title: Characterization of the Rel2-regulated transcriptome and proteome of Anopheles stephensi identifies new anti-Plasmodium factors.
abstract: Mosquitoes possess an innate immune system that is capable of limiting infection by a variety of pathogens, including the Plasmodium spp. parasites responsible for human malaria. The Anopheles immune deficiency (IMD) innate immune signaling pathway confers resistance to Plasmodium falciparum. While some previously identified Anopheles anti-Plasmodium effectors are regulated through signaling by Rel2, the transcription factor of the IMD pathway, many components of this defense system remain uncharacterized. To begin to better understand the regulation of immune effector proteins by the IMD pathway, we used oligonucleotide microarrays and iTRAQ to analyze differences in mRNA and protein expression, respectively, between transgenic Anopheles stephensi mosquitoes exhibiting blood meal-inducible overexpression of an active recombinant Rel2 and their wild-type conspecifics. Numerous genes were differentially regulated at both the mRNA and protein levels following induction of Rel2. While multiple immune genes were up-regulated, a majority of the differentially expressed genes have no known immune function in mosquitoes. Selected up-regulated genes from multiple functional categories were tested for both anti-Plasmodium and anti-bacterial action using RNA interference (RNAi). Based on our experimental findings, we conclude that increased expression of the IMD immune pathway-controlled transcription factor Rel2 affects the expression of numerous genes with diverse functions, suggesting a broader physiological impact of immune activation and possible functional versatility of Rel2. Our study has also identified multiple novel genes implicated in anti-Plasmodium defense.
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205-2179, USA. Electronic address: apike@jhsph.edu.
authors.forename: Andrew
authors.initials: A
authors.lastname: Pike
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205-2179, USA. Electronic address: alvadlam@jhsph.edu.
authors.forename: Alekhya
authors.initials: A
authors.lastname: Vadlamani
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205-2179, USA. Electronic address: ssandifo@jhsph.edu.
authors.forename: Simone L
authors.initials: SL
authors.lastname: Sandiford
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205-2179, USA. Electronic address: agacita1@jhu.edu.
authors.forename: Anthony
authors.initials: A
authors.lastname: Gacita
authors.affiliation: W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Johns Hopkins Malaria Research Institute, Bloomberg School of Public Health, Johns Hopkins University, 615 N. Wolfe Street, Baltimore, MD 21205-2179, USA. Electronic address: gdimopou@jhsph.edu.
authors.forename: George
authors.initials: G
authors.lastname: Dimopoulos
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D000852:Anopheles / Q000235:genetics* / Q000276:immunology* / Q000469:parasitology*; D005260:Female; D020869:Gene Expression Profiling; D007113:Immunity, Innate; D019476:Insect Proteins / Q000096:biosynthesis; D007303:Insect Vectors / Q000235:genetics* / Q000276:immunology* / Q000469:parasitology*; D020411:Oligonucleotide Array Sequence Analysis; D010963:Plasmodium falciparum / Q000502:physiology; D020543:Proteome; D034622:RNA Interference; D015398:Signal Transduction / Q000502:physiology; D014157:Transcription Factors; D059467:Transcriptome / Q000502:physiology*; D015854:Up-Regulation
chemical_list: D019476:Insect Proteins; D020543:Proteome; D014157:Transcription Factors
keywords: Innate immunity; Mosquitoes; Plasmodium; Proteome; Rel2; Transcriptome; Transgenesis