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Induction of phenoloxidase and other immunological activities in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Lipopolysaccharide (LPS)

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Abstract

The knowledge concerning the humoral immunity is scarce in amphioxus Branchiostoma belcheri. This study measured the humoral parameters including phenoloxidase (PO) activity, lysozyme activity, antimicrobial activity, microbial agglutinin, and hemagglutinin in amphioxus humoral fluids before and after lipopolysaccharide (LPS) challenge. Humoral fluids from unchallenged Branchiostoma belcheri (B. belcheri) had PO activity, lysozyme, antimicrobial, microbial agglutinating, and hemagglutinating activities, which may represent part of the baseline level of innate immunity in this organism. After challenge with LPS, many humoral parameters were all increased significantly including the PO activity, lysozyme activity, growth-inhibiting activities against Gram-negative bacteria Escherichia coli (E. coli) and Vibrio alginolyticus (V. alginolyticus), growth-inhibiting activities against Gram-positive bacteria Staphylococcus aureus (S. aureus) and Bacillus subtilis (B. subtilis), microbial agglutinating activities against Micrococcus lysodeikticus (M. lysodeikticus), B. subtilis, and S. aureus, and hemagglutinating activities against rabbit and human A and O erythrocytes. In contrast, the agglutinating activities against V. harveyi and E. coli and the hemagglutinating activity against human B erythrocytes in the humoral fluids were reduced in response to LPS challenge. It appears that the humoral fluids of B. belcheri contain components that are able to differentiate different microbes and different human blood cell types.

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References

  • Aladaileh S, Nair SV, Raftos DA (2007) Induction of phenoloxidase and other immunological activities in Sydney rock oysters challenged with microbial pathogen-associate molecular patterns. Fish Shellfish Immunol 23(6):1196–1208

    Article  CAS  Google Scholar 

  • Alam M, Miyoshi S, Tomochika K, Shinodo S (1997) Haemagglutination is a novel biological function of lipopolysaccharide (LPS) as seen with the Vibrio cholerae 0139 LPS. Clin Diag Lab Immunol 4(5):604–606

    CAS  Google Scholar 

  • Asokan R, Arumugam M, Mullainadhan P (1997) Activation of prophenoloxidase in the plasma and haemocytes of the marine mussel, Perna viridis Linnaeus. Dev Comp Immunol 21:1–12

    Article  CAS  Google Scholar 

  • Berczi I (1998) Neurohormonal host defense in endotoxin shock. Ann NY Acad Sci 84:787–802

    Article  Google Scholar 

  • Cannon JP, Haire RN, Litman GW (2002) Identification of diversified genes that contain immunoglobulin-like variable regions in a protochordate. Nat Immunol 3:1200–1207

    Article  CAS  Google Scholar 

  • Cardenas W, Dankert JR (1997) Phenoloxidase specific activity in the red swamp crayfish Procambarus clarkii. Fish Shellfish Immunol 7:283–295

    Article  Google Scholar 

  • Cardenas W, Dankert JR, Jenkins JA (2004) Flow cytometric analysis of crayfish haemocytes activated by lipopolysaccharides. Fish Shellfish Immunol 17:223–233

    Article  CAS  Google Scholar 

  • Charalambidis ND, Foukas LC, Marmaras VJ (1996) Covalent association of lipopolysaccharide at the hemocyte surface of insects is an initial step for its internalization–Protein-tyrosine phosphorylation requirement. Eur J Biochem 236(1):200–206

    Article  CAS  Google Scholar 

  • Coles JA, Pipe RK (1994) Phenoloxidase activity in the haemolymph and haemocytes of the marine mussel Mytilus edulis. Fish Shellfish Immunol 4:337–352

    Article  Google Scholar 

  • Erridge C, Bennett-Guerrero E, Poxton IR (2002) Structure and function of lipopolysaccharide. Microbiol Infect 4(8):837–841

    Article  CAS  Google Scholar 

  • Horowitz NH, Shen SC (1952) Neurospora tyrosinase. J Biol Chem 197:513–520

    CAS  PubMed  Google Scholar 

  • Ingram GA, Alexander JB (1980) The immune response of the brown trout Salmo trutta to lipopolysaccharide. J Fish Biol 16:181–197

