Identification of putative innate immune related genes from a cell line of the mosquito Aedes albopictus following bacterial challenge

Abstract

We report identification of putative innate immune related genes from a cell line of the mosquito Aedes albopictus challenged with heat-killed bacteria. Using a subtractive hybridization and sequencing approach, we analyzed a total 309 expressed sequence tags (ESTs) which clustered in 40 contigs. Thirty-five percent of genes yielded homology to known immune genes corresponding to antimicrobial peptides (AMPs), pathogen-associated molecular patterns, protease and immune signaling cascades. Interestingly, most of the genes have not been previously described from this mosquito and thus represent a class of novel immune genes. Further, 25% sequences did not match to any known species in the non-redundant databases, appear to be specific to the mosquito A. albopictus and merit further study.

Keywords

Aedes albopictus innate immunity subtractive hybridization mosquito ESTs

References

Medzhitov R. , Janeway CA Innate immunity: the virtues of a nonclonal system of recognition. Cell 1997; 91: 295-298.

Kim C. , Park J. , Ha N. , Kang HJ , Lee BL Innate immune response in insects: recognition of bacterial peptidoglycan and amplification of its recognition signal. BMB Reports 2008; 41: 93-101.

Barillas-Mury C. , Wizel B. , Han YS Mosquito immune responses and malaria transmission: lessons from insect model systems and implications for vertebrate innate immunity and vaccine development. Insect Biochem Mol Biol 2000; 30: 429-442.

Lowenberger C. Innate immune response of Aedes aegypti. Insect Biochem Mol Biol 2001; 31: 219-229.

Scholte JE , Schaffner F. Waiting for the tiger: establishment and spread of the Aedes albopictus mosquito in Europe. In: von Takken W, Knols BGJ, (eds) Emerging pests and vector-borne diseases in Europe, Vol. 1. Wageningen: Academic Press, 2007 ; 24-260.

Dixit R. , Sharma A. , Mourya DT , Kamaraju R. , Patole MS , Shouche YS Salivary gland transcriptome analysis during Plasmodium infection in malaria vector Anopheles stephensi. Int J Infect Dis 2009; 13: 636-646.

Fick Jr RB. (ed) Pseudomonas aeruginosa the Opportunist: Pathogenesis and Disease. Boca Raton, FL: CRC Press, 1993.

D’Argenio DA , Gallagher LA , Berg CA , Manoil C. Drosophila as a model host for Pseudomonas aeruginosa infection. J Bacteriol 2001; 183: 1466-1471.

Mahajan-Miklos S. , Tan MW , Rahme LG , Ausubel FM Molecular mechanisms of bacterial virulence elucidated using a Pseudomonas aeruginosa-Caenorhabditis elegans pathogenesis model. Cell 1999; 96: 47-56.

10.

Rahme LG , Stevens EJ , Wolfort SF , Shao J. , Tompkins RG , Ausubel FM Common virulence factors for bacterial pathogenicity in plants and animals . Science 1995; 268: 1899-1902.

11.

Hillyer JF , Schmidt SL , Christensen BM The antibacterial innate immune response by the mosquito Aedes aegypti is mediated by hemocytes and independent of Gram type and pathogenicity. Microbes Infect 2004; 6: 448-459.

12.

Lehmann T. , Hume JC , Licht M. et al. Molecular evolution of immune genes in the malaria mosquito Anopheles gambiae. PLoS One 2009; 4: e4549.

13.

Dimopoulos G. , Seeley D. , Wolf A. , Kafatos FC Malaria infection of the mosquito Anopheles gambiae activates immune-responsive genes during the critical stages of the parasite life cycle. EMBO J 1998; 17: 6115-6123.

14.

Cociancich S. , Ghazi A. , Hoffmann JA , Hetrus C. , Letellier C. Insect defensin, an inducible antibacterial peptide, forms voltage dependent channels in Micrococcus luteus. J Biol Chem 1993; 268: 19239-19245.

15.

Huang LC , Redfern RL , Narayanan S. , Reins RY , McDermott AM In vitro activity of human β-defensin 2 against Pseudomonas aeruginosa in the presence of tear fluid. Antimicrob Agents Chemother 2007; 51: 3853-3860.

16.

Gao Y. , Hernandez VP , Fallon AM Immunity proteins from mosquito cell lines include three defensin A isoforms from Aedes aegypti and a defensin D from Aedes albopictus. Insect Mol Biol 1999; 8: 311-318.

