Advertisement

Molecular Biology Reports

, Volume 42, Issue 2, pp 431–439 | Cite as

The CD8α gene in duck (Anatidae): cloning, characterization, and expression during viral infection

  • Qi Xu
  • Yang Chen
  • Wen Ming Zhao
  • Zheng Yang Huang
  • Xiu Jun Duan
  • Yi Yu Tong
  • Yang Zhang
  • Xiu Li
  • Guo Bin Chang
  • Guo Hong ChenEmail author
Article

Abstract

Cluster of differentiation 8 alpha (CD8α) is critical for cell-mediated immune defense and T-cell development. Although CD8α sequences have been reported for several species, very little is known about CD8α in ducks. To elucidate the mechanisms involved in the innate and adaptive immune responses of ducks, we cloned CD8α coding sequences from domestic, Muscovy, Mallard, and Spotbill ducks using reverse transcription polymerase chain reaction (RT-PCR). Each sequence consisted of 714 nucleotides and encoded a signal peptide, an IgV-like domain, a stalk region, a transmembrane region, and a cytoplasmic tail. We identified 58 nucleotide differences and 37 amino acid differences among the four types of duck; of these, 53 nucleotide and 33 amino acid differences were between Muscovy ducks and the other duck species. The CD8α cDNA sequence from domestic duck consisted of a 61-nucleotide 5′ untranslated region (UTR), a 714-nucleotide open reading frame, and an 849-nucleotide 3′ UTR. Multiple sequence alignments showed that the amino acid sequence of CD8α is conserved in vertebrates. RT-PCR revealed that expression of CD8α mRNA of domestic ducks was highest in the thymus and very low in the kidney, cerebrum, cerebellum, and muscle. Immunohistochemical analyses detected CD8α on the splenic corpuscle and periarterial lymphatic sheath of the spleen. CD8α mRNA in domestic ducklings was initially up-regulated, and then down-regulated, in the thymus, spleen, and liver after treatment with duck hepatitis virus type I (DHV-1) or the immunostimulant polyriboinosinic polyribocytidylic acid (poly I:C).

Keywords

CD8α Duck Gene cloning Gene expression Viral infection 

Notes

Acknowledgments

We are grateful to Prof. Peng DX and Dr. Chen SJ (Yangzhou University) for their suggestions and technical assistance. This work was supported financially by the National Natural Science Foundation of China (31101704), Natural Science Foundation of Jiangsu Province (BK20141275) and the Priority Academic Program Development of Jiangsu Higher Education Institutions (2011-137).

Conflict of interest

The authors have declared that no conflict of interest exists.

