Insect Hemolymph Immune Complexes

  • Kevin D. ClarkEmail author
Part of the Subcellular Biochemistry book series (SCBI, volume 94)


Insects possess powerful immune systems that have evolved to defend against wounding and environmental pathogens such as bacteria, fungi, protozoans, and parasitoids. This surprising sophistication is accomplished through the activation of multiple immune pathways comprised of a large array of components, many of which have been identified and studied in detail using both genetic manipulations and traditional biochemical techniques. Recent advances indicate that certain pathways activate arrays of proteins that interact to form large functional complexes. Here we discuss three examples from multiple insects that exemplify such processes, including pathogen recognition, melanization, and coagulation. The functionality of each depends on integrating recognition with the recruitment of immune effectors capable of healing wounds and destroying pathogens. In both melanization and coagulation, protein interactions also appear to be essential for enzymatic activities tied to the formation of melanin and for the recruitment of hemocytes. The importance of these immune complexes is highlighted by the evolution of mechanisms in pathogens to disrupt their formation, an example of which is provided. While technically difficult to study, and not always readily amenable to dissection through genetics, modern mass spectrometry has become an indispensable tool in the study of these higher-order protein interactions. The formation of immune complexes should be viewed as an essential and emerging frontier in the study of insect immunity.


Insect immunity Pattern recognition receptors Pathogen-associated molecular patterns C3 proteins TEP1 convertase Thioester proteins Leucine repeat proteins Clip-domain serine proteases Serine protease homologs Tyrosinases Melanization Phenoloxidase Phenoloxidase cascade inhibitors Hemocyanins Coagulation Coagulogen Clotting proteins Clot associated proteins Hemocytes Lipopolysaccharides Transglutaminase Plasmatocyte spreading peptide 