    Article  CAS  Google Scholar 

  • Jakobsen RA, Gutierez MA, Wergeland HI (1999) Antibodies to Aeromonas salmonicida lipopolysaccharide and cell wall antigens in rabbit and salmon immune sera. Fish Shellfish Immunol 9(8):609–620

    Article  Google Scholar 

  • Jin P, Hu J, Qian J, Chen L, Xu X, Ma F (2012) Identification and characterization of a putative lipopolysaccharide-induced TNF-α factor (LITAF) gene from Amphioxus (Branchiostoma belcheri): an insight into the innate immunity of Amphioxus and the evolution of LITAF. Fish Shellfish Immunol (in press). doi:10.1016/j.fsi.2012.03.030

    Article  CAS  Google Scholar 

  • Kato Y, Motoi Y, Taniai K, Kadono-Okuda K, Yamamoto M, Higashino Y, Shimabukuro M, Chowdhury S, Xu J, Sugiyama M et al (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

    Article  CAS  Google Scholar 

  • Korner P, Schmid-Hempel P (2004) In vivo dynamics of an immune response in the bumble bee Bombus terrestris. J Invertebr Pathol 87(1):59–66

    Article  CAS  Google Scholar 

  • Laemmli UK (1970) Cleavage of structural proteins during assembly of the head of bacteriophage T4. Nature 227:680–685

    Article  CAS  Google Scholar 

  • Li ZM, Zhang SC, Wang CF, Pang QX (2008) Complement-mediated killing of Vibrio species by the humoral fluids of amphioxus Branchiostoma belcheri: implications for a dual role of O-antigens in the resistance to bactericidal activity. Fish Shellfish Immunol 24:215–222

    Article  Google Scholar 

  • Liang YJ, Zhang SC, Lun LM, Han L (2006) Presence and localization of antithrombin and its regulation after acute lipopolysaccharide exposure in amphioxus, with implications for the origin of vertebrate liver. Cell Tissue Res 323:537–541

    Article  CAS  Google Scholar 

  • Liang YJ, Zhang SC, Wang ZP (2009) Alternative complement activity in the egg cytosol of amphioxus Branchiostoma Belcheri: evidence for the defense role of maternal complement components. PLoS One 4(1):e4234

    Article  Google Scholar 

  • Liang Y, Pan A, Zhang S, Zhang Y, Liu M (2011) Cloning, distribution and primary immune characteristics of amphioxus alpha-2 macroglobulin. Fish Shellfish Immunol 31(6):963–969

    Article  CAS  Google Scholar 

  • Ling E, Yu XQ (2006) Cellular encapsulation and melanization are enhanced by immulectins, pattern recognition receptors from the tobacco hornworm Manduca sexta. Dev Comp Immunol 30:289–299

    Article  CAS  Google Scholar 

  • Liu N, Zhang S, Liu Z (2007) Characterization and expression of gamma-interferon-inducible lysosomal thiol reductase (GILT) gene in amphioxus Branchiostoma belcheri with implications for GILT in innate immune response. Mol Immunol 44(10):2631–2637

    Article  CAS  Google Scholar 

  • McCabe WR (1980) Endotoxin: microbiological, chemical, pathophysiological and clinical correlations. Sem Infect Dis 3:38–88

    CAS  Google Scholar 

  • Minagawa S, Hikima J, Hirono I, Aoki T, Mori H (2001) Expression of Japanese flounder c-type lysozyme cDNA in insect cells. Dev Comp Immunol 25:439–445

    Article  CAS  Google Scholar 

  • Nayak SK, Swain P, Nanda PK, Dash S, Shukla S, Meher PK, Maiti NK (2007) Effect of endotoxin on the immunity of Indian major carp Labeo rohita. Fish Shellfish Immunol 24(3):394–399

    PubMed  Google Scholar 

  • Nayak SK, Swain P, Nanda PK, Dash S, Shukla S, Meher PK et al (2008) Effect of endotoxin on the immunity of Indian major carp, Labeo rohita. Fish Shellfish Immunol 24:394–399

