Molecular Profiling of Phagocytic Immune Cells in Anopheles gambiae Reveals Integral Roles for Hemocytes in Mosquito Innate Immunity.
The innate immune response is highly conserved across all eukaryotes and has been studied in great detail in several model organisms. Hemocytes, the primary immune cell population in mosquitoes, are important components of the mosquito innate immune response, yet critical aspects of their biology have remained uncharacterized. Using a novel method of enrichment, we isolated phagocytic granulocytes and quantified their proteomes by mass spectrometry. The data demonstrate that phagocytosis, blood-feeding, and Plasmodium falciparum infection promote dramatic shifts in the proteomic profiles of An. gambiae granulocyte populations. Of interest, large numbers of immune proteins were induced in response to blood feeding alone, suggesting that granulocytes have an integral role in priming the mosquito immune system for pathogen challenge. In addition, we identify several granulocyte proteins with putative roles as membrane receptors, cell signaling, or immune components that when silenced, have either positive or negative effects on malaria parasite survival. Integrating existing hemocyte transcriptional profiles, we also compare differences in hemocyte transcript and protein expression to provide new insight into hemocyte gene regulation and discuss the potential that post-transcriptional regulation may be an important component of hemocyte gene expression. These data represent a significant advancement in mosquito hemocyte biology, providing the first comprehensive proteomic profiling of mosquito phagocytic granulocytes during homeostasis blood-feeding, and pathogen challenge. Together, these findings extend current knowledge to further illustrate the importance of hemocytes in shaping mosquito innate immunity and their principal role in defining malaria parasite survival in the mosquito host.
From the ‡W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, Maryland 21205.|**Department of Entomology, Iowa State University, Ames, Iowa 50011.
Ryan C
RC
Smith
From the ‡W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, Maryland 21205.|‡‡Department of Biochemistry, Molecular Biology, Entomology, and Plant Pathology, Mississippi State University, Starkville, Mississippi 39762.
Jonas G
JG
King
From the ‡W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, Maryland 21205.|§§Division of Pre-clinical Innovation, National Center for Advancing Translational Sciences, National Institutes of Health, Rockville, Maryland 20850.
Dingyin
D
Tao
§Bioinformatics, Institute for Knowledge Discovery, Graz University of Technology, 8010 Graz, Austria.|¶Core Facility Bioinformatics, Austrian Centre of Industrial Biotechnology, 8010 Graz, Austria.|‖BioTechMed OMICS Center Graz, 8010 Graz, Austria.
Oana A
OA
Zeleznik
From the ‡W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, Maryland 21205.
Clara
C
Brando
§Bioinformatics, Institute for Knowledge Discovery, Graz University of Technology, 8010 Graz, Austria.|¶Core Facility Bioinformatics, Austrian Centre of Industrial Biotechnology, 8010 Graz, Austria.|‖BioTechMed OMICS Center Graz, 8010 Graz, Austria.
Gerhard G
GG
Thallinger
From the ‡W. Harry Feinstone Department of Molecular Microbiology and Immunology and the Malaria Research Institute, Johns Hopkins Bloomberg School of Public Health, 615 North Wolfe Street, Baltimore, Maryland 21205; rdinglasan@epi.ufl.edu.|¶¶Emerging Pathogens Institute, Department of Infectious Diseases & Immunology, University of Florida, Gainesville, Florida 32611.
Rhoel R
RR
Dinglasan
D000818:Animals; D000852:Anopheles / Q000276:immunology* / Q000378:metabolism / Q000469:parasitology; D005260:Female; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D006434:Hemocytes / Q000378:metabolism*; D007113:Immunity, Innate; D019476:Insect Proteins / Q000235:genetics / Q000378:metabolism*; D013058:Mass Spectrometry; D010587:Phagocytosis; D010963:Plasmodium falciparum / Q000276:immunology; D040901:Proteomics / Q000379:methods*; D013312:Stress, Physiological
D019476:Insect Proteins