Melanization complex


Immune complex


Hemolymph proteases


Phenoloxidase-activating protease


Serine protease homolog






An arthropod-specific class of SPs and SPHs








Green fluorescent protein




Plasmatocyte spreading peptide


Reactive oxygen species


Antimicrobial peptide


Sodium dodecyl sulfate-polyacrylamide gel electrophoresis


Pathogen-associated molecular pattern




Substrate-free plasma


Pattern recognition receptor


N-terminal catalytic domain


C-terminal repeat domain


  1. Agianian B, Lesch C, Loseva O, Dushay MS (2007) Preliminary characterization of hemolymph coagulation in Anopheles gambiae larvae. Dev Comp Immunol 31(9):879–888. Scholar
  2. An C, Budd A, Kanost M, Michel K (2011) Characterization of a regulatory unit that controls melanization and affects longevity of mosquitoes. Cell Mol Life Sci 68(11):1929–1939. Scholar
  3. Asgari S (2006) Venom proteins from polydnavirus-producing endoparasitoids: their role in host-parasite interactions. Arch Insect Biochem Physiol 61(3):146–156CrossRefGoogle Scholar
  4. Aspán A, Huang TS, Cerenius L, Söderhäll K (1995) cDNA cloning of prophenoloxidase from the freshwater crayfish Pacifastacus leniusculus and its activation. Proc Natl Acad Sci USA 92(4):939–943CrossRefGoogle Scholar
  5. Barwig B (1985) Isolation and characterization of plasma coagulogen (PC) of the cockroach Leucophaea maderae (Blattaria). J Comp Physiol B 155(2):135–143. Scholar
  6. Baxter RHG, Steinert S, Chelliah Y, Volohonsky G, Levashina EA, Deisenhofer J (2010) A heterodimeric complex of the LRR proteins LRIM1 and APL1C regulates complement-like immunity in Anopheles gambiae. Proc Natl Acad Sci USA 107(39):16817. Scholar
  7. Beck MH, Strand MR (2007) A novel polydnavirus protein inhibits the insect prophenoloxidase activation pathway. Proc Natl Acad Sci USA 104(49):19267–19272. Scholar
  8. Beckage NE, Kanost MR (1993) Effects of parasitism by the braconid wasp Cotesia congregata on host hemolymph proteins of the tobacco hornworm. Manduca sexta. Insect Biochem Mol Biol 23(5):643–653. Scholar
  9. Bella J, Hindle KL, McEwan PA, Lovell SC (2008) The leucine-rich repeat structure. Cell Mol Life Sci 65(15):2307–2333. Scholar
  10. Berisha A, Mukherjee K, Vilcinskas A, Spengler B, Römpp A (2013) High-resolution mass spectrometry driven discovery of peptidic danger signals in insect immunity. PLoS ONE 8(11):e80406. Scholar
  11. Bidla G, Dushay MS, Theopold U (2007) Crystal cell rupture after injury in Drosophila requires the JNK pathway, small GTPases and the TNF homolog Eiger. J Cell Sci 120(7):1209–1215. Scholar
  12. Bidla G, Hauling T, Dushay MS, Theopold U (2009) Activation of Insect phenoloxidase after injury: endogenous versus foreign elicitors. J Innate Immun 1(4):301–308CrossRefGoogle Scholar
  13. Bidla G, Lindgren M, Theopold U, Dushay MS (2005) Hemolymph coagulation and phenoloxidase in Drosophila larvae. Dev Comp Immunol 29(8):669–679. Scholar
  14. Blandin S, Levashina EA (2004) Thioester-containing proteins and insect immunity. Mol Immunol 40(12):903–908CrossRefGoogle Scholar
  15. Blandin S, Shiao S-H, Moita LF, Janse CJ, Waters AP, Kafatos FC, Levashina EA (2004) Complement-like protein TEP1 Is a determinant of vectorial capacity in the malaria vector Anopheles gambiae. Cell 116(5):661–670. Scholar
  16. Bohn H, Barwig B (1984) Hemolymph clotting in the cockroach Leucophaea maderae (Blattaria). J Comp Physiol B 154(5):457–467. Scholar
  17. Brack A, Hellmann N, Decker H (2008) Kinetic properties of hexameric tyrosinase from the crustacean palinurus elephas. Photochem Photobiol 84(3):692–699. Scholar
  18. Burmester T (2001) Molecular evolution of the arthropod hemocyanin superfamily. Mol Biol Evol 18(2):184–195CrossRefGoogle Scholar
  19. Cerenius L, Söderhäll K (2010) Coagulation in invertebrates. J Innate Immun 3(1):3–8CrossRefGoogle Scholar
  20. Chang H-J, Dhanasingh I, Gou X, Rice AM, Dushay MS (2012) Loss of Hemolectin reduces the survival of Drosophila larvae after wounding. Dev Comp Immunol 36(2):274–278. Scholar
  21. Clark KD (2015) Altered tyrosine metabolism and melanization complex formation underlie the developmental regulation of melanization in Manduca sexta. Insect Biochem Mol Biol 58:66–75. Scholar