17.

Simard F. , Licht M. , Besansky NJ , Lehmann T. Polymorphism at the defensin gene in the Anopheles gambiae complex: testing different selection hypotheses. Infect Genet Evol 2007; 7: 285-292.

18.

Akira S. , Uematsu S. , Takeuchi O. Pathogen recognition and innate immunity. Cell 2006; 124: 783-801.

19.

Fabrick JA , Baker JE , Kanost MR Innate immunity in a Pyralid moth: functional evaluation of domains from a beta-1,3-glucan recognition protein. J Biol Chem 2004; 279: 26605-26611.

20.

Choe K. , Werner T. , Stöven S. , Hultmark D. , Anderson KV Requirement for a peptidoglycan recognition protein (PGRP) in Relish activation and antibacterial immune responses in Drosophila. Science 2002; 296: 359-362.

21.

Jiggins FM , Kim K. Contrasting evolutionary patterns in Drosophila immune receptors. J Mol Evol 2006; 63: 769-780.

22.

Dimopoulos G. , Richman A. , Müller HM , Kafatos FC Molecular immune responses of the mosquito Anopheles gambiae to bacteria and malaria parasites. Proc Natl Acad Sci USA 1997; 94: 11508-11513.

23.

Bulmer MS , Bachelet I. , Raman R. , Rosengaus RB , Sasisekharan R. Targeting an antimicrobial effector function in insect immunity as a pest control strategy. Proc Natl Acad Sci USA 2009 ; 106: 12652-12657.

24.

Sottrup- Jensen L. Alpha-macroglobulins: structure, shape, and mechanism of proteinase complex formation. J Biol Chem 1989; 264: 11539-11542.

25.

Blandin SA , Marois E. , Levashina EA Antimalarial responses in Anopheles gambiae: from a complement-like protein to a complement-like pathway. Cell Host Microbe 2008; 3: 364-374.

26.

Levashina EA , Moita LF , Blandin S. , Vriend G. , Lagueux M. , Kafatos FC Conserved role of a complement-like protein in phagocytosis revealed by dsRNA knockout in cultured cells of the mosquito, Anopheles gambiae. Cell 2001; 104: 709-718.

27.

Wang X. , Zhao Q. , Christensen BM Identification and characterization of the fibrinogen-like domain of fibrinogen-related proteins in the mosquito, Anopheles gambiae, and the fruitfly, Drosophila melanogaster, genomes. BMC Genomics 2005; 6: 114.

28.

Dixit R. , Roy U. , Patole MS , Shouche YS Molecular and phylogenetic analysis of a novel family of fibrinogen-related proteins from mosquito Aedes albopictus cell line. Comput Biol Chem 2008; 32: 382-386.

29.

Christophides GK , Zdobnov E. , Barillas-Mury C. et al. Immunity-related genes and gene families in Anopheles gambiae. Science 2002; 298: 159-165.

30.

Rodrigues J. , Agrawal N. , Sharma A. et al. Transcriptional analysis of an immune-responsive serine protease from Indian malarial vector, Anopheles culicifacies. BMC Mol Biol 2007; 15: 8-33.

31.

Ragg H. The role of serpins in the surveillance of the secretory pathway. Cell Mol Life Sci 2007; 64: 2763-2770.

32.

Abraham EG , Pinto SB , Ghosh A. et al. An immune-responsive serpin, SRPN6, mediates mosquito defense against malaria parasites. Proc Natl Acad Sci USA 2005; 102: 16327-16332.

33.

Chandra D. , Liu JW , Tang DG Early mitochondrial activation and cytochrome c up-regulation during apoptosis . J Biol Chem 2002; 277: 50842-50854.

34.

Opferman JT , Korsmeyer SJ Apoptosis in the development and maintenance of the immune system. Nat Immunol 2003; 4: 410-415.

35.

Nappi AJ , Ottaviani E. Cytotoxicity and cytotoxic molecules in invertebrates. Bioessays 2000; 22: 469-480.

36.

Greenberg S. , Grinstein S. Phagocytosis and innate immunity. Curr Opin Immunol 2002; 14: 136-145.

37.

Boutros M. , Agaisse H. , Perrimone N. Sequential activation of signaling pathways during innate immune responses in Drosophila. Dev Cell 2002; 3: 711-722.