lncRNA028466 Regulates Th1/Th2 Cytokines Expression and Associated with Echinococcus Granulosus Antigen P29 Immunity

Abstract

Background: Cystic echinococcosis (CE) is a parasite disease that was caused by Echinococcus granulosus (Eg). The recombinant Echinococcus granulosus antigen P29 (rEg.P29) has been shown to conferred effective immunity to sheep and mice during Echinococcus granulosus secondary infection in our previous study. In our study, we strove to research the ability of Long noncoding RNA 028466 (lncRNA028466) as a regulator for the protective immunity mediated by rEg.P29 vaccination and studied the effects of lncRNA028466 on CD4+T cells differentiation in mice spleen.Methods: Female BALB/c mice were divided into two groups and were vaccinated subcutaneously with rEg.P29 antigen and PBS as a control (12 mice each group). Following prime-boost vaccination, CD4+T, CD8+T，and B cells from the spleen were isolated by flow cytometry. Quantitative real-time PCR (qRT-PCR) was performed to measure the expression of lncRNA028466 in these three kinds of cells. Then, lncRNA028466 was over-expressed and knock-downed. Furthermore, were d by qRT-PCR and ELISA were performed to evaluate the production of IFN-γ, IL-2, IL-4, and IL-10, as well as was flow cytometry was performed to detect the differentiation of Th1 and Th2 subgroups. Results: lncRNA028466 was significantly decreased after the second-week of immunization with rEg.P29 antigen. The proportion of CD4+T cells was increased after rEg.P29 immunization. Subsequent overexpression and knockdown of lncRNA028466 in naive CD4+T cells were performed to detect changes in Th1 and Th2 cytokines expression. Overexpression of lncRNA028466 facilitated the production of IL-4, IL-10 and suppressed the production of IFN-γ, IL-2. Furthermore, after transfection with siRNA028466, IL-2 production was facilitated and IL-10 production was suppressed in naive CD4+T cells.Conclusions: Immunization with rEg.P29 down-regulated the expression of lncRNA028466, which was related to a higher Th1 immune response, a higher Th2 immune response with lncRNA028466 overexpression, a higher IL-2 mRNA expression, and a lower IL-10 mRNA expression with lncRNA028466 knockdown. Our results suggest that lncRNA028466 may be involved in rEg.P29-mediated immune response by regulating cytokines expression of Th1 and Th2.