Skip to main content
PMC home page
The EMBO Journal logoLink to The EMBO Journal
. 1998 Mar 2;17(5):1217–1227. doi: 10.1093/emboj/17.5.1217

A drosomycin-GFP reporter transgene reveals a local immune response in Drosophila that is not dependent on the Toll pathway.

D Ferrandon 1, A C Jung 1, M Criqui 1, B Lemaitre 1, S Uttenweiler-Joseph 1, L Michaut 1, J Reichhart 1, J A Hoffmann 1
PMCID: PMC1170470  PMID: 9482719

Abstract

A hallmark of the systemic antimicrobial response of Drosophila is the synthesis by the fat body of several antimicrobial peptides which are released into the hemolymph in response to a septic injury. One of these peptides, drosomycin, is active primarily against fungi. Using a drosomycin-green fluorescent protein (GFP) reporter gene, we now show that in addition to the fat body, a variety of epithelial tissues that are in direct contact with the external environment, including those of the respiratory, digestive and reproductive tracts, can express the antifungal peptide, suggesting a local response to infections affecting these barrier tissues. As is the case for vertebrate epithelia, insect epithelia appear to be more than passive physical barriers and are likely to constitute an active component of innate immunity. We also show that, in contrast to the systemic antifungal response, this local immune response is independent of the Toll pathway.

