Screening and analysis of immune-related genes of Aedes aegypti infected with DENV2

Abstract

Dengue virus type Ⅱ (DENV2) is a primary serotype responsible for the dengue fever epidemic, and Aedes aegypti is the main DENV2 vector. Understanding the Aedes aegypti immune mechanism against DENV2 is the basis for research on immune blockade in mosquitoes. Some preliminary studies lack validation in the literature, so this study was performed to further study and validate the potential target genes to provide a further basis for screening key target genes. We screened 51 genes possibly related to Aedes aegypti infection and immunity from the literature for further verification. First, bioinformatic methods such as GO, KEGG and PPI analysis were used, and then RT-qPCR was used to detect the changes in mRNA expression in the midguts and salivary glands of Aedes aegypti infected with DENV2.Bioinformatic analysis showed that mostly genes of the glucose metabolism pathway and myoprotein were influenced. In salivary glands, the Gst (x_a) and Toll (x_b) expression levels were significantly correlated with DENV2 load (y, lg[DENV2 RNA copies]), y = -3436x_a+0.2287x_b+3.8194 (adjusted R² = 0.5563, F = 9.148, P_F = 0.0045). In midguts, DENV2 load was significantly correlated with the relative Fba(R² = 0.4381, t = 2.497, p < 0.05, df = 8), UcCr(R² = 0.4072, t = 2.344, p < 0.05, df = 8) and Gbps1(R² = 0.4678, t = 2.652, p < 0.05, df = 8) expression levels, but multiple regression did not yield significant results. This study shows that genes related to glucose metabolism and muscle proteins contribute to the interaction between Aedes aegypti and dengue virus. It was confirmed that SAAG-4, histone H4, endoplasmin, catalase and other genes are involved in the regulation of DENV2 infection in Aedes aegypti. It was revealed that GST and Toll in salivary glands may have antagonistic effects on the regulation of DENV2 load. Fba, UcCr and Gbps1 in the midgut may increase DENV2 load. These study results further condensed the potential target gene range of the Aedes aegypti immune mechanism against DENV2 infection and provided basic information for research on the Aedes aegypti in vivo blockade strategy against DENV2.

Keywords: Aedes aegypti; DENV2; Midgut; Salivary gland.