Comprehensive genetic dissection of the hemocyte immune response in the malaria mosquito anopheles gambiae
Author(s)
Lombardo, Fabrizio
Ghani, Yasmeen
Kafatos, Fotis C
Christophides, George K
Type
Journal Article
Abstract
Reverse genetics in the mosquito Anopheles gambiae by RNAi mediated gene silencing has led in recent years to an advanced understanding of the mosquito immune response against infections with bacteria and malaria parasites. We developed RNAi screens in An. gambiae hemocyte-like cells using a library of double-stranded RNAs targeting 109 genes expressed highly or specifically in mosquito hemocytes to identify novel regulators of the hemocyte immune response. Assays included phagocytosis of bacterial bioparticles, expression of the antimicrobial peptide CEC1, and basal and induced expression of the mosquito complement factor LRIM1. A cell viability screen was also carried out to assess dsRNA cytotoxicity and to identify genes involved in cell growth and survival. Our results identify 22 novel immune regulators, including proteins putatively involved in phagosome assembly and maturation (Ca2+ channel, v-ATPase and cyclin-dependent protein kinase), pattern recognition (fibrinogen-domain lectins and Nimrod), immune modulation (peptidase and serine protease homolog), immune signaling (Eiger and LPS-induced factor), cell adhesion and communication (Laminin B1 and Ninjurin) and immune homeostasis (Lipophorin receptor). The development of robust functional cell-based assays paves the way for genome-wide functional screens to study the mosquito immune response to infections with human pathogens.
Date Issued
2013-01-01
Date Acceptance
2012-12-05
Citation
PLoS Pathogens, 2013, 9 (1), pp.1-11
ISSN
1553-7366
Publisher
Public Library of Science (PLoS)
Start Page
1
End Page
11
Journal / Book Title
PLoS Pathogens
Volume
9
Issue
1
Copyright Statement
© 2013 Lombardo et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
License URL
Sponsor
Biotechnology and Biological Sciences Research Council (BBSRC)
Identifier
http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcApp=PARTNER_APP&SrcAuth=LinksAMR&KeyUT=WOS:000314464600048&DestLinkType=FullRecord&DestApp=ALL_WOS&UsrCustomerID=1ba7043ffcc86c417c072aa74d649202
Grant Number
BB/E002641/1
Subjects
Science & Technology
Life Sciences & Biomedicine
Microbiology
Parasitology
Virology
COMPLEMENT-LIKE PROTEIN
GENOME-WIDE ANALYSIS
RNAI SCREEN
DROSOPHILA-MELANOGASTER
BACTERIAL CHALLENGE
SIGNALING PATHWAYS
CULTURED-CELLS
STAT PATHWAY
PLASMODIUM
PHAGOCYTOSIS
Publication Status
Published
Article Number
ARTN e1003145
Date Publish Online
2013-01-31