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. 2019 Jun 27;20(13):3133.
doi: 10.3390/ijms20133133.

Transcriptomic Analysis of Aedes aegypti Innate Immune System in Response to Ingestion of Chikungunya Virus

Liming Zhao  1 , Barry W Alto  2 , Yongxing Jiang  3 , Fahong Yu  4 , Yanping Zhang  4
Affiliations

Transcriptomic Analysis of Aedes aegypti Innate Immune System in Response to Ingestion of Chikungunya Virus

Liming Zhao et al. Int J Mol Sci. .

Abstract

Aedes aegypti (L.) is the primary vector of emergent mosquito-borne viruses, including chikungunya, dengue, yellow fever, and Zika viruses. To understand how these viruses interact with their mosquito vectors, an analysis of the innate immune system response was conducted. The innate immune system is a conserved evolutionary defense strategy and is the dominant immune system response found in invertebrates and vertebrates, as well as plants. RNA-sequencing analysis was performed to compare target transcriptomes of two Florida Ae. aegypti strains in response to chikungunya virus infection. We analyzed a strain collected from a field population in Key West, Florida, and a laboratory strain originating from Orlando. A total of 1835 transcripts were significantly expressed at different levels between the two Florida strains of Ae. aegypti. Gene Ontology analysis placed these genes into 12 categories of biological processes, including 856 transcripts (up/down regulated) with more than 1.8-fold (p-adj (p-adjust value) ≤ 0.01). Transcriptomic analysis and q-PCR data indicated that the members of the AaeCECH genes are important for chikungunya infection response in Ae. aegypti. These immune-related enzymes that the chikungunya virus infection induces may inform molecular-based strategies for interruption of arbovirus transmission by mosquitoes.

Keywords: Aedes aegypti; anti-microbial peptide; chikungunya virus; gene expression; immune responses; transcriptome.

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Conflict of interest statement

All authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Overview of the functional categories of differentially expressed (DE) transcripts in response to CHIKV infection. DE genes were determined based on statistical analysis by DESeq package. The total number of DE genes for each comparison is shown in parentheses in each figure. Gene Ontology (GO) analysis of DE genes was performed based on the database of AmiGO 2. Up, upregulated DE genes; Down, downregulated DE genes. (A) 3 h post infection KW-CHIKV compared with KW-Control, A1 Up and A2 Down; (B) 3-day post infection, KW-CHIKV compared with KW-Control, B1 Up and B2 Down; (C) 3 days post infection, KW-Control compared with OR-Control, C1 Up and C2 Down; (D) 3 days post infection, KW-CHIKV compared with OR-CHIKV, D1 Up and D2 Down; (E) 3 days post infection, E1: OR-CHIKV compared with OR-Control, E1 UP; OR-CHIKV compared with OR-Control 339 gene down regulated with p-adj ≤ 0.01, only three genes were detected in the GO, which are not shown in the figure.
Figure 2
Figure 2
AaeDEFA, AaeDEFD, AaeDEFa, AaeDNR1, AaeCECH and AaeTEP3 relative expression level fold-changes in Aedes aegypti females that ingested CHIKV infected blood. The fold change was calculated using the 2[−average ΔΔCT] method. ΔCt (Control) = Ct (AaeDEFA/ AaeDEFD/AaeDEFa /AaeDNR1/AaeCECH/AaeTEP3) − Ct (AeaActin); ΔCt (infected-CHIKV) = Ct (AaeDEFA/ AaeDEFD/AaeDEFa /AaeDNR1/AaeCECH/AaeTEP3) − Ct (AeaActin); ΔΔCt =ΔCt (infected-CHIKV) − ΔCt (Control). The 3, 24, 48, 72, 120, 168, and 240 h (hours) represented gene expression post infection with CHIKV, (A,C) KW strain female Ae. Aegypti, (B,D) Orlando strain female Ae. aegypti.
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