MECHANISMS OF CELL RESISTANCE TO CYTOMEGALOVIRUS ARE CONNECTED WITH CELL PROLIFERATION STATE AND TRANSCRIPTION ACTIVITY OF LEUKOCYTE AND IMMUNE INTERFERON GENES

Abstract. Cytomegalovirus (CMV) infection in diploid human fibroblasts (HF) and levels of cell resistance to this virus were shown to be in direct correlation with high α-interferon (IFNα) gene activity and induction of IFNγ gene transcription. Regulation of IFNα mRNA transcription was revealed to be positively associated with cellular DNA synthesis. At the same time, activities of IFNβ and IFNγ genes were at the constantly low level and were not induced in DNA-synthetic phase (S-phase) of the cells. Levels of IFNα mRNA synthesis are quite different for G0- vs S-phase-synchronized HF110044 cell cultures: appropriate values for dividing cells (S-phase) proved to be 100-fold higher than in resting state (G0). The mode of CMV infection in resting HF-cell could be considered either as acute, or a productive one. On the contrary, proliferating cells exhibited lagging viral syntheses and delayed cell death. Arrest of CMV replication may be, to some extent, comparable with latent infectious state, being associated with high production of IFNα. Both basal and induced levels of IFNα mRNA in CMV-resistant adult human skin fibroblast cells (HSF-1608) were 10-fold higher than in human embryo lung cell line (HELF-977), which is highly sensitive to CMV. Moreover, a short-time induction of IFNγ genes was observed in resistant cells, whereas no such effect was noticed in highly sensitive cells. CMV reproduction in sensitive cell lines (HELF-977 and HELF-110044) partially inhibits IFNα mRNA transcription at the later stages of infection (24 to 48 hours). Thus, cellular resistance and control of CMV infection in diploid fibroblasts are associated predominantly with high transcription of IFNα gene, and with temporal induction of IFNγ gene. We did not reveal any participation of IFNβ genes in protection of human diploid fibroblasts from CMV.

human fibroblasts, interferon, mRNA transcription, cytomegalovirus infection

1. Вирусология. Методы / Под ред. Мейхи Б. – М.: Мир, 1988. – 290 с.

2. Ершов Ф.И., Киселев О.И. Интерфероны и их индукторы – М.: ГЭОТАР-МЕДИА, 2005. – 368 с.

3. Соколова Т.М., Бибикова О.В., Быстров Н.С., Урываев Л.В. Экспрессия генов системы интерферона и клеточного апоптоза в пробах крови человека // Вопр. вирусол. – 2005. – № 1. – С. 19-23.

4. Соколова Т.М., Урываев Л.В. Способ определения цитокинового статуса человека на генетическом уровне. – Пат. на изобретение № 2181177 от 27.04.2002, РФ.

5. Соколова Т.М., Федорова Н.Е., Меджидова М.Г., Терехов С.М., Урываев Л.В., Кущ А.А. Регуляция уровня генной активности 2`, 5`-олигоаденилатсинтетазы в фибробластах человека при цитомегаловирусной инфекции // Вопр. вирусол. – 2007. – № 1. – С. 28-36.

6. Федорова Н.Е., Меджидова А.А., Меджидова М.Г., Кущ А.А. Блок клеточной пролиферации и патологии митоза в клетках, инфицированных цитомегаловирусом: роль периода клеточного цикла в момент заражения // Докл. АН. – 2003. – Т. 392. – № 4. – С. 552-555.

7. Abate D.A., Watanabe S., Mocarski E.S. Major human cytomegalovirus structural protein pp65 (ppUL83) prevents interferon response factor 3 activation in the interferon response // J. Virol. – 2004. – Vol. 78. – P. 10995-11006.

8. Browne E., Wing B., Coleman D., Shenk T. Altered cellular mRNA levels in human cytomegalovirusinfected fibroblast: viral block to the accumulation of antiviral mRNAs // J. Virol. – 2001. – Vol. 75. – P. 12319-12330.