Full Text

The Full Text of this article is available as a PDF (502.8 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Asling B., Dushay M. S., Hultmark D. Identification of early genes in the Drosophila immune response by PCR-based differential display: the Attacin A gene and the evolution of attacin-like proteins. Insect Biochem Mol Biol. 1995 Apr;25(4):511–518. doi: 10.1016/0965-1748(94)00091-c. [DOI] [PubMed] [Google Scholar]
  2. Baeuerle P. A., Baltimore D. NF-kappa B: ten years after. Cell. 1996 Oct 4;87(1):13–20. doi: 10.1016/s0092-8674(00)81318-5. [DOI] [PubMed] [Google Scholar]
  3. Brey P. T., Lee W. J., Yamakawa M., Koizumi Y., Perrot S., François M., Ashida M. Role of the integument in insect immunity: epicuticular abrasion and induction of cecropin synthesis in cuticular epithelial cells. Proc Natl Acad Sci U S A. 1993 Jul 1;90(13):6275–6279. doi: 10.1073/pnas.90.13.6275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Bulet P., Dimarcq J. L., Hetru C., Lagueux M., Charlet M., Hegy G., Van Dorsselaer A., Hoffmann J. A. A novel inducible antibacterial peptide of Drosophila carries an O-glycosylated substitution. J Biol Chem. 1993 Jul 15;268(20):14893–14897. [PubMed] [Google Scholar]
  5. Chalfie M., Tu Y., Euskirchen G., Ward W. W., Prasher D. C. Green fluorescent protein as a marker for gene expression. Science. 1994 Feb 11;263(5148):802–805. doi: 10.1126/science.8303295. [DOI] [PubMed] [Google Scholar]
  6. Charlet M., Lagueux M., Reichhart J. M., Hoffmann D., Braun A., Meister M. Cloning of the gene encoding the antibacterial peptide drosocin involved in Drosophila immunity. Expression studies during the immune response. Eur J Biochem. 1996 Nov 1;241(3):699–706. doi: 10.1111/j.1432-1033.1996.00699.x. [DOI] [PubMed] [Google Scholar]
  7. Cormack B. P., Valdivia R. H., Falkow S. FACS-optimized mutants of the green fluorescent protein (GFP). Gene. 1996;173(1 Spec No):33–38. doi: 10.1016/0378-1119(95)00685-0. [DOI] [PubMed] [Google Scholar]
  8. Dallai R., T Baldari C., Marchini D., de Filippis T., Rosetto M., Manetti A. G.O. Juvenile hormone regulates the expression of the gene encoding ceratotoxin a, an antibacterial peptide from the female reproductive accessory glands of the medfly Ceratitis capitata. J Insect Physiol. 1997 Nov;43(12):1161–1167. doi: 10.1016/s0022-1910(97)00052-8. [DOI] [PubMed] [Google Scholar]
  9. Diamond G., Jones D. E., Bevins C. L. Airway epithelial cells are the site of expression of a mammalian antimicrobial peptide gene. Proc Natl Acad Sci U S A. 1993 May 15;90(10):4596–4600. doi: 10.1073/pnas.90.10.4596. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Diamond G., Russell J. P., Bevins C. L. Inducible expression of an antibiotic peptide gene in lipopolysaccharide-challenged tracheal epithelial cells. Proc Natl Acad Sci U S A. 1996 May 14;93(10):5156–5160. doi: 10.1073/pnas.93.10.5156. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Dimarcq J. L., Hoffmann D., Meister M., Bulet P., Lanot R., Reichhart J. M., Hoffmann J. A. Characterization and transcriptional profiles of a Drosophila gene encoding an insect defensin. A study in insect immunity. Eur J Biochem. 1994 Apr 1;221(1):201–209. doi: 10.1111/j.1432-1033.1994.tb18730.x. [DOI] [PubMed] [Google Scholar]
  12. Dimopoulos G., Richman A., Müller H. M., Kafatos F. C. Molecular immune responses of the mosquito Anopheles gambiae to bacteria and malaria parasites. Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11508–11513. doi: 10.1073/pnas.94.21.11508. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Driever W., Siegel V., Nüsslein-Volhard C. Autonomous determination of anterior structures in the early Drosophila embryo by the bicoid morphogen. Development. 1990 Aug;109(4):811–820. doi: 10.1242/dev.109.4.811. [DOI] [PubMed] [Google Scholar]
  14. Fearon D. T. Seeking wisdom in innate immunity. Nature. 1997 Jul 24;388(6640):323–324. doi: 10.1038/40967. [DOI] [PubMed] [Google Scholar]
  15. Fehlbaum P., Bulet P., Michaut L., Lagueux M., Broekaert W. F., Hetru C., Hoffmann J. A. Insect immunity. Septic injury of Drosophila induces the synthesis of a potent antifungal peptide with sequence homology to plant antifungal peptides. J Biol Chem. 1994 Dec 30;269(52):33159–33163. [PubMed] [Google Scholar]
  16. Goldman M. J., Anderson G. M., Stolzenberg E. D., Kari U. P., Zasloff M., Wilson J. M. Human beta-defensin-1 is a salt-sensitive antibiotic in lung that is inactivated in cystic fibrosis. Cell. 1997 Feb 21;88(4):553–560. doi: 10.1016/s0092-8674(00)81895-4. [DOI] [PubMed] [Google Scholar]
  17. Heim R., Cubitt A. B., Tsien R. Y. Improved green fluorescence. Nature. 1995 Feb 23;373(6516):663–664. doi: 10.1038/373663b0. [DOI] [PubMed] [Google Scholar]
  18. Hergannan J. A., Rechhart J. V. Drosophila immunity. Trends Cell Biol. 1997 Aug;7(8):309–316. doi: 10.1016/S0962-8924(97)01087-8. [DOI] [PubMed] [Google Scholar]
  19. Higuchi R., Krummel B., Saiki R. K. A general method of in vitro preparation and specific mutagenesis of DNA fragments: study of protein and DNA interactions. Nucleic Acids Res. 1988 Aug 11;16(15):7351–7367. doi: 10.1093/nar/16.15.7351. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Hoffmann J. A., Reichhart J. M., Hetru C. Innate immunity in higher insects. Curr Opin Immunol. 1996 Feb;8(1):8–13. doi: 10.1016/s0952-7915(96)80098-7. [DOI] [PubMed] [Google Scholar]
  21. Ingham P. Signal transduction. Dorsal developments. Nature. 1994 Dec 8;372(6506):500–501. doi: 10.1038/372500a0. [DOI] [PubMed] [Google Scholar]
  22. Janeway C. A., Jr Approaching the asymptote? Evolution and revolution in immunology. Cold Spring Harb Symp Quant Biol. 1989;54(Pt 1):1–13. doi: 10.1101/sqb.1989.054.01.003. [DOI] [PubMed] [Google Scholar]
