Fish Physiology and Biochemistry

, Volume 39, Issue 2, pp 131–142 | Cite as

Molecular cloning and expression analysis of major histocompatibility complex class IIB gene of the Whitespotted bambooshark (Chiloscyllium plagiosum)

  • Qian Ma
  • Yong-Quan Su
  • Jun WangEmail author
  • Zhi-Meng Zhuang
  • Qi-Sheng Tang


Major histocompatibility complex (MHC) plays an important role in the immune response to antigenic peptides in vertebrates. In this study, the full length of MHC IIB cDNA was isolated from the Whitespotted bambooshark (Chiloscyllium plagiosum) by homology cloning, and the rapid amplification of cDNA ends polymerase chain reaction. As a result, the MHC IIB cDNA is 1,407 bp, which contains an open reading frame (ORF) of 831 bp encoding a protein of 276 amino acids. Furthermore, seven alleles of the complete MHC IIB ORF were detected and the variable sites were mainly located in the immunoglobulin-like (β2) region. Tissue distribution analysis showed that MHC IIB can be detected in all the ten tissues examined, with the highest expression in the spleen and gill. Challenge of C. plagiosum with the pathogenic bacteria, Vibrio harveyi, resulted in significant changes in the expression of MHC IIB mRNA in the three immune-related tissues (gill, liver and spleen). These results show that the MHC IIB plays an important role in response to bacterial infection in elasmobranches.


Chiloscyllium plagiosum MHC IIB Sequence analysis mRNA expression Vibrio harveyi 



The research was financially supported by the National Natural Science Foundation of China (Grant No.: 40876080).