  22. Clark KD, Kim Y, Strand MR (2005) Plasmatocyte sensitivity to plasmatocyte spreading peptide (PSP) fluctuates with the larval molting cycle. J Insect Physiol 51(5):587–596. Scholar
  23. Clark KD, Lu Z, Strand MR (2010) Regulation of melanization by glutathione in the moth Pseudoplusia includens. Insect Biochem Mol Biol 40(6):460–467CrossRefGoogle Scholar
  24. Clark KD, Pech LL, Strand MR (1997) Isolation and identification of a plasmatocyte-spreading peptide from the hemolymph of the lepidopteran insect pseudoplusia includens. J Biol Chem 272(37):23440–23447. Scholar
  25. Clark KD, Strand MR (2013) Hemolymph melanization in the silkmoth bombyx mori involves formation of a high molecular mass complex that metabolizes tyrosine. J Biol Chem 288(20):14476–14487. Scholar
  26. Decker H, Schweikardt T, Nillius D, Salzbrunn U, Jaenicke E, Tuczek F (2007) Similar enzyme activation and catalysis in hemocyanins and tyrosinases. Gene 398(1):183–191. Scholar
  27. Delvaeye M, Conway EM (2009) Coagulation and innate immune responses: can we view them separately? Blood 114(12):2367–2374. Scholar
  28. Dodds AW, Law SKA (1998) The phylogeny and evolution of the thioester bond-containing proteins C3, C4 and and alpha-2-macroglobulin. Immunol Rev 166(1):15–26CrossRefGoogle Scholar
  29. Dong Y, Aguilar R, Xi Z, Warr E, Mongin E, Dimopoulos G (2006) Anopheles gambiae immune responses to human and rodent plasmodium parasite species. PLoS Pathog 2(6):e52. Scholar
  30. Dudzic JP, Kondo S, Ueda R, Bergman CM, Lemaitre B (2015) Drosophila innate immunity: regional and functional specialization of prophenoloxidases. BMC Biol 13(1):81. Scholar
  31. Dziedziech A, Schmid M, Arefin B, Kienzle T, Krautz R, Theopold U (2019) Data on Drosophila clots and hemocyte morphologies using GFP-tagged secretory proteins: Prophenoloxidase and transglutaminase. Data in Brief 25:104229. Scholar
  32. Engström Y, Loseva O, Theopold U (2004) Proteomics of the drosophila immune response. Trends Biotechnol 22(11):600–605. Scholar
  33. Felföldi G, Eleftherianos I, ffrench-Constant RH, Venekei I (2011) A Serine Proteinase Homologue, SPH-3, Plays a Central Role in Insect Immunity. J Immunol 186 (8):4828–4834.
  34. Fraiture M, Baxter RHG, Steinert S, Chelliah Y, Frolet C, Quispe-Tintaya W, Hoffmann JA, Blandin SA, Levashina EA (2009) Two mosquito LRR proteins function as complement control factors in the TEP1-mediated killing of plasmodium. Cell Host Microbe 5(3):273–284. Scholar
  35. Frolet C, Thoma M, Blandin S, Hoffmann JA, Levashina EA (2006) Boosting NF-kB-dependent basal immunity of Anopheles gambiae aborts development of plasmodium berghei. Immunity 25(4):677–685. Scholar
  36. Goto A, Kadowaki T, Kitagawa Y (2003) Drosophila hemolectin gene is expressed in embryonic and larval hemocytes and its knock down causes bleeding defects. Dev Biol 264(2):582–591. Scholar
  37. Goto A, Kumagai T, Kumagai C, Hirose J, Narita H, Mori H, Kadowaki T, Beck K, Kitagawa Y (2001) A drosophila haemocyte-specific protein, hemolectin, similar to human von willebrand factor. Biochem J 359(1):99–108. Scholar
  38. Gupta S, Wang Y, Jiang H (2005) Manduca sexta prophenoloxidase (proPO) activation requires proPO-activating proteinase (PAP) and serine proteinase homologs (SPHs) simultaneously. Insect Biochem Mol Biol 35(3):241–248CrossRefGoogle Scholar
  39. Hayakawa Y (1994) Cellular immunosuppressive protein in the plasma of parasitized insect larvae. J Biol Chem 269(20):14536–14540PubMedGoogle Scholar
  40. He Y, Cao X, Zhang S, Rogers J, Hartson S, Jiang H (2016) Changes in the plasma proteome of manduca sexta larvae in relation to the transcriptome variations after an immune challenge: evidence for high molecular weight immune complex formation. Mol Cell Proteomics: MCP 15(4):1176–1187. Scholar
  41. He Y, Wang Y, Yang F, Jiang H (2017) Manduca sexta hemolymph protease-1, activated by an unconventional non-proteolytic mechanism, mediates immune responses. Insect Biochem Mol Biol 84:23–31. Scholar
  42. Heck T, Faccio G, Richter M, Thöny-Meyer L (2013) Enzyme-catalyzed protein crosslinking. Appl Microbiol Biotechnol 97(2):461–475. Scholar