  23. Kopp E. B., Ghosh S. NF-kappa B and rel proteins in innate immunity. Adv Immunol. 1995;58:1–27. doi: 10.1016/s0065-2776(08)60618-5. [DOI] [PubMed] [Google Scholar]
  24. Kraaijeveld A. R., Godfray H. C. Trade-off between parasitoid resistance and larval competitive ability in Drosophila melanogaster. Nature. 1997 Sep 18;389(6648):278–280. doi: 10.1038/38483. [DOI] [PubMed] [Google Scholar]
  25. Kylsten P., Samakovlis C., Hultmark D. The cecropin locus in Drosophila; a compact gene cluster involved in the response to infection. EMBO J. 1990 Jan;9(1):217–224. doi: 10.1002/j.1460-2075.1990.tb08098.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Lehane M. J., Wu D., Lehane S. M. Midgut-specific immune molecules are produced by the blood-sucking insect Stomoxys calcitrans. Proc Natl Acad Sci U S A. 1997 Oct 14;94(21):11502–11507. doi: 10.1073/pnas.94.21.11502. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Lemaitre B., Nicolas E., Michaut L., Reichhart J. M., Hoffmann J. A. The dorsoventral regulatory gene cassette spätzle/Toll/cactus controls the potent antifungal response in Drosophila adults. Cell. 1996 Sep 20;86(6):973–983. doi: 10.1016/s0092-8674(00)80172-5. [DOI] [PubMed] [Google Scholar]
  28. Levashina E. A., Ohresser S., Bulet P., Reichhart J. M., Hetru C., Hoffmann J. A. Metchnikowin, a novel immune-inducible proline-rich peptide from Drosophila with antibacterial and antifungal properties. Eur J Biochem. 1995 Oct 15;233(2):694–700. doi: 10.1111/j.1432-1033.1995.694_2.x. [DOI] [PubMed] [Google Scholar]
  29. Medzhitov R., Janeway C. A., Jr Innate immunity: impact on the adaptive immune response. Curr Opin Immunol. 1997 Feb;9(1):4–9. doi: 10.1016/s0952-7915(97)80152-5. [DOI] [PubMed] [Google Scholar]
  30. Morisato D., Anderson K. V. Signaling pathways that establish the dorsal-ventral pattern of the Drosophila embryo. Annu Rev Genet. 1995;29:371–399. doi: 10.1146/annurev.ge.29.120195.002103. [DOI] [PubMed] [Google Scholar]
  31. Ouellette A. J., Selsted M. E. Paneth cell defensins: endogenous peptide components of intestinal host defense. FASEB J. 1996 Sep;10(11):1280–1289. doi: 10.1096/fasebj.10.11.8836041. [DOI] [PubMed] [Google Scholar]
  32. Rosetto M., Manetti A. G., Giordano P. C., Marri L., Amons R., Baldari C. T., Marchini D., Dallai R. Molecular characterization of ceratotoxin C, a novel antibacterial female-specific peptide of the ceratotoxin family from the medfly Ceratitis capitata. Eur J Biochem. 1996 Oct 15;241(2):330–337. doi: 10.1111/j.1432-1033.1996.00330.x. [DOI] [PubMed] [Google Scholar]
  33. Russell J. P., Diamond G., Tarver A. P., Scanlin T. F., Bevins C. L. Coordinate induction of two antibiotic genes in tracheal epithelial cells exposed to the inflammatory mediators lipopolysaccharide and tumor necrosis factor alpha. Infect Immun. 1996 May;64(5):1565–1568. doi: 10.1128/iai.64.5.1565-1568.1996. [DOI] [PMC free article] [PubMed] [Google Scholar]
  34. Samakovlis C., Kylsten P., Kimbrell D. A., Engström A., Hultmark D. The andropin gene and its product, a male-specific antibacterial peptide in Drosophila melanogaster. EMBO J. 1991 Jan;10(1):163–169. doi: 10.1002/j.1460-2075.1991.tb07932.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Schonwetter B. S., Stolzenberg E. D., Zasloff M. A. Epithelial antibiotics induced at sites of inflammation. Science. 1995 Mar 17;267(5204):1645–1648. doi: 10.1126/science.7886453. [DOI] [PubMed] [Google Scholar]
  36. Smith J. J., Travis S. M., Greenberg E. P., Welsh M. J. Cystic fibrosis airway epithelia fail to kill bacteria because of abnormal airway surface fluid. Cell. 1996 Apr 19;85(2):229–236. doi: 10.1016/s0092-8674(00)81099-5. [DOI] [PubMed] [Google Scholar]
  37. Steel D. M., Whitehead A. S. The major acute phase reactants: C-reactive protein, serum amyloid P component and serum amyloid A protein. Immunol Today. 1994 Feb;15(2):81–88. doi: 10.1016/0167-5699(94)90138-4. [DOI] [PubMed] [Google Scholar]
  38. Stolzenberg E. D., Anderson G. M., Ackermann M. R., Whitlock R. H., Zasloff M. Epithelial antibiotic induced in states of disease. Proc Natl Acad Sci U S A. 1997 Aug 5;94(16):8686–8690. doi: 10.1073/pnas.94.16.8686. [DOI] [PMC free article] [PubMed] [Google Scholar]
  39. Tryselius Y., Samakovlis C., Kimbrell D. A., Hultmark D. CecC, a cecropin gene expressed during metamorphosis in Drosophila pupae. Eur J Biochem. 1992 Feb 15;204(1):395–399. doi: 10.1111/j.1432-1033.1992.tb16648.x. [DOI] [PubMed] [Google Scholar]
  40. Verma I. M., Stevenson J. K., Schwarz E. M., Van Antwerp D., Miyamoto S. Rel/NF-kappa B/I kappa B family: intimate tales of association and dissociation. Genes Dev. 1995 Nov 15;9(22):2723–2735. doi: 10.1101/gad.9.22.2723. [DOI] [PubMed] [Google Scholar]
  41. Wang S., Hazelrigg T. Implications for bcd mRNA localization from spatial distribution of exu protein in Drosophila oogenesis. Nature. 1994 Jun 2;369(6479):400–403. doi: 10.1038/369400a0. [DOI] [PubMed] [Google Scholar]
  42. Wicker C., Reichhart J. M., Hoffmann D., Hultmark D., Samakovlis C., Hoffmann J. A. Insect immunity. Characterization of a Drosophila cDNA encoding a novel member of the diptericin family of immune peptides. J Biol Chem. 1990 Dec 25;265(36):22493–22498. [PubMed] [Google Scholar]
  43. Zhao C., Wang I., Lehrer R. I. Widespread expression of beta-defensin hBD-1 in human secretory glands and epithelial cells. FEBS Lett. 1996 Nov 4;396(2-3):319–322. doi: 10.1016/0014-5793(96)01123-4. [DOI] [PubMed] [Google Scholar]

Articles from The EMBO Journal are provided here courtesy of Nature Publishing Group

RESOURCES