9. Browne E.P., Shenk T. Human cytomegalovirus UL83-coded pp65 virion protein inhibits antiviral gene expression in infected cells // Proc. Nat. Acad. Sci. USA. – 2003. – Vol. 100. – P. 11439-11444.

10. Challcombe J.F., Rechtsteiner A., Gottardo R., Rocha L.M., Browne E.P., Shenk T., Altherr M.R., Brettin T.S. Evaluation of the host transcriptional response to human cytomegalovirus infection // Physiol. Genomics. – 2004. – Vol. 18. – P. 51-62.

11. Chen J., Baig E., Fish E.N. Diversity and relatedness among type I interferons // J. Interferon Cytokine Res. – 2004. – Vol. 24. – P. 687-698.

12. Chomzynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thioocyanatephenol-chloroform extraction // Anal. Biochem. – 1987. – Vol. 162. – P. 156-159.

13. Dallman M.J., Porter A.C.G. Semiquantitative PCR for the analysis of gene expression // PCR. A practical approach / Ed. by M.J. McPherson et al. – Oxford, 1993. – P. 215-224.

14. Diaz M.O., Bohlander S., Allen G. Nomenclature of the human interferon genes // J. Interferon. Cytokine Res. –1996. – Vol. 16. – P. 179-180.

15. Goswami B.B., Kulka M., Ngo D., Cebula T.A. Apoptosis induced by a cytopathic hepatitis A virus is dependent on caspase activation following ribosomal RNA degradation but occurs in the absence of 2’-5’ oligoadenylate synthetase // Antiviral Res. – 2004. – Vol. 63. – P. 153-166.

16. Hakki M., Geballe A.P. Double-standed RNA binding by human cytomegalovirus pTRSI // J. Virol. – 2005. – Vol. 79. – P. 7311-7318.

17. Hertel L., Mocarski E.S. Global analysis of host cell gene expression late during cytomegalovirus infection reveals extensive dysregulation of cell cycle gene expression and induction of pseudomitosis independent of US28 function // J. Virol. – 2004. – Vol. 78. – P. 11988-12011.

18. Iyer V.R., Eisen M.B., Ross D.T., Schuler G., Moore T., Lee J.C., Trent J.M., Staudt L.M., Hudson J.Jr.,Boguski M.S., Lashkari D., Shalon D., Botstein D., Brown P.O. The transcriptional program in the response of human fibroblasts to serum // Science. – 1999. – Vol. 283. – P. 83-87.

19. Karpov A.V. Endogenous and exogenous interferons in HIV-infection // Eur. J. Med. Res. – 2001. – Vol. 6. – P. 507-524.

20. Knipe D.M., Samuel C.E., Palese P. Virushost cell interactions // Fields virology / Ed. By D.M. Knipe et al. – Lippincott-Raven, Philadelphia, 2001. – P. 133-170.

21. Kotenko S.V., Saccani S., Izotova L.S., Microchnitchenko O.V., Pestka S. Human cytomegalovirus harbors its own unique IL-10 homolog (cmv IL-10) // Proc. Nat. Acad. Sci. USA. – 2000. – Vol. 97. – P. 1695-1700.

22. Lee G.C., Yi H.A., Lee C.H. Stimulation of interferon-beta gene expression by human cytomegalovirus via nuclear factor kappa B and phosphatidylinositol 3-kinase pathway // Virus Res. – 2006. – Vol. 117. – P. 209-214.

23. Loenen W., Bruggemen C., Wiertz E. Immune evasion by human cytomegalovirus: lesions in immunology and cell biology // Semin. Immunol. – 2001. – Vol. 13. – P. 41-49.

24. Miller D.M., Rahill B.M., Boss J.M. Human cytomegalovirus inhibits major Histocompatibility complex class II expression by disruption of the JAK/STAT pathway // J. Exp. Med. – 1998. – Vol. 187. – P. 675-683.

25. Ohyama K., Sano T., Toyoda H. Predominant contribution of IFN-beta expression to apoptosis induction in human uterine cervical fibroblast cells by influenza-virus infection // Biol. Pharm. Bull. – 2004. – Vol. 27. – P. 1750-1757.