  43. Hughes AL (1999) Evolution of the arthropod prophenoloxidase/hexamerin protein family. Immunogenetics 49(2):106–114. Scholar
  44. Iijima M, Hashimoto T, Matsuda Y, Nagai T, Yamano Y, Ichi T, Osaki T, Kawabata S-I (2005) Comprehensive sequence analysis of horseshoe crab cuticular proteins and their involvement in transglutaminase-dependent cross-linking. FEBS J 272(18):4774–4786. Scholar
  45. Ito S (2003) A chemist’s view of melanogenesis. Pigment Cell Res 16(3):230–236. Scholar
  46. Ito S, Palumbo A, Prota G (1985) Tyrosinase-cytalyzed conjugation of dopa with glutathione. Cell Mol Life Sci 41(7):960–961. Scholar
  47. Iwanaga S (2002) The molecular basis of innate immunity in the horseshoe crab. Curr Opin Immunol 14(1):87–95. Scholar
  48. Iwanaga S, Lee BL (2005) Recent advances in the innate immunity of invertebrate animals. J Biochem Mol Biol 38(2):128–150PubMedGoogle Scholar
  49. Jaenicke E, Decker H (2003) Tyrosinases from crustaceans form hexamers. Biochem J 371:515–523CrossRefGoogle Scholar
  50. Jiang H, Vilcinskas A, Kanost MR (2010) Immunity in lepidopteran insects. In: Söderhäll K (ed) Invertebrate Immunity, vol 708. Advances in Experimental Medicine and Biology. Springer US, pp 181–204.
  51. Jouni ZE, Takada N, Gazard J, Maekawa H, Wells MA, Tsuchida K (2003) Transfer of cholesterol and diacylglycerol from lipophorin to Bombyx mori ovarioles in vitro: role of the lipid transfer particle. Insect Biochem Mol Biol 33(2):145–153. Scholar
  52. Kamareddine L, Nakhleh J, Osta MA (2016) Functional interaction between Apolipophorins and complement regulate the mosquito immune response to systemic infections. J Innate Immun 8(3):314–326. Scholar
  53. Kamimura M, Nakahara Y, Kanamori Y, Tsuzuki S, Hayakawa Y, Kiuchi M (2001) Molecular cloning of silkworm paralytic peptide and its developmental regulation. Biochem Biophys Res Commun 286(1):67–73CrossRefGoogle Scholar
  54. Kan H, Kim C-H, Kwon H-M, Park J-W, Roh K-B, Lee H, Park B-J, Zhang R, Zhang J, Söderhäll K, Ha N-C, Lee BL (2008) Molecular control of phenoloxidase-induced melanin synthesis in an insect. J Biol Chem 283(37):25316–25323. Scholar
  55. Kanost MR, Jiang H (2015) Clip-domain serine proteases as immune factors in insect hemolymph. Curr Opin Insect Sci 11:47–55. Scholar
  56. Karlsson C, Korayem AM, Scherfer C, Loseva O, Dushay MS, Theopold U (2004) Proteomic analysis of the drosophila larval hemolymph clot. J Biol Chem 279(50):52033–52041. Scholar
  57. Kato Y, Motoi Y, Taniai K, Kadono-Okuda K, Yamamoto M, Higashino Y, Shimabukuro M, Chowdhury S, Xu J, Sugiyama M, Hiramatsu M, Yamakawa M (1994) Lipopolysaccharide-lipophorin complex formation in insect hemolymph: a common pathway of lipopolysaccharide detoxification both in insects and in mammals. Insect Biochem Mol Biol 24(6):547–555. Scholar
  58. Kim MS, Baek MJ, Lee MH, Park JW, Lee SY, Söderhäll K, Lee BL (2002) A new easter-type serine protease cleaves a masquerade-like protein during prophenoloxidase activation in holotrichia diomphalia larvae. J Biol Chem 277(42):39999–40004. Scholar
  59. Kim-Jo C, Gatti J-L, Poirié M (2019) Drosophila cellular immunity against parasitoid wasps: a complex and time-dependent process. Front Physiol 10(603).
  60. Kotani E, Yamakawa M, Iwamoto S-i, Tashiro M, Mori H, Sumida M, Matsubara F, Taniai K, Kadono-Okuda K, Kato Y, Mori H (1995) Cloning and expression of the gene of hemocytin, an insect humoral lectin which is homologous with the mammalian von Willebrand factor. Biochim Biophys Acta 1260(3):245–258. Scholar
  61. Krautz R, Arefin B, Theopold U (2014) Damage signals in the insect immune response. Front Plant Sci 5(342).
  62. Kwon TH, Kim MS, Choi HW, Joo CH, Cho MY, Lee BL (2000) A masquerade-like serine proteinase homologue is necessary for phenoloxidase activity in the coleopteran insect, Holotrichia diomphalia larvae. Eur J Biochem 267(20):6188–6196CrossRefGoogle Scholar
  63. Lagueux M, Perrodou E, Levashina EA, Capovilla M, Hoffmann JA (2000) Constitutive expression of a complement-like protein in Toll and JAK gain-of-function mutants of Drosophila. Proc Natl Acad Sci USA 97(21):11427. Scholar