  1. Alsina M, Blanch AR (1994) A set of keys for biochemical identification of environmental Vibrio species. J Appl Microbiol 76(1):79–85CrossRefGoogle Scholar
  2. Anonymous (1985) Validation of the publication of new names and new combinations previously effectively published outside the IJSB. Int J Syst Bacteriol (List no 17) 35:223–224CrossRefGoogle Scholar
  3. Bartl S (2001) New major histocompatibility complex class IIB genes from nurse shark. Adv Exp Med Biol 484:1–11PubMedCrossRefGoogle Scholar
  4. Bartl S, Weissman IL (1994) Isolation and characterization of major histocompatibility complex class IIB genes from the nurse shark. Proc Nat Acad Sci 91(1):262–266PubMedCrossRefGoogle Scholar
  5. Bertone S, Gili C, Moizo A, Calegari L (1996) Vibrio carchariae associated with a chronic skin ulcer on a shark, Carcharhinus plumbeus (Nardo). J Fish Dis 19(6):429–434CrossRefGoogle Scholar
  6. Buonocore F, Randelli E, Casani D, Costantini S, Facchiano A, Scapigliati G, Stet RJM (2007) Molecular cloning, differential expression and 3D structural analysis of the MHC class-II [beta] chain from sea bass (Dicentrarchus labrax L.). Fish Shellfish Immunol 23(4):853–866PubMedCrossRefGoogle Scholar
  7. Cano-Gomez A, Bourne DG, Hall MR, Owens L, Høj L (2009) Molecular identification, typing and tracking of Vibrio harveyi in aquaculture systems: current methods and future prospects. Aquaculture 287(1–2):1–10CrossRefGoogle Scholar
  8. Cho J, Kim Y (2002) Sharks: a potential source of antiangiogenic factors and tumor treatments. Mar Biotechnol 4(6):521–525PubMedCrossRefGoogle Scholar
  9. Compagno LJV (2001) Sharks of the world: an annotated and illustrated catalogue of shark species known to date, volume 2. Bullhead, mackerel and carpet sharks (Heterodontiformes, Lamniformes and Orectolobiformes), FAO, RomeGoogle Scholar
  10. Cong M, Song LS, Wang LL, Zhao JM, Qiu LM, Li L, Zhang H (2008) The enhanced immune protection of Zhikong scallop Chlamys farreri on the secondary encounter with Listonella anguillarum. Comp Biochem Physiol B Biochem Mol Biol 151(2):191–196PubMedCrossRefGoogle Scholar
  11. Cosson P, Bonifacino JS (1992) Role of transmembrane domain interactions in the assembly of class II MHC molecules. Science 258(5082):659–662PubMedCrossRefGoogle Scholar
  12. Dooley H, Flajnik MF (2005) Shark immunity bites back: affinity maturation and memory response in the nurse shark, Ginglymostoma cirratum. Eur J Immunol 35(3):936–945PubMedCrossRefGoogle Scholar
  13. Espenes A, Press CM, Reitan LJ, Landsverk T (1996) The trapping of intravenously injected extracellular products from Aeromonas salmonicidain head kidney and spleen of vaccinated and nonvaccinated Atlantic salmon, Salmo salar L. Fish Shellfish Immunol 6(6):413–426CrossRefGoogle Scholar
  14. Fange R, Mattisson A (1981) The lymphomyeloid (hemopoietic) system of the Atlantic nurse shark, Ginglymostoma cirratum. Biol Bull 160(2):240–249CrossRefGoogle Scholar
  15. Figueroa F, Mayer WE, Sültmann H, O’hUigin C, Tichy H, Satta Y, Takezaki N, Takahata N, Klein J (2000) Mhc class II B gene evolution in East African cichlid fishes. Immunogenetics 51(7):556–575PubMedCrossRefGoogle Scholar
  16. Graser R, O’hUigin C, Vincek V, Meyer A, Klein J (1996) Trans-species polymorphism of class II Mhc loci in danio fishes. Immunogenetics 44(1):36–48PubMedCrossRefGoogle Scholar
  17. Grimes D, Colwell R, Stemmler J, Hada H, Maneval D, Hetrick F, May E, Jones R, Stoskopf M (1984) Vibrio species as agents of elasmobranch disease. Helgol Mar Res 37(1):309–315Google Scholar
  18. Grimes DJ, Jacobs D, Swartz D, Brayton P, Colwell RR (1993) Numerical taxonomy of gram-negative, oxidase-positive rods from carcharhinid sharks. Int J Syst Evol Microbiol 43(1):88–98Google Scholar
  19. Grimholt U, Larsen S, Nordmo R, Midtlyng P, Kjoeglum S, Storset A, Saebø S, Stet RJM (2003) MHC polymorphism and disease resistance in Atlantic salmon (Salmo salar); facing pathogens with single expressed major histocompatibility class I and class II loci. Immunogenetics 55(4):210–219PubMedCrossRefGoogle Scholar
  20. Hansen JD, La Patra S (2002) Induction of the rainbow trout MHC class I pathway during acute IHNV infection. Immunogenetics 54(9):654–661PubMedCrossRefGoogle Scholar
  21. Hashimoto K, Nakanishi T, Kurosawa Y (1992) Identification of a shark sequence resembling the major histocompatibility complex class I alpha 3 domain. Proc Nat Acad Sci 89(6):2209–2212PubMedCrossRefGoogle Scholar
  22. Hordvik I, Grimholt U, Fosse VM, Lie, Endresen C (1993) Cloning and sequence analysis of cDNAs encoding the MHC class II β chain in Atlantic salmon (Salmo salar). Immunogenetics 37(6):437–441PubMedCrossRefGoogle Scholar