26. Paulus Ch., Krauss S., Nevels M. A human cytomegalovirus antagonist of type I IFN-dependent signal transducer and activator of transcription signaling // Proc. Natl. Acad. Sci. USA. – 2006. – Vol. 103. – P. 3840-3845.

27. Samuel C.E. Antiviral actions of interferons // Clin. Microb. Rev. – 2001. – Vol. 14. – P. 778-809.

28. Silvia O.J., Pantelic L., Mackenzie J.S., Shellam G.R., Paradimitriou J., Urosevic N. Virus spread, tissue inflammation and antiviral response in brains of flavivirus susceptible and resistant mice acutely infected with Murray Valley encephalitis virus // Arch. Virol. – 2004. – Vol. 149. – P. 447-464.

29. Simmen K., Singh J., Luukkonen B., Lopper M., Bittner A., Miller N., Jackson M., Compton T., Fruh K. Global modification of cellular transcription by human cytomegalovirus is initiated by viral glicoprotein B // Proc. Natl. Acad. Sci. USA. – 2001. – Vol. 98. – P. 7140-7145.

30. Spenser J.V., Lockridge K.M., Barry P.A., Lin G., Tsang M., Penfold M.E., Shall T.J. Potent immunosuppressive activities of cytomegalovirusencoded interleukin-10 // J. Virol. – 2002. – Vol. 76. – P. 1285-1292.

31. Strausberg R.L., Feingold E.A., Grouse L.H., Derge J.G., Klausner R.D., Collins F.S., Wagner L., Shenmen C.M., Schuler G.D., Altschul S.F., Zeeberg B., Buetow K.H., Schaefer C.F., Bhat N.K., Hopkins R.F., Jordan H., Moore T., Max S.I., Wang J., Hsieh F., Diatchenko L., Marusina K., Farmer A.A., Rubin G.M., Hong L., Stapleton M., Soares M.B., Bonaldo M.F., Casavant T.L., Scheetz T.E., Brownstein M.J., Usdin T.B., Toshiyuki S., Carninci P., Prange C., Raha S.S., Loquellano N.A., Peters G.J., Abramson R.D., Mullahy S.J., Bosak S.A., McEwan P.J., McKernan K.J., Malek J.A., Gunaratne P.H., Richards S., Worley K.C., Hale S., Garcia A.M., Gay L.J., Hulyk S.W., Villalon D.K., Muzny D.M., Sodergren E.J., Lu X., Gibbs R.A., Fahey J., Helton E., Ketteman M., Madan A., Rodrigues S., Sanchez A., Whiting M., Madan A., Young A.C., Shevchenko Y., Bouffard G.G., Blakesley R.W., Touchman J.W., Green E.D., Dickson M.C., Rodriguez A.C., Grimwood J., Schmutz J., Myers R.M., Butterfield Y.S.N., Krzywinski M.I., Skalska U., Smailus D.E., Schnerch A., Schein J.E., Jones S.J.M., Marra M.A. Generation and initial analysis of more than 15,000 full-length human and mouse cDNA sequences // Proc. Nat. Acad. Sci. USA. – 2002. – Vol. 99. – P. 16899-16903.

32. Taylor R.T., Bresnahan W.A. Human cytomegalovirus immediate-early 2 gene expression blocks virus-induced beta interferon production // J. Virol. – 2005. – Vol. 79. – P. 3873-3877.

33. Zhu H., Cong J.P., Mamtora J., Gingeras T., Shenk T. Cellular gene expression altered by human cytomegalovirus: global monitoring with oligonucleotide arrays // Proc. Nat. Acad. Sci. USA. – 1998. – Vol. 95. – P. 14470-14475.

34. Zimmerman A., Trilling M., Wagner M., Wilborn M., Bubic I., Jonjic S., Koszinowski U., Hengel H. A cytomegaloviral protein reveals a dual role for STAT2 in IFNγ signaling and antiviral responses // J. Exp.Med. – 2005. – Vol. 201. – P. 1543-1553.