  64. Lee SY, Kwon TH, Hyun JH, Choi JS, Kawabata S-I, Iwanaga S, Lee BL (1998) In vitro activation of pro-phenol-oxidase by two kinds of pro-phenol-oxidase-activating factors isolated from hemolymph of coleopteran, Holotrichia diomphalia larvae. Eur J Biochem 254(1):50–57. Scholar
  65. Lesch C, Goto A, Lindgren M, Bidla G, Dushay MS, Theopold U (2007) A role for Hemolectin in coagulation and immunity in Drosophila melanogaster. Dev Comp Immunol 31(12):1255–1263. Scholar
  66. Levashina EA, Moita LF, Blandin S, Vriend G, Lagueux M, Kafatos FC (2001) Conserved Role of a complement-like protein in Phagocytosis revealed by dsRNA knockout in cultured cells of the mosquito, Anopheles gambiae. Cell 104(5):709–718. Scholar
  67. Levy F, Bulet P, Ehret-Sabatier L (2004) Proteomic analysis of the systemic immune response of Drosophila. Mol Cell Proteomics 3(2):156. Scholar
  68. Li D, Scherfer C, Korayem AM, Zhao Z, Schmidt O, Theopold U (2002) Insect hemolymph clotting: evidence for interaction between the coagulation system and the prophenoloxidase activating cascade. Insect Biochem Mol Biol 32(8):919–928. Scholar
  69. Li X, Ma M, Liu F, Chen Y, Lu A, Ling Q-Z, Li J, Beerntsen BT, Yu X-Q, Liu C, Ling E (2012) Properties of Drosophila melanogaster prophenoloxidases expressed in Escherichia coli. Dev Comp Immunol 36(4):648–656. Scholar
  70. Lin C-Y, Hu K-Y, Ho S-H, Song Y-L (2006) Cloning and characterization of a shrimp clip domain serine protease homolog (c-SPH) as a cell adhesion molecule. Dev Comp Immunol 30(12):1132–1144. Scholar
  71. Lindgren M, Riazi R, Lesch C, Wilhelmsson C, Theopold U, Dushay MS (2008) Fondue and transglutaminase in the Drosophila larval clot. J Insect Physiol 54(3):586–592. Scholar
  72. Lu Z, Beck MH, Strand MR (2010) Egf1.5 is a second phenoloxidase cascade inhibitor encoded by Microplitis demolitor bracovirus. Insect Biochem Mol Biol 40(7):497–505.
  73. Lu Z, Beck MH, Wang Y, Jiang H, Strand MR (2008a) The viral protein Egf1.0 Is a dual activity inhibitor of prophenoloxidase-activating proteinases 1 and 3 from Manduca sexta. J Biol Chem 283(31):21325–21333.
  74. Lu Z, Jiang H (2008b) Expression of Manduca sexta serine proteinase homolog precursors in insect cells and their proteolytic activation. Insect Biochem Mol Biol 38(1):89–98Google Scholar
  75. Matheis G, Belitz H-D (1979) Multiple forms of soluble monophenol, dihydroxyphenylalanine: oxygen oxidoreductase (EC from potato tubers (solanum tuberosum). Z Lebensmittelunters Forsch A 169(3):165–169.
  76. Matsumoto H, Tsuzuki S, Date-Ito A, Ohnishi A, Hayakawa Y (2012) Characteristics common to a cytokine family spanning five orders of insects. Insect Biochem Mol Biol 42(6):446–454. Scholar
  77. Moita LF, Wang-Sattler R, Michel K, Zimmermann T, Blandin S, Levashina EA, Kafatos FC (2005) In vivo identification of novel regulators and conserved pathways of Phagocytosis in A. gambiae. Immunity 23(1):65–73.
  78. Morais Guedes Sd, Vitorino R, Domingues R, Tomer K, Correia AJF, Amado F, Domingues P (2005) Proteomics of immune-challenged drosophila melanogaster larvae hemolymph. Biochem Biophys Res Commun 328(1):106–115. Scholar
  79. Mori H, Iwamoto S-i, Kotani E, Sumida M, Matsumoto T, Matsubara F (1992) Isolation of cDNA clones coding for humoral lectin of silkworm, Bombyx mori, larvae. J Invertebr Pathol 59(1):40–45. Scholar
  80. Muszbek L, Bereczky Z, Bagoly Z, Komáromi I, Katona É (2011) Factor XIII: a coagulation factor with multiple plasmatic and cellular functions. Physiol Rev 91(3):931–972. Scholar
  81. Nagai T, Kawabata S-i (2000) A link between blood coagulation and prophenol oxidase activation in arthropod host defense. J Biol Chem 275(38):29264–29267. Scholar
  82. Nakhleh J, Christophides GK, Osta MA (2017) The serine protease homolog CLIPA14 modulates the intensity of the immune response in the mosquito Anopheles gambiae. J Biol Chem 292(44):18217–18226. Scholar
  83. Nappi A, Poirié M, Carton Y (2009) Chapter 4 The role of melanization and cytotoxic by-products in the cellular immune responses of Drosophila against parasitic wasps. In: Genevieve P (ed) Advances in parasitology, vol 70. Academic Press, pp 99–121Google Scholar
  84. Oliveira GdA, Lieberman J, Barillas-Mury C (2012) Epithelial nitration by a peroxidase/NOX5 system mediates mosquito antiplasmodial immunity. Science (New York, NY) 335(6070):856–859. Scholar
