lncRNA028466 regulates Th1/Th2 cytokine expression and associates with Echinococcus granulosus antigen P29 immunity

Abstract

Background Cystic echinococcosis (CE) is a parasitic disease that is caused by Echinococcus granulosus (Eg). The recombinant Echinococcus granulosus antigen P29 (rEg.P29) was shown to confer effective immunity to sheep and mice during E. granulosus secondary infection in our previous study. In this study, we sought to investigate the ability of long noncoding RNA 028466 (lncRNA028466) as a regulator for the protective immunity mediated by rEg.P29 vaccination and to study the effects of lncRNA028466 on CD4+T cell 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 overexpressed and knocked down in naive CD4+T cells, and Th1 and Th2 cytokine expression was detected. qRT-PCR, western blot, and ELISA were performed to evaluate the production of IFN-γ, IL-2, IL-4, and IL-10, and 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. 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 downregulated the expression of lncRNA028466, which was related to a higher Th1 immune response and a lower Th2 immune response. Our results suggest that lncRNA028466 may be involved in rEg.P29-mediated immune response by regulating cytokine expression of Th1 and Th2.

Long Noncoding RNAs as Orchestrators of CD4+ T-Cell Fate

“…Additionally, the lncRNA Dreg1 is intimately linked to the expression of GATA3 during Th2-cell differentiation and participates in the establishment of GATA3 expression ( Chan et al, 2021 ), suggesting that Dreg1 may promote Th2-cell differentiation. The expression of LncRNA028466 decreased after inoculation with recombinant Echinococcus granulosus antigen P29 vaccine, and studies have shown that LncRNA028466 can promote Th2-cell differentiation and the secretion of IL-10 and IL-4, but inhibits the secretion of Th1-related cytokines, IFN-γ and IL-2 ( Wang et al, 2021 ). Cystic echinococcosis is a chronic parasitic disease for which there is still no commercial vaccine due to the complex host-parasite relationship.…”

“…Cystic echinococcosis is a chronic parasitic disease for which there is still no commercial vaccine due to the complex host-parasite relationship. However, some effective antigens have been used as candidate vaccines for cystic echinococcosis prevention, such as recombinant Echinococcus granulosus antigen P29 ( Wang et al, 2021 ). LncRNA028466 may be involved in recombinant Echinococcus granulosus antigen P29 vaccination-mediated immune response by regulating the differentiation of CD4 + T cells during echinococcus granulosus infection ( Wang et al, 2021 ).…”

“…However, some effective antigens have been used as candidate vaccines for cystic echinococcosis prevention, such as recombinant Echinococcus granulosus antigen P29 ( Wang et al, 2021 ). LncRNA028466 may be involved in recombinant Echinococcus granulosus antigen P29 vaccination-mediated immune response by regulating the differentiation of CD4 + T cells during echinococcus granulosus infection ( Wang et al, 2021 ). Finally, various knockdown and overexpression experiments indicate that lncRNAs play a key role in CD4 + T-cell differentiation.…”

Adaptive immune responses and cytokine immune profiles in humans following prime and boost vaccination with the SARS-CoV-2 CoronaVac vaccine

“…PBMCs (5 × 10 5 ) were resuspended in 100 μL of Buffer2 (as prevous reported [ 32 ]) for the surface stain. Then, the surface markers were added to stain the cells for half-hour at 4 °C, protected from light.…”

“…Then, the surface markers were added to stain the cells for half-hour at 4 °C, protected from light. For intracellular cytokine staining, samples were then fixed for 8 min protecting from light using 4% paraformaldehyde and permeabilised for 2 h in the dark using Buffer2 (as prevous reported [ 32 ]). After washing, the cells were stained using intranuclear antibodies in the dark for half-hour at 4 °C.…”