Advertisement

Fat body and hemocyte contribution to the antimicrobial peptide synthesis in Calliphora vicina R.-D. (Diptera: Calliphoridae) larvae

  • Andrey Yu YakovlevEmail author
  • Alexander P. Nesin
  • Nina P. Simonenko
  • Natalia A. Gordya
  • Dmitriy V. Tulin
  • Anastasia A. Kruglikova
  • Sergey I. Chernysh
Article

Abstract

Antimicrobial peptides accumulated in the hemolymph in response to infection are a key element of insect innate immunity. The involvement of the fat body and hemocytes in the antimicrobial peptide synthesis is widely acknowledged, although release of the peptides present in the hemolymph from the immune cells was not directly verified so far. Here, we studied the presence of antimicrobial peptides in the culture medium of fat body cells and hemocytes isolated from the blue blowfly Calliphora vicina using complex of liquid chromatography, mass spectrometry, and antimicrobial activity assays. Both fat body and hemocytes are shown to synthesize and release to culture medium defensin, cecropin, diptericins, and proline-rich peptides. The spectra of peptide antibiotics released by the fat body and hemocytes partially overlap. Thus, the results suggest that insect fat body and blood cells are capable of releasing mature antimicrobial peptides to the hemolymph. It is notable that the data obtained demonstrate dramatic difference in the functioning of insect antimicrobial peptides and their mammalian counterparts localized into blood cells’ phagosomes where they exert their antibacterial activity.

Keywords

Insect immunity Antimicrobial peptides Cell cultures Fat body Hemocytes 

Notes

Acknowledgments

The work was founded by the Russian Science Foundation (grant no. 16-14-00048). The research was supported by the St. Petersburg State University’s research resource centers «Molecular and cell technologies» and «Сenter for chemical analysis and materials research».