  85. Orlow SJ, Zhou B-K, Chakraborty AK, Drucker M, Pifko-Hirst S, Pawelek JM (1994) High-molecular-weight forms of Tyrosinase and the Tyrosinase-related proteins: evidence for a melanogenic complex. J Investig Dermatol 103(2):196–201CrossRefGoogle Scholar
  86. Osta MA, Christophides GK, Kafatos FC (2004) Effects of mosquito genes on plasmodium development. Science 303(5666):2030–2032. Scholar
  87. Paskewitz SM, Andreev O, Shi L (2006) Gene silencing of serine proteases affects melanization of Sephadex beads in Anopheles gambiae. Insect Biochem Mol Biol 36(9):701–711. Scholar
  88. Pedersen LC, Yee VC, Bishop PD, Trong IL, Teller DC, Stenkamp RE (1994) Transglutaminase factor XIII uses proteinase-like catalytic triad to crosslink macromolecules. Protein Sci 3(7):1131–1135. Scholar
  89. Peñalver MJ, Fenoll LG, Rodríguez-López JN, García-Ruiz PA, García-Molina F, Varón R, García-Cánovas F, Tudela J (2005) Reaction mechanism to explain the high kinetic autoactivation of tyrosinase. J Mol Catal B 33(1):35–42. Scholar
  90. Phillips DR, Clark KD (2017) Bombyx mori and Aedes aegypti form multi-functional immune complexes that integrate pattern recognition, melanization, coagulants, and hemocyte recruitment. PLoS ONE 12(2):e0171447. Scholar
  91. Piao S, Song Y-L, Kim JH, Park SY, Park JW, Lee BL, Oh B-H, Ha N-C (2005) Crystal structure of a clip-domain serine protease and functional roles of the clip domains. EMBO J 24(24):4404–4414CrossRefGoogle Scholar
  92. Pietrzyk AJ, Bujacz A, Mueller-Dieckmann J, Łochynska M, Jaskolski M, Bujacz G (2013) Crystallographic identification of an unexpected protein complex in silkworm haemolymph. Acta Crystallogr Sect D: Biol Crystallogr 69(12):2353–2364. Scholar
  93. Povelones M, Bhagavatula L, Yassine H, Tan LA, Upton LM, Osta MA, Christophides GK (2013) The CLIP-domain serine protease homolog SPCLIP1 regulates complement recruitment to microbial surfaces in the malaria mosquito Anopheles gambiae. PLoS Path 9(9):e1003623. Scholar
  94. Povelones M, Upton LM, Sala KA, Christophides GK (2011) Structure-function analysis of the Anopheles gambiae LRIM1/APL1C complex and its interaction with complement C3-Like protein TEP1. PLoS Path 7(4):e1002023. Scholar
  95. Povelones M, Waterhouse RM, Kafatos FC, Christophides GK (2009) Leucine-rich repeat protein complex activates mosquito complement in defense against plasmodium parasites. Science 324(5924):258–261. Scholar
  96. Rahman MM, Ma G, Roberts HLS, Schmidt O (2006) Cell-free immune reactions in insects. J Insect Physiol 52(7):754–762. Scholar
  97. Rebsamen M, Kandasamy RK, Superti-Furga G (2013) Protein interaction networks in innate immunity. Trends Immunol 34(12):610–619. Scholar
  98. Ricklin D, Reis ES, Mastellos DC, Gros P, Lambris JD (2016) Complement component C3—The “Swiss Army Knife” of innate immunity and host defense. Immunol Rev 274(1):33–58. Scholar
  99. Riehle MM, Markianos K, Niaré O, Xu J, Li J, Touré AM, Podiougou B, Oduol F, Diawara S, Diallo M, Coulibaly B, Ouatara A, Kruglyak L, Traoré SF, Vernick KD (2006) Natural malaria infection in Anopheles gambiae is regulated by a single genomic control region. Science 312(5773):577. Scholar
  100. Riehle MM, Xu J, Lazzaro BP, Rottschaefer SM, Coulibaly B, Sacko M, Niare O, Morlais I, Traore SF, Vernick KD (2008) Anopheles gambiae APL1 Is a family of variable LRR proteins required for Rel1-mediated protection from the malaria parasite. Plasmodium berghei. PLOS ONE 3(11):e3672. Scholar
  101. Sakamoto M, Ohta M, Suzuki A, Takase H, Yoshizawa Y, Kitami M, Sato R (2011) Localization of the serine protease homolog BmSPH-1 in nodules of E. coli-injected Bombyx mori larvae and functional analysis of its role in nodule melanization. Dev Comp Immunol 35(5):611–619.
  102. Scherfer C, Karlsson C, Loseva O, Bidla G, Goto A, Havemann J, Dushay MS, Theopold U (2004) Isolation and characterization of hemolymph clotting factors in drosophila melanogaster by a pullout method. Curr Biol 14(7):625–629. Scholar
  103. Schmid MR, Dziedziech A, Arefin B, Kienzle T, Wang Z, Akhter M, Berka J, Theopold U (2019) Insect hemolymph coagulation: kinetics of classically and non-classically secreted clotting factors. Insect Biochem Mol Biol 109:63–71. Scholar