References

  1. 1.
    Huang YH, Li N, Burt DW, Wu F (2008) Genomic research and applications in the duck (Anas platyrhynchos). World Poult Sci J 64:329–341CrossRefGoogle Scholar
  2. 2.
    van Loenen MM, Hagedoorn RS, de Boer R, Falkenburg JH, Heemskerk MH (2013) Extracellular domains of CD8α and CD8ß subunits are sufficient for HLA class I restricted helper functions of TCR-engineered CD4(+) T cells. PLoS ONE 30:e65212CrossRefGoogle Scholar
  3. 3.
    Abbas AK, Lichtman AH (2003) Cellular and molecular immunology, 5th edn. Saunders, PhiladelphiaGoogle Scholar
  4. 4.
    Hu Q, Pan Z, Deen S, Meng S, Zhang X, Zhang X, Jiao XA (2007) New alleles of chicken CD8α and CD3ε found in Chinese native and western breeds. Vet Immunol Immunopathol 120:223–233CrossRefPubMedGoogle Scholar
  5. 5.
    Tang XS, Wang L, Liao DW, Jiang YN, Xia C (2007) Cloning, expression and production of polyclonal antibodies in extracellular regions of chicken CD8α/β. J China Agr Univ 12:5–9Google Scholar
  6. 6.
    Liaw HJ, Chen WR, Huang YC, Tsai CW, Chang KC, Kuo CL (2007) Genomic organization of the chicken CD8 locus reveals a novel family of immunoreceptor genes. J Immuno 178:3023–3030CrossRefGoogle Scholar
  7. 7.
    Tregaskes CA, Kong FK, Paramithiotis E, Chen CL, Ratcliffe MJ, Davison TF, Young JR (1995) Identification and analysis of the expression of CD8αβ and CD8αα isoforms in chickens reveals a major TCR-γδ CD8αβ subset of intestinal intraepithelial lymphocytes. J Immunol 154:4485–4494PubMedGoogle Scholar
  8. 8.
    Kothlow S, Mannes NK, Schaerer B, Rebeski DE, Kaspers B, Schultz U (2005) Characterization of duck leucocytes by monoclonal antibodies. Dev Comp Immunol 29:733–748CrossRefPubMedGoogle Scholar
  9. 9.
    Fletcher OJ, Tan X, Cortes L (2012) Cost effective and time efficient measurement of CD4, CD8, major histocompatibility complex class II, and macrophage antigen expression in the lungs of chickens. Vet Immunol Immunopathol 146:225–236CrossRefPubMedGoogle Scholar
  10. 10.
    Reaiche GY (2008) Characterization of the events involved in the resolution of acute duck hepatitis B virus infection. Thesis of the University of Adelaide, AdelaideGoogle Scholar
  11. 11.
    Mancebo E, Moreno-Pelayo MA, Mencía A, de la Calle-Martín O, Allende LM, Sivadorai P, Kalaydjieva L, Bertranpetit J, Coto E, Calleja-Antolín S, Ruiz-Contreras J, Paz-Artal E (2008) Gly111Ser mutation in CD8A gene causing CD8 immunodeficiency is found in Spanish gypsies. Mol Immunol 45:479–484CrossRefPubMedGoogle Scholar
  12. 12.
    de la Calle-Martin O, Hernandez M, Ordi J, Casamitjana N, Arostegui JI, Caragol I, Ferrando M, Labrador M, Rodriguez-Sanchez JL, Espanol T (2001) Familial CD8 deficiency due to a mutation in the CD8 alpha gene. J Clin Invest 108:117–123PubMedCentralCrossRefPubMedGoogle Scholar
  13. 13.
    Jin X, Zhang W, Zhang W, Gu C, Cheng G, Hu X (2008) Identification and molecular analysis of the highly pathogenic duck hepatitis virus type I in Hubei province of China. Res Vet Sci 85:595–598CrossRefPubMedGoogle Scholar
  14. 14.
    Cheng HJ, Cheng AC, Wang MS, Chen XY (2007) Development of an indirect immunoperoxidase staining technique for the detection of duck hepatitis virus type 1. Vet Sci China 37:369–373Google Scholar
  15. 15.
    Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2−ΔΔCT method. Methods 25:402–408CrossRefPubMedGoogle Scholar
  16. 16.
    Hu XY, Cheng GF, Zhou SQ, Xiong DH (2000) Studies on histopathological changes of inoculated ducklings with DHV-1. J Huazhong Agric Univ 19:48–50Google Scholar
  17. 17.
    Gu CQ, Xie CQ, Hu XY, Zhang WP, Bi DR, Cheng GF (2012) Cytokine gene expression in the livers of ducklings infected with duck hepatitis virus-1 JX strain. Poult Sci 91:583–591CrossRefPubMedGoogle Scholar
  18. 18.
    Moebius U, Kober G, Griscelli AL, Hercend T, Meuer SC (1991) Expression of different CD8 isoforms on distinct human lymphocyte subpopulations. Eur J Immunol 21:1793–1800CrossRefPubMedGoogle Scholar
  19. 19.
    Mak TW, Rahemtulla A, Schilham M, Koh DR, Fung-Leung WP (1992) Generation of mutant mice lacking surface expression of CD4 or CD8 by gene targeting. J Autoimmun 5:55–59CrossRefPubMedGoogle Scholar
  20. 20.
    Nagarajan UM, O’Connell C, Rank RG (2004) Molecular characterization of guinea-pig (Cavia porcellus) CD8 alpha and CD8 beta cDNA. Tissue Antigens 63:184–189CrossRefPubMedGoogle Scholar