References

  1. Bartholomay LC, Cho WL, Rocheleau TA, et al. (2004) Description of the transcriptomes of immune response-activated hemocytes from the mosquito vectors Aedes aegypti and Armigeres subalbatus. Infect Immunol 72:4114–4126CrossRefGoogle Scholar
  2. Boman HG, Faye I, Pye A, Rasmuson T (1978) The inducible immunity system of giant silk moths. Comp Pathobiol 4:145–163Google Scholar
  3. Boulanger N, Brun R, Ehret-Sabatier L, Kunz C, Bulet P (2002) Immunopeptides in the defense reactions of Glossina morsitans to bacterial and Trypanosoma brucei brucei infections. Insect Biochem Mol Biol 32(4):369–375CrossRefPubMedGoogle Scholar
  4. Bulet P, Cociancich S, Dimarcq JL, Lambert J, Reichhart JM, Hoffmann D, et al. (1991) Insect immunity. Isolation from a coleopteran insect of a novel inducible antibacterial peptide and of new members of the insect defensin family. J Biol Chem 36:24520–24525Google Scholar
  5. Bulet P, Stöcklin R (2005) Insect antimicrobial peptides: structures, properties and gene regulation. Protein Pept Lett 12:3–11CrossRefPubMedGoogle Scholar
  6. Caicedo AP, Fernandez ST, Arias J, Hernandez L (2014) Verification of pharmaceutical power of generic meropenem vs innovator trough assessment by microbial potency test. Afr J Pharm Pharmacol 8(37):909–916Google Scholar
  7. Carlson JG, Hollaender A, Gaulden ME (1947) Ultraviolet radiation as a means of sterilizing tissue culture materials. Science 105:187–188CrossRefPubMedGoogle Scholar
  8. Cerovský V, Zdárek J, Fucík V, et al. (2010) Lucifensin, the long-sought antimicrobial factor of medicinal maggots of the blowfly Lucilia sericata. Cell Mol Life Sci 67:455–466CrossRefPubMedGoogle Scholar
  9. Chernysh SI, Gordja NA (2011) The immune system of maggots of the blow fly (Calliphora vicina) as a source of medicinal drugs. J Evol Biochem Physiol 47:524–533CrossRefGoogle Scholar
  10. Chernysh SI, Gordja NA, Simonenko NP (2000) Diapause and immune response: induction of antimicrobial peptides synthesis in the blowfly, Calliphora vicina R.-D. (Diptera, Calliphoridae). Entomol Sci 3:139–144Google Scholar
  11. Chernysh S, Gordya N, Suborova T (2015) Insect antimicrobial peptide complexes prevent resistance development in bacteria. PLoS One 10:e0130788CrossRefPubMedPubMedCentralGoogle Scholar
  12. Christophides GK, Zdobnov E, Barillas-Mury C, et al. (2002) Immunity-related genes and gene families in Anopheles gambiae. Science 298:159–165CrossRefPubMedGoogle Scholar
  13. Cociancich S, Ghazi A, Hetru C, Hoffmann JA, Letellier L (1993) Insect defensin, an inducible antibacterial peptide, forms voltage-dependent channels in Micrococcus luteus. J Biol Chem 268(26):19239–19245PubMedGoogle Scholar
  14. Dimarcq JL, Imler JL, Lanot R, et al. (1997) Treatment of l(2)mbn drosophila tumorous blood cells with the steroid hormone ecdysone amplifies the inducibility of antimicrobial peptide gene expression. Insect Biochem Mol Biol 27:877–886CrossRefPubMedGoogle Scholar
  15. Dimarcq JL, Keppi E, Dunbar B, et al. (1988) Purification and characterization of a family of novel inducible antibacterial proteins from immunized larvae of the dipteran Phormia terranovae and complete amino-acid sequence of the predominant member, diptericin a. Eur J Biochem 171:17–22CrossRefPubMedGoogle Scholar
  16. Faye I, Wyatt GR (1980) The synthesis of antibacterial proteins in isolated fat body from cecropia silkmoth pupae. Experientia 36:1325–1326CrossRefPubMedGoogle Scholar
  17. Ganz T, Selsted ME, Szklarek D, Harwig SS, Daher K, Bainton DF, Lehrer RI (1985) Defensins. Natural peptide antibiotics of human neutrophils. J Clin Invest 76:1427–1435CrossRefPubMedPubMedCentralGoogle Scholar
  18. Hoffmann JA, Reichhart L-M (1997) Drosophila immunity. Trends Cell Biol 7:309–316CrossRefGoogle Scholar
  19. Hultmark D (1993) Immune reactions in drosophila and other insects: a model for innate immunity. J Tr Genet 9:178–183CrossRefGoogle Scholar
  20. Irving P, Ubeda JM, Doucet D, et al. (2005) New insights into drosophila larval haemocyte functions through genome-wide analysis. Cell Microbiol 7:335–350CrossRefPubMedGoogle Scholar
  21. Johansson KC, Metzendorf C, Soderhall K (2005) Microarray analysis of immune challenged drosophila hemocytes. Exp Cell Res 305:145–155CrossRefPubMedGoogle Scholar
  22. Kind TV (2003) Hemocytes of the blowfly Calliphora vicina and their dynamics during larval development and metamorphosis. Tsitologiya 45:14–25 [in Russian]Google Scholar
  23. Kind TV (2007) Hemocytes demonstrating different types of protective response in the ontogeny of three species of blowfly genus Calliphora. BiNII St. Petersburg State University Proceedings 53:306–335 [in Russian]Google Scholar
  24. Kind TV (2012) Functional morphology of blowfly Calliphora vicina hemocytes. Tsitologiya 54:806–822 [in Russian]Google Scholar
  25. Kokryakov VN (1999) Biology of antibiotics from animal sources. Nauka, St. Petersburg [in Russian]Google Scholar
  26. Kruglikova AA, Chernysh SI (2011) Antimicrobial compounds from the excretions of surgical maggots, Lucilia sericata (Meigen) (Diptera, Calliphoridae). Entomol Rev 91:813–819CrossRefGoogle Scholar
  27. Lambert J, Keppi E, Dimarcq J-L (1989) Insect immunity: isolation from immune blood of the dipteran Phormia terranovae of two insect antibacterial peptides with sequence homology to rabbit lung macrophage bacteri_cidal peptides. Proc Nat Acad Sci USA 86:262–266CrossRefPubMedPubMedCentralGoogle Scholar
  28. Mitsuhashi J, Maramorosch K (1964) Leafhopper tissue culture: embryonic, nymphal and imaginal tissues from aseptic insects. Contrib Boyce Thompson Inst 22:435–460Google Scholar
  29. Pitkin DH, Sheikh W, Nadler HL (1997) Comparative in vitro activity of meropenem versus other extended-spectrum antimicrobials against randomly chosen and selected resistant clinical isolates tested in 26 north American centers. Clin Infect Dis 24(2):S238–S248CrossRefPubMedGoogle Scholar
  30. Tang T, Li X, Yang X, et al. (2014) Transcriptional response of Musca domestica larvae to bacterial infection. PLoS One. doi: 10.1371/Journal.Pone.0104867 Google Scholar
  31. Trenszec T, Faye I (1988) Synthesis of immune proteins in primary cultures of fat body from Hyalophora cecropia. J Ins Bio chem 18:299–312Google Scholar
  32. Tulin DV, Chaga OI (2003) Hemocytes of Calliphora vicina larvae. I Histol Anal Tsitol 45:976–985 [in Russian]Google Scholar
  33. Yakovlev AY (2011) Induction of antimicrobial peptide synthesis by the fat body cells of maggots of Calliphora vicina R.-D. (Diptera, Calliphoridae). J Evol Biochem Physiol 47:543–551CrossRefGoogle Scholar
  34. Yamada K, Natori S (1993) Purification, sequence and antibacterial activity of two novel sapecin homologues from Sarcophaga embryonic cells: similarity of sapecin B to charybdotoxin. Biochem J 291:275–279CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© The Society for In Vitro Biology 2016

Authors and Affiliations

  • Andrey Yu Yakovlev
    • 1
    Email author
  • Alexander P. Nesin
    • 1
  • Nina P. Simonenko
    • 1
  • Natalia A. Gordya
    • 1
  • Dmitriy V. Tulin
    • 1
  • Anastasia A. Kruglikova
    • 1
  • Sergey I. Chernysh
    • 1
  1. 1.Laboratory of Insect Biopharmacology and Immunology, Department of EntomologySaint-Petersburg State UniversitySaint-PetersburgRussia

Personalised recommendations