  104. Schmidt O, Söderhäll K, Theopold U, Faye I (2010) Role of Adhesion in arthropod immune recognition. Annu Rev Entomol 55(1):485–504. Scholar
  105. Schnitger AKD, Kafatos FC, Osta MA (2007) The melanization reaction is not required for survival of Anopheles gambiae mosquitoes after bacterial infections. J Biol Chem 282(30):21884–21888. Scholar
  106. Sendovski M, Kanteev M, Ben-Yosef VS, Adir N, Fishman A (2011) First structures of an active bacterial tyrosinase reveal copper plasticity. J Mol Biol 405(1):227–237. Scholar
  107. Seo S-Y, Sharma VK, Sharma N (2003) Mushroom Tyrosinase: recent prospects. J Agric Food Chem 51(10):2837–2853. Scholar
  108. Shapiro JP, Keim PS, Law JH (1984) Structural studies on lipophorin, an insect lipoprotein. J Biol Chem 259(6):3680–3685PubMedGoogle Scholar
  109. Shelby KS, Webb BA (1997) Polydnavirus infection inhibits translation of specific growth-associated host proteins. Insect Biochem Mol Biol 27(3):263–270. Scholar
  110. Shibata T, Hadano J, Kawasaki D, Dong X, Kawabata S-i (2017a) Drosophila TG-A transglutaminase is secreted via an unconventional Golgi-independent mechanism involving exosomes and two types of fatty acylations. J Biol Chem 292(25):10723–10734. Scholar
  111. Shokal U, Eleftherianos I (2017b) Thioester-containing protein-4 regulates the drosophila immune signaling and function against the pathogen photorhabdus. J Innate Immun 9(1):83–93.
  112. Shokal U, Kopydlowski H, Eleftherianos I (2017) The distinct function of Tep2 and Tep6 in the immune defense of Drosophila melanogaster against the pathogen Photorhabdus. Virulence 8(8):1668–1682.
  113. Silveira H, Gabriel A, Ramos S, Palma J, Felix R, Custódio A, Collins LV (2012) CpG-containing oligodeoxynucleotides increases resistance of Anopheles mosquitoes to plasmodium infection. Insect Biochem Mol Biol 42(10):758–765. Scholar
  114. Strand MR (2008) The insect cellular immune response. Insect Sci 15(1):1–14. Scholar
  115. Strand MR, Burke GR (2012) Polydnaviruses as symbionts and gene delivery systems. PLoS Pathog 8(7):e1002757. Scholar
  116. Su X-D, Gastinel LN, Vaughn DE, Faye I, Poon P, Bjorkman PJ (1998) Crystal structure of hemolin: a horseshoe shape with implications for homophilic Adhesion. Science 281(5379):991. Scholar
  117. Telfer WH, Kunkel JG (1991) The function and evolution of insect storage hexamers. Annu Rev Entomol 36(1):205–228. Scholar
  118. Theopold U, Schmidt O, Söderhäll K, Dushay MS (2004) Coagulation in arthropods: defence, wound closure and healing. Trends Immunol 25(6):289–294. Scholar
  119. Tokunaga F, Yamada M, Miyata T, Ding YL, Hiranaga-Kawabata M, Muta T, Iwanaga S, Ichinose A, Davie EW (1993) Limulus hemocyte transglutaminase. Its purification and characterization, and identification of the intracellular substrates. J Biol Chem 268(1):252–261Google Scholar
  120. Troha K, Buchon N (2019) Methods for the study of innate immunity in Drosophila melanogaster. Developmental Biology, Wiley Interdisciplinary Reviews e344.
  121. Tsuzuki S, Ochiai M, Matsumoto H, Kurata S, Ohnishi A, Hayakawa Y (2012) Drosophila growth-blocking peptide-like factor mediates acute immune reactions during infectious and non-infectious stress. Scient Rep 2:210–210.
  122. Vavricka C, Christensen B, Li J (2010) Melanization in living organisms: a perspective of species evolution. Protein Cell 1(9):830–841. Scholar
  123. Vavricka CJ, Han Q, Mehere P, Ding H, Christensen BM, Li J (2014) Tyrosine metabolic enzymes from insects and mammals: a comparative perspective. Insect Sci 21(1):13–19. Scholar
  124. Veillard F, Troxler L, Reichhart J-M (2016) Drosophila melanogaster clip-domain serine proteases: structure, function and regulation. Biochimie 122:255–269. Scholar
  125. Volz J, Müller H-M, Zdanowicz A, Kafatos FC, Osta MA (2006) A genetic module regulates the melanization response of Anopheles to plasmodium. Cell Microbiol 8(9):1392–1405. Scholar
  126. Wang Y, Jiang H, Cheng Y, An C, Chu Y, Raikhel AS, Zou Z (2017) Activation of Aedes aegypti prophenoloxidase-3 and its role in the immune response against entomopathogenic fungi. Insect Mol Biol 26(5):552–563. Scholar