    Article  CAS  Google Scholar 

  • Nayak SK, Swain P, Nanda PK, Mohapatra D, Behera T (2011) Immunomodulating potency of lipopolysaccharides (LPS) derived from smooth type of bacterial pathogens in Indian major carp. Vet Microbiol 151:413–417

    Article  CAS  Google Scholar 

  • Nellaiappan K, Vinayagam A (1986) A rapid method for detection of tyrosinase activity in electrophoresis. Stain Technol 61:269–272

    Article  CAS  Google Scholar 

  • Novoa B, Bowman TV, Zon L, Figueras A (2009) LPS response and tolerance in the zebrafish (Danio rerio). Fish Shellfish Immunol 26:326–331

    Article  CAS  Google Scholar 

  • Okumura T (2007) Effects of lipopolysaccharide on gene expression of antimicrobial peptides (penaeidins and crustin), serine proteinase and prophenoloxidase in haemocytes of the Pacific white shrimp, Litopenaeus vannamei. Fish Shellfish Immunol 22:68–76

    Article  CAS  Google Scholar 

  • Pang QX, Zhang SC, Wang CF, Shi XD, Sun YN (2004) Presence of prophenoloxidase in the humoral fluid of amphioxus Branchiostoma belcheri tsingtauense. Fish Shellfish Immunol 17:477–487

    Article  CAS  Google Scholar 

  • Pang QX, Zhang SC, Shi XD, Su F, Wu D (2005) Purification and characterisation of phenoloxidase from amphioxus Branchiostoma belcheri tsingtauense. Fish Shellfish Immunol 19:139–148

    Article  CAS  Google Scholar 

  • Pang QX, Zhang SC, Liu XM, Wu D (2006) Humoral immune responses of amphioxus Branchiostoma belcheri to challenge with Escherichia coli. Fish Shellfish Immunol 21:139–145

    Article  CAS  Google Scholar 

  • Pang QX, Zhang SC, Zhao BS (2009) Induction of phenoloxidases in the humoral fluids of amphioxus Branchiostoma belcheri by Vibrio alginolyticus and Escherichia coli. Fish Shellfish Immunol 26:669–671

    Article  CAS  Google Scholar 

  • Pang QX, Zhang SC, Zhao BS (2010) Immune parameters in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Vibrio alginolyticus. Fish Shellfish Immunol 28:232–234

    Article  CAS  Google Scholar 

  • Paulsen SM, Lunde H, Engstad RE, Robertsen B (2003) In vivo effects of beta-glucan and LPS on regulation of lysozyme activity and mRNA expression in Atlantic salmon (Salmo salar L.). Fish Shellfish Immunol 14:39–54

    Article  CAS  Google Scholar 

  • Ratcliffe NA, Brookman JL, Rowley AF (1991) Activation of the prophenoloxidase cascade and initiation of nodule formation in locusts by bacterial lipopolysaccharides. Dev Comp Immunol 15:33–39

    Article  CAS  Google Scholar 

  • Rees JA, Moniatte M, Bulet P (1997) Novel antibacterial peptides isolated from a European bumblebee, Bombus pascuorum (Hymenoptera, Apoidea). Insect Biochem Mol Biol 27:413–422

    Article  CAS  Google Scholar 

  • Rietschel ET, Kirikae T, Schade FU, Mamat U, Schmidt G, Loppnow H, Ulmer AJ, Zahringer U, Seydel U, Di Padova F (1994) Bacterial endotoxin: molecular relationships of structure to activity and function. FASEB J 8:217–225

    Article  CAS  Google Scholar 

  • Sato A, Mayer WE, Klein J (2003) A molecule bearing an immunoglobulin-like V region of the CTX subfamily in amphioxus. Immunogenetics 55:423–427

    Article  CAS  Google Scholar 

  • Selvaraj V, Sampath K, Sekar V (2004) Extraction and characterization of lipopolysaccharide from Aeromonas hydrophila and its effects on survival and hematology of the carp Cyprinus carpio. Asian Fish Sci 17:163–173