  23. Kita-Tsukamoto K, Oyaizu H, Nanba K, Simidu U (1993) Phylogenetic relationships of marine bacteria, mainly members of the family Vibrionaceae, determined on the basis of 16S rRNA sequences. Int J Syst Evol Microbiol 43(1):8–19Google Scholar
  24. Könighuang RL, Germain RN (1992) MHC class II interaction with CD4 mediated by a region analogous to the MHC class I binding site for CD8. Nature 356:796–798CrossRefGoogle Scholar
  25. Koppang EO, Lundin M, Press CM, Rønningen K, Lie Ø (1998) Differing levels of Mhc class II β chain expression in a range of tissues from vaccinated and non-vaccinated Atlantic salmon (Salmo salarL.). Fish Shellfish Immunol 8(3):183–196CrossRefGoogle Scholar
  26. Koppang EO, Dannevig BH, Lie O, Ronningen K, Press CML (1999) Expression of Mhc class I and II mRNA in a macrophage-like cell line (SHK-1) derived from Atlantic salmon, Salmo salar L., head kidney. Fish Shellfish Immunol 9(6):473–489CrossRefGoogle Scholar
  27. Librado P, Rozas J (2009) DnaSP v5: a software for comprehensive analysis of DNA polymorphism data. Bioinformatics 25:1451–1452PubMedCrossRefGoogle Scholar
  28. Livak K, Schmittgen T (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2T−ΔΔC method. Methods 25(4):402–408PubMedCrossRefGoogle Scholar
  29. Ma Q, Hong J, Jiang Q, Su Y, Wang J (2010) Effects of Vibrio harveyi on the activities of four enzymes in Chiloscyllium plagiosum. J Oceanog (in Taiwan Strait) 29(2):196–204Google Scholar
  30. Miller KM, Withler RE (1996) Sequence analysis of a polymorphic Mhc class II gene in Pacific salmon. Immunogenetics 43(6):337–351PubMedCrossRefGoogle Scholar
  31. Morrison RN, Koppang EO, Hordvik I, Nowak BF (2006) MHC class II+ cells in the gills of Atlantic salmon (Salmo salar L.) affected by amoebic gill disease. Vet Immunol Immunopathol 109(3–4):297–303PubMedCrossRefGoogle Scholar
  32. Ottová E, Šimková A, Martin JF, Goüy de Bellocq J, Gelnar M, Allienne JF, Morand S (2005) Evolution and trans-species polymorphism of MHC class II [beta] genes in cyprinid fish. Fish Shellfish Immunol 18(3):199–222PubMedCrossRefGoogle Scholar
  33. Pedersen K, Verdonck L, Austin B, Austin DA, Blanch AR, Grimont PAD, Jofre J, Koblavi S, Larsen JL, Tiainen T (1998) Taxonomic evidence that Vibrio carchariae Grimes et al. 1985 is a junior synonym of Vibrio harveyi (Johnson and Shunk 1936) Baumann et al. 1981. Int J Syst Evol Microbiol 48(3):749–758Google Scholar
  34. Pettey CL, McKinney EC (1983) Temperature and cellular regulation of spontaneous cytotoxicity in the shark. Eur J Immunol 13(2):133–138PubMedCrossRefGoogle Scholar
  35. Press CML, Dannevig BH, Landsverk T (1994) Immune and enzyme histochemical phenotypes of lymphoid and nonlymphoid cells within the spleen and head kidney of Atlantic salmon (Salmo salar L.). Fish Shellfish Immunol 4(2):79–93CrossRefGoogle Scholar
  36. Rumfelt L, McKinney E, Taylor E, Flajnik M (2002) The development of primary and secondary lymphoid tissues in the nurse shark Ginglymostoma cirratum: B-cell zones precede dendritic cell immigration and T-cell zone formation during ontogeny of the spleen. Scand J Immunol 56(2):130–148PubMedCrossRefGoogle Scholar
  37. Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4(4):406–425PubMedGoogle Scholar
  38. Stet RJM, Dixon B, Van Erp SHM, Van Lierop MJC, Rodrigues PNS, Egberts E (1996) Inference of structure and function of fish major histocompatibility complex (MHC) molecules from expressed genes. Fish Shellfish Immunol 6(4):305–318CrossRefGoogle Scholar
  39. Xu TJ, Chen SL, Ji XS, Sha ZX (2009a) Molecular cloning, genomic structure, polymorphism and expression analysis of major histocompatibility complex class IIA and IIB genes of half-smooth tongue sole (Cynoglossus semilaevis). Fish Shellfish Immunol 27(2):192–201PubMedCrossRefGoogle Scholar
  40. Xu XJ, Xu B, Wang J, Su YQ, Zhang ZW, Chen X (2009b) Histopathological Studies on Pseudosciaena crocea Artificially Challenged with Vibrio harveyi. J Xiamen Univ (Nat Sci) 48(2):281–286Google Scholar

Copyright information

© Springer Science+Business Media B.V. 2012

Authors and Affiliations

  • Qian Ma
    • 1
    • 2
  • Yong-Quan Su
    • 2
  • Jun Wang
    • 2
    Email author
  • Zhi-Meng Zhuang
    • 1
  • Qi-Sheng Tang
    • 1
  1. 1.Key Laboratory for Fishery Resources and Eco-environment, Yellow Sea Fisheries Research InstituteChinese Academy of Fishery SciencesQingdaoChina
  2. 2.College of Oceanography and Environmental ScienceXiamen UniversityXiamenChina

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