  21. 21.
    Leahy DJ, Axel R, Hendrickson WA (1992) Crystal structure of a soluble form of the human T-cell coreceptor CD8 at 2.6 Å resolution. Cell 68:1145–1162CrossRefPubMedGoogle Scholar
  22. 22.
    Gao GF, Tormo J, Gerth UC, Wyer JR, McMichael AJ, Stuart DI, Bell JI, Jones EY, Jakobsen BK (1997) Crystal structure of the complex between human CD8α and HLA-A2. Nature 387:630–634CrossRefPubMedGoogle Scholar
  23. 23.
    Tanabe M, Karaki S, Takiguchi M, Nakauchi H (1992) Antigen recognition by the T-cell receptor is enhanced by CD8 alpha-chain binding to the alpha 3 domain of MHC class I molecules, not by signaling via the cytoplasmic domain of CD8 alpha. Int Immunol 4:147–152CrossRefPubMedGoogle Scholar
  24. 24.
    Shaw AS, Chalupny J, Whitney JA, Hammond C, Amrein KE, Kavathas P, Sefton BM, Rose JK (1990) Short related sequences in the cytoplasmic domains of CD4 and CD8 mediate binding to the amino-terminal domain of the p56lck tyrosine protein kinase. Mol Cell Biol 10:1853–1862PubMedCentralPubMedGoogle Scholar
  25. 25.
    Barber EK, Dasgupta JD, Schlossman SF, Trevillyan JM, Rudd CE (1989) The CD4 and CD8 antigens are coupled to a protein-tyrosine kinase (p56lck) that phosphorylates the CD3 complex. Proc Natl Acad Sci 86:3277–3281PubMedCentralCrossRefPubMedGoogle Scholar
  26. 26.
    Louise GD, Sham VN, Elizabeth MD (2009) The marsupial CD8 gene locus: molecular cloning and expression analysis of the alpha and beta sequences in the gray short-tailed opossum (Monodelphis domestica) and the tammar wallaby (Macropus eugenii). Vet Immunol Immunopathol 129:14–27CrossRefGoogle Scholar
  27. 27.
    Simona P, Laura G, Francesco B (2009) Lymphocyte differentiation in sea bass thymus: CD4 and CD8-α gene expression studies. Fish Shellfish Immunol 27:50–56CrossRefGoogle Scholar
  28. 28.
    Yue H, Huang X, Yang FL, Li MY, Fan GC, Ma L, Tang C (2008) Development of real-time RT-PCR for detecting the expression level of CD4 and CD8 mRNA in chicken. Acta Vet Zootech Sin 39:784–790Google Scholar
  29. 29.
    Hansen JD, Zapata AG (1998) Lymphocyte development in fish and amphibians. Immunol Rev 166:199–220CrossRefPubMedGoogle Scholar
  30. 30.
    Zwollo P, Cole S, Bromage E, Kaattari S (2005) B-cell heterogeneity in the teleost kidney: evidence for a maturation gradient from anterior to posterior kidney. J Immunol 174:6608–6616CrossRefPubMedGoogle Scholar
  31. 31.
    Davidson GA, Lin SH, Secombes CJ, Ellis AE (1997) Detection of specific and ‘constitutive’ antibody secreting cells in the gills, head kidney and peripheral blood leucocytes of dab (Limanda limanda). Vet Immunol Immunopathol 58:363–374CrossRefPubMedGoogle Scholar
  32. 32.
    Anwarul Islam MD, Adrian O, Vladutiu MD, Theresa Donahue MT (2000) CD8 expression on B cells in chronic lymphocytic leukemia. Arch Pathol Lab Med 124:1361–1363Google Scholar
  33. 33.
    Giovanni Carulli MD, Alessandra S, Alessandra Marini MD (2009) Aberrant expression of CD8 in B-cell non-Hodgkin lymphoma. Am J Clin Pathol 132:186–190CrossRefPubMedGoogle Scholar
  34. 34.
    Morimura T, Ohashi K, Kon Y, Hattori M, Sugimoto C, Onuma M (1996) Apoptosis and CD8-down-regulation in the thymus of chickens infected with Marek’s disease virus. Arch Virol 141:2243–2249CrossRefPubMedGoogle Scholar
  35. 35.
    Kanehisa M, Goto S, Furumichi M, Tanabe M, Hirakawa M (2010) KEGG for representation and analysis of molecular networks involving diseases and drugs. Nucleic Acids Res 38:355–360CrossRefGoogle Scholar
  36. 36.
    Abdul Careem MF, Hunter BD (2007) Cytokine gene expression patterns associated with immunization against Marek’s disease in chickens. Vaccine 25:424–432CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Qi Xu
    • 1
  • Yang Chen
    • 1
  • Wen Ming Zhao
    • 1
  • Zheng Yang Huang
    • 1
  • Xiu Jun Duan
    • 2
  • Yi Yu Tong
    • 1
  • Yang Zhang
    • 1
  • Xiu Li
    • 1
  • Guo Bin Chang
    • 1
  • Guo Hong Chen
    • 1
    Email author
  1. 1.Key Laboratory of Animal Genetics and Breeding and Molecular Design of Jiangsu ProvinceYangzhou UniversityYangzhouPeople’s Republic of China
  2. 2.Chinese Waterfowl Germplasm Resource PoolTaizhouPeople’s Republic of China

Personalised recommendations