  127. Wang Y, Jiang H, R. Kanost M (1999) Biological activity of Manduca sexta paralytic and plasmatocyte spreading peptide and primary structure of its hemolymph precursor. Insect Biochem Mol Biol 29 (12):1075–1086.
  128. Wang Y, Lu Z, Jiang H (2014) Manduca sexta proprophenoloxidase activating proteinase-3 (PAP3) stimulates melanization by activating proPAP3, proSPHs, and proPOs. Insect Biochem Mol Biol 50:82–91. Scholar
  129. Wang Z, Wilhelmsson C, Hyrsl P, Loof TG, Dobes P, Klupp M, Loseva O, Mörgelin M, Iklé J, Cripps RM, Herwald H, Theopold U (2010) Pathogen entrapment by Transglutaminase—a conserved early innate immune mechanism. PLoS Pathog 6(2):e1000763. Scholar
  130. Waterhouse R, Povelones M, Christophides G (2010) Sequence-structure-function relations of the mosquito leucine-rich repeat immune proteins. BMC Genom 11(1):531CrossRefGoogle Scholar
  131. Whitten MMA, Coates CJ (2017) Re-evaluation of insect melanogenesis research: views from the dark side. Pigment Cell Melanoma Res 30(4):386–401. Scholar
  132. Whitten MMA, Tew IF, Lee BL, Ratcliffe NA (2004) A novel role for an insect apolipoprotein (Apolipophorin III) in β-1,3-Glucan pattern recognition and cellular encapsulation reactions. J Immunol 172(4):2177–2185CrossRefGoogle Scholar
  133. Wigglesworth VB (1937) Wound healing in an insect (Rhodnius Prolixus Hemiptera). J Exp Biol 14(3):364–381Google Scholar
  134. Wynant N, Duressa TF, Santos D, Van Duppen J, Proost P, Huybrechts R, Vanden Broeck J (2014) Lipophorins can adhere to dsRNA, bacteria and fungi present in the hemolymph of the desert locust: A role as general scavenger for pathogens in the open body cavity. J Insect Physiol 64:7–13. Scholar
  135. Yang F, Wang Y, He Y, Jiang H (2016) In search of a function of Manduca sexta hemolymph protease-1 in the innate immune system. Insect Biochem Mol Biol 76:1–10. Scholar
  136. Yassine H, Kamareddine L, Chamat S, Christophides GK, Osta MA (2014) A Serine protease homolog negatively regulates TEP1 consumption in systemic infections of the malaria vector Anopheles gambiae. J Innate Immun 6(6):806–818. Scholar
  137. Yassine H, Kamareddine L, Osta MA (2012) The mosquito melanization response is implicated in defense against the entomopathogenic fungus beauveria bassiana. PLoS Path 8(11):e1003029. Scholar
  138. Yokoyama H, Yokoyama T, Yuasa M, Fujimoto H, Sakudoh T, Honda N, Fugo H, Tsuchida K (2013) Lipid transfer particle from the silkworm, Bombyx mori, is a novel member of the apoB/large lipid transfer protein family. J Lipid Res 54(9):2379–2390. Scholar
  139. Yu X-Q, Kanost MR (2000) Immulectin-2, a Lipopolysaccharide-specific Lectin from an Insect, Manduca sexta, Is induced in response to gram-negative bacteria. J Biol Chem 275(48):37373–37381. Scholar
  140. Zhang G, Lu Z-Q, Jiang H, Asgari S (2004) Negative regulation of prophenoloxidase (proPO) activation by a clip-domain serine proteinase homolog (SPH) from endoparasitoid venom. Insect Biochem Mol Biol 34(5):477–483CrossRefGoogle Scholar
  141. Zhang X, An C, Sprigg K, Michel K (2016) CLIPB8 is part of the prophenoloxidase activation system in Anopheles gambiae mosquitoes. Insect Biochem Mol Biol 71:106–115. Scholar
  142. Zhang X, He Y, Cao X, Gunaratna RT, Chen Y-r, Blissard G, Kanost MR, Jiang H (2015) Phylogenetic analysis and expression profiling of the pattern recognition receptors: insights into molecular recognition of invading pathogens in Manduca sexta. Insect Biochem Mol Biol 62:38–50. Scholar
  143. Zhang Y, Dong Z, Liu S, Yang Q, Zhao P, Xia Q (2012) Identification of novel members reveals the structural and functional divergence of lepidopteran-specific Lipoprotein_11 family. Funct Integr Genomics 12(4):705–715. Scholar
  144. Zhao P, Wang GH, Dong ZM, Duan J, Xu PZ, Cheng TC, Xiang ZH, Xia QY (2010) Genome-wide identification and expression analysis of serine proteases and homologs in the silkworm Bombyx mori. BMC Genom 11:405. Scholar
  145. Zou Z, Shin SW, Alvarez KS, Kokoza V, Raikhel AS (2010) Distinct melanization pathways in the mosquito aedes aegypti. Immunity 32(1):41–53. Scholar

Copyright information

© Springer Nature Switzerland AG 2020

Authors and Affiliations

  1. 1.Department of ChemistryUniversity of GeorgiaAthensUSA

Personalised recommendations