Comprehensive genetic dissection of the hemocyte immune response in the malaria mosquito anopheles gambiae

Autor: Fotis C. Kafatos, Fabrizio Lombardo, Yasmeen Ghani, George K. Christophides
Přispěvatelé: Biotechnology and Biological Sciences Research Council (BBSRC)
Jazyk: angličtina
Rok vydání: 2012
Předmět:
Hemocytes
double-stranded
Anopheles gambiae
active: genetics
Complement System
Gene Expression
active
oocysts: cytology
anopheles gambiae: immunology
0302 clinical medicine
RNA interference
1108 Medical Microbiology
cells
genome-wide association study
anopheles gambiae
phagocytosis: physiology
gene silencing
hemocytes
cell survival
hemocytes: cytology
rna
complement system proteins: immunology
immunity
oocysts: immunology
oocysts
anopheles gambiae: genetics
animals
double-stranded: pharmacology
hemocytes: immunology
insect proteins
host-pathogen interactions
phagocytosis
complement system proteins
small interfering
escherichia coli
rna interference
gene expression
cultured
hemocytes: microbiology
complement system proteins: genetics
Molecular Cell Biology
Biology (General)
GENOME-WIDE ANALYSIS
RNA
Small Interfering
Cells
Cultured
Phagosome
0303 health sciences
Cell Death
Innate Immunity
3. Good health
Cell biology
Host-Pathogen Interaction
RNA silencing
Immunity
Active
1107 Immunology
Host-Pathogen Interactions
Insect Proteins
RNA Interference
Life Sciences & Biomedicine
Research Article
0605 Microbiology
QH301-705.5
Cell Survival
Immunology
Biology
CULTURED-CELLS
Microbiology
Vector Biology
PHAGOCYTOSIS
Immune Activation
SIGNALING PATHWAYS
03 medical and health sciences
RNAI SCREEN
Immune system
STAT PATHWAY
Immunity
Virology
parasitic diseases
Anopheles
Genetics
Escherichia coli
Gene silencing
Animals
PLASMODIUM
COMPLEMENT-LIKE PROTEIN
Gene Silencing
Gene Networks
Molecular Biology
030304 developmental biology
RNA
Double-Stranded
Science & Technology
BACTERIAL CHALLENGE
Oocysts
Immune Defense
Complement System Proteins
RC581-607
biology.organism_classification
Reverse genetics
DROSOPHILA-MELANOGASTER
Immune System
Parasitology
Immunologic diseases. Allergy
Gene Function
030217 neurology & neurosurgery
Genome-Wide Association Study
Zdroj: PLoS Pathogens
PLoS Pathogens, Vol 9, Iss 1, p e1003145 (2013)
Popis: 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.
Author Summary The mosquito immune system relies on innate humoral and cellular reactions to fight infections, including those by malaria parasites that must pass through mosquitoes before they can infect humans. Therefore, a detailed molecular understanding of these reactions could assist the design of new ways to control the spread of malaria and other mosquito-borne diseases. Here we use a technique to silence in mosquito cultured cells genes that are highly and/or specifically expressed in mosquito hemocytes, the equivalent of human white blood cells, as a means to investigate their function in reactions of the mosquito immune system. Our study identifies several novel regulators of immune reactions including phagocytosis, the engulfment and subsequent destruction of bacteria and other pathogens by hemocytes, the production of antimicrobial peptides, which directly kill or inhibit the proliferation of microbes, and the basal and induced production of an important complement regulator. Complement is a robust reaction of mosquitoes against malaria parasites and bacteria through phagocytosis, lysis or melanization (the enclosure of pathogens in a melanin capsule). We also reveal intriguing molecular connections between these reactions such as phagocytosis and regulation of complement. Our study provides novel insights into mosquito immune system and its reactions against infections.
Databáze: OpenAIRE
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