    Google Scholar 

  • Selvaraj V, Sampath K, Sekar V (2009) Administration of lipopolysaccharide increases specific and non-specific immune parameters and survival in carp (Cyprinus carpio) infected with Aeromonas hydrophila. Aquaculture 286:176–183

    Article  CAS  Google Scholar 

  • Skugor S, Skugor A, Todorcevic M, Torgersen J, Ruyter B, Krasnov A (2010) Exposure to lipopolysaccharide induces immune genes in cultured preadipocytes of Atlantic salmon. Fish Shellfish Immunol 29:817–824

    Article  CAS  Google Scholar 

  • Soderhall K, Hall L (1984) Lipopolysaccharide-induced activation of prophenoloxidase activating system in crayfish haemocyte lysate. Biochim Biophys Acta 797:99–104

    Article  Google Scholar 

  • Solem ST, Jorgensen JB, Robertsen B (1995) Stimulation of respiratory burst and phagocytic activity in Atlantic salmon (Salmo salar L.) macrophages by lipopolysaccharide. Fish Shellfish Immunol 5:475–491

    Article  Google Scholar 

  • Sritunyalucksana K, Sithisarn P, Withayachumnarnkul B, Flegel TW (1999) Activation of prophenoloxidase, agglutinin and antibacterial activity in haemolymph of the black tiger prawn, Penaeus monodon, by immunostimulants. Fish Shellfish Immunol 9:21–30

    Article  Google Scholar 

  • Swain P, Nayak SK, Nanda PK, Dash S (2008) Biological effects of bacterial lipopolysaccharide (endotoxin) in fish: a review. Fish Shellfish Immunol 25(3):191–201

    Article  CAS  Google Scholar 

  • Wang CF, Zhang SC, Luo Y, Xu TT (2002) Presence and induction by bacteria, of D-galactoside-specific lectins in the humoral fluids of amphioxus Branchiostoma belcheri tsingtauense. Inflammopharmacology 9:241–248

    Article  CAS  Google Scholar 

  • Yan J, Wang J, Zhao Y, Zhang J, Bai C, Zhang C, Zhang C, Li K, Zhang H, Du X, Feng L (2012) Identification of an amphioxus intelectin homolog that preferably agglutinates gram-positive over gram-negative bacteria likely due to different binding capacity to LPS and PGN. Fish Shellfish Immunol (in press). doi:10.1016/j.fsi.2012.03.023

    Article  CAS  Google Scholar 

  • Yu C, Dong M, Wu X, Li S, Huang S, Su J et al (2005) Genes “waiting” for recruitment by the adaptive immune system: the insights from amphioxus. J Immunol 174:3493–3500

    Article  CAS  Google Scholar 

  • Zhang SC, Sun YN, Pang QX, Shi XD (2005) Hemagglutinating and antibacterial activities of vitellogenin. Fish Shellfish Immunol 19:93–95

    Article  CAS  Google Scholar 

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Acknowledgments

We thank Prof. Shicui Zhang for his help and advice in this study. This work was supported by the National Natural Science Foundation of China (Grant No. 31172074; Grant No. 30700071), the Shandong Provincial Natural Science Foundation (Grant No. ZR2010CL002), Young Teacher Support Program in Shandong University of Technology, and the National High Technology and Development Program (Grant No. 2008AA09Z411).

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Authors and Affiliations

  1. School of Life Sciences, Shandong University of Technology, Zibo, 255049, People’s Republic of China

    Qiuxiang Pang, Bosheng Zhao & Huanhuan Sun

  2. Department of Nephrology, Affiliated Hospital of Medical College, Qingdao University, Qingdao, 266003, People’s Republic of China

    Xuemei Liu

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  1. Qiuxiang Pang
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Correspondence to Bosheng Zhao.

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Pang, Q., Liu, X., Zhao, B. et al. Induction of phenoloxidase and other immunological activities in the humoral fluids of amphioxus Branchiostoma belcheri challenged with Lipopolysaccharide (LPS). Fish Physiol Biochem 38, 1835–1842 (2012). https://doi.org/10.1007/s10695-012-9680-7

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  • DOI: https://doi.org/10.1007/s10695-012-9680-7

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