Antiviral activity of interferon-like cytokine (ILC) produced by head kidney leucocytes of common carp, Cyprinus carpio L

잉어 두신 백혈구에서 생성된 Interferon-like cytokine (ILC)의 항바이러스 활성

  • Published : 2004.08.30

Abstract

Interferons are kinds of cytokines that play an important role in antiviral defense mechanisms during viral infection by converting cells to synthesize proteins that inhibit viral replication. In the present study, an antiviral response against Spring viremia of carp virus (SVCV) was developed in common carp, Cyprinus carpio following stimulation with the potent interferon inducer, poly inosinic : cytidylic acid (poly I:C). Poly I:C injected fish showed lower cumulative mortalities against SVCV than untreated fish did. Moreover, in vitro study showed that head kidney leucocytes (HKLs) produced an interferon-like cytokine (ILC) after stimulation with poly I:C. According cytopathic effect reduction (CPER) assay to examine antiviral activity of crude ILC, the capacity of HKLs for ILC production was highest at 1×$10^6$cells/ml and significantly enhanced when treated with 20~50$\mu{g}$/ml of poly I:C. The optimal temperature and concentration of FBS to induce the highest ILC production were $20^\circ{C}$ and 5%, respectively.

인터페론은 바이러스에 감염된 세포에서 생성되어 다른 세포로 하여금 바이러스의 증식을 억제하는 단백질을 합성하게 하는 사이토카인의 일종으로서 바이러스에 대한 방어작용에서 중요한 역할을 담당한다. 본 연구 결과 강력한 인터페론 유도물질로 알려져 있는 poly inosinic : cytidylic acid (poly I:C)를 잉어에 주사한 결과 SVCV에 대한 항바이러스 활성을 확인할 수 있었다. 즉, Poly I:C 주사구에서 대조구에 비해 누적폐사율이 감소하였으며, 또한 두신백혈구를 분리하여, poly I:C를 처리한 결과 interferon-like cytokine (ILC)이 생성되었다. Crude ILC의 항바이러스 활성을 cytopathic effect reduction (CPER) assay로 조사한 결과, 적정 HKLs 농도는 1×$10^6$cells/ml으로 나타났으며, 20~50$\mu{g}$/ml 농도의 poly I:C를 처리했을 때 유의적으로 증가하였다. ILC 생성을 위한 적정온도 및 FBS의 농도는 각각 20$20^\circ{C}$와 5%로 나타났다.

Keywords

References

  1. Baudouy A. M. : Host-virus relationship during experimental spring viremia in carp. C. R. Acad. Sci. Hebd. Seances. Acad. Sci. D., 286 : 1225-1228, 1978
  2. Congleton, J. and Sun, B. : Interferon-like activity produced by anterior kidney leucocytes of rainbow trout stimulated in vitro by infectious hematopoietic necrosis virus or Poly I:C. Dis. Aquat. Org., 25 : 185-195, 1996 https://doi.org/10.3354/dao025185
  3. de Kinkelin P. and Dorson, M. : Interferon production in rainbow trout (Salmo gairdneri) experimentally infected with egtved virus. J. Gen. Virol., 19 : 125-127, 1973 https://doi.org/10.1099/0022-1317-19-1-125
  4. de Kinkelin, Dorson, P., M. and Hattenberger-Baudouy, A. M. : Interferon synthesis in trout and carp after viral infection. Dev. Comp. Immunol., Suppl., 2 : 167-174, 1982
  5. Eaton W. D. : Anti-viral activity in four species of salmonids following exposure to poly inosinic : cytidylic acid. Dis. Aquat. Org., 9 : 193-198, 1990 https://doi.org/10.3354/dao009193
  6. Fijan, N., Petrinec, Z., Sulimanovic, D. and Zwillenberg, L. O. : Isolation of viral causative agent from the acute form of infectious dropsy of carp. Vet. Arh., 41 : 125-138, 1971
  7. Giron, D. J., Smith, K. C., Hoffman, J. A. and Fowler, A. K. : Enhancement of viral RNA-induced interferon production in L cells treated with insulin and Amphotericin B methyl ester. J. Interferon Res., 1(4) : 581-586, 1981 https://doi.org/10.1089/jir.1981.1.581
  8. Graham, S. and Secombes, C. J. : Do fish lymphocytes secrete interferon-$\gamma$? J. Fish Biol., 36 : 563-573, 1990 https://doi.org/10.1111/j.1095-8649.1990.tb03557.x
  9. Lampson, G. P., Field, A. K., Tytell, A. A. and Hilleman, M. R. : Poly I:C/Poly-L-Lysin : potent inducer of interferons in promates. J. Interferon Res., 1(4) : 539-549, 1981 https://doi.org/10.1089/jir.1981.1.539
  10. Jensen, I and Robertsen, B. : Effects of double-stranded RNA and interferon on the antiviral activity of Atlantic salmon cells against infectious salmon anemia virus and infectious pancreatic necrosis virus. Fish & Shellfish Immunol., 13 : 221-241, 2002 https://doi.org/10.1006/fsim.2001.0397
  11. Jorgensen, J., Johansen, B. A., Stenersen, B. and Sommer, A. I. : CpG oligodeoxynucleotides and plasmid DNA stimulate Atlantic salmon (Salmo salar L.) leucocytes to produce supernatant with antiviral activity. Dev. Comp. Immunol., 25 : 313-321, 2001 https://doi.org/10.1016/S0145-305X(00)00068-9
  12. MacDonald, R. D. and Kennedy, J. C. : Infectious pancreatic necrosis virus persistently infects chinook salmon embryo cells independent of interferon. Virology, 95 : 260-264, 1979 https://doi.org/10.1016/0042-6822(79)90428-8
  13. Nygaard, R., Husgrad, S., Sommer, A. I., Leong, J. A. C. and Robertsen, B. : Induction of Mx protein by interferon and double-stranded RNA in salmonid cells. Fish & Shellfish Immunol., 10 : 435-450, 2000 https://doi.org/10.1006/fsim.1999.0249
  14. Renault, T., Torchy, C. and de kinkelin, P. : Spectrophotometric method for titration of trout interferon, and its application to rainbow trout fry experimentally infected with viral haemorrhagic septicaemia virus. Dis. Aquat. Org., 10 : 23-29, 1991 https://doi.org/10.3354/dao010023
  15. Secombes, C. J. : Interferon assessment. In Technique in fish immunology, pp. 71-77. Stolen, J. S., Fletcher, T. C., Rowley, A. F. Zelikoff, J. T., Kaattari, S. L. and Smith, S. A. eds., Fair Haven, NJ : SOS Publications, 1994