trend in the coelom. Finally, the expression profiles of nine immune-related genes including AjMyD88, Aj-IRAK4, Aj-i-Lys, Aj-Rel, Aj-p50, Aj-DMBT1, Aj-CDC, Aj-Rrp15 and Aj-Fibrinogen C were detected after V. splendidus challenge. The results suggested all the detected genes were significantly up-regulated both in the coelom and polian vesicle, and the expression levels of these genes in two sites shared similar trends except AjMyD88 and Aj-DMBT1. This research provides a new insight into the differentially immune roles of coelomic fluid and coelomocytes in polian vesicle and coelom response to bacterial infections and supplements comprehensive resources for better understanding the innate immune response of A. japonicus.