Archives of Virology

, Volume 161, Issue 4, pp 913–928 | Cite as

A lack of Fas/FasL signalling leads to disturbances in the antiviral response during ectromelia virus infection

  • K. Bień
  • Z. Sobańska
  • J. Sokołowska
  • P. Bąska
  • Z. Nowak
  • A. Winnicka
  • M. Krzyzowska
Original Article


Ectromelia virus (ECTV) is an orthopoxvirus (OPV) that causes mousepox, the murine equivalent of human smallpox. Fas receptor-Fas ligand (FasL) signaling is involved in apoptosis of immune cells and virus-specific cytotoxicity. The Fas/FasL pathway also plays an important role in controlling the local inflammatory response during ECTV infection. Here, the immune response to the ECTV Moscow strain was examined in Fas (-) (lpr), FasL (-) (gld) and C57BL6 wild-type mice. During ECTV-MOS infection, Fas- and FasL mice showed increased viral titers, decreased total numbers of NK cells, CD4+ and CD8+ T cells followed by decreased percentages of IFN-γ expressing NK cells, CD4+ and CD8+ T cells in spleens and lymph nodes. At day 7 of ECTV-MOS infection, Fas- and FasL-deficient mice had the highest regulatory T cell (Treg) counts in spleen and lymph nodes in contrast to wild-type mice. Furthermore, at days 7 and 10 of the infection, we observed significantly higher numbers of PD-L1-expressing dendritic cells in Fas (-) and FasL (-) mice in comparison to wild-type mice. Experiments in co-cultures of CD4+ T cells and bone-marrow-derived dendritic cells showed that the lack of bilateral Fas-FasL signalling led to expansion of Tregs. In conclusion, our results demonstrate that during ECTV infection, Fas/FasL can regulate development of tolerogenic DCs and Tregs, leading to an ineffective immune response.


Dendritic Cell Natural Killer Cell West Nile Virus Antiviral Response FACSCalibur Flow Cytometer 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This work was the subject of two Master Thesis projects (for KB and ZS) and was co-funded by grants from the Department of Preclinical Sciences, Faculty of Veterinary Medicine, Warsaw University of Life Sciences.

Compliance with ethical standards

Conflict of interest

The authors declare that there is no conflict of interests regarding the publication of this paper.


  1. 1.
    Stanford MM, McFadden G, Karupiah G, Chaudhri G (2007) Immunopathogenesis of poxvirus infections: forecasting the impending storm. Immunol Cell Biol 85:93–102CrossRefPubMedGoogle Scholar
  2. 2.
    Esteban DJR, Buller ML (2005) Ectromelia virus: the causative agent of mousepox. J Gen Virol 861:2645–2659CrossRefGoogle Scholar
  3. 3.
    Wallace GD, Buller RM (1985) Kinetics of ectromelia virus (mousepox) transmission and clinical response in C57BL/6j, BALB/cByj and AKR/J inbred mice. Lab Anim Sci 35:41–46PubMedGoogle Scholar
  4. 4.
    Jacoby RO, Bhatt PN, Brownstein DG (1989) Evidence that NK cells and interferon are required for genetic resistance to lethal infection with ectromelia virus. Arch Virol 108:49–58CrossRefPubMedGoogle Scholar
  5. 5.
    Delano ML, Brownstein DG (1995) Innate resistance to lethal mousepox is genetically linked to the NK gene complex on chromosome 6 and correlates with early restriction of virus replication by cells with an NK phenotype. J Virol 69:5875–5877PubMedPubMedCentralGoogle Scholar
  6. 6.
    Ramaswamy M, Cleland SY, Cruz AC, Siegel RM (2009) Many checkpoints on the road to cell death: regulation of Fas-FasL interactions and Fas signaling in peripheral immune responses. Results Probl Cell Differ 49:17–47CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Davidson WF, Haudenschild C, Kwon J, Williams MS (2002) T cell receptor ligation triggers novel nonapoptotic cell death pathways that are Fas-independent or Fas-dependent. J Immunol 169:6218–6230CrossRefPubMedGoogle Scholar
  8. 8.
    Ju ST, Panka DJ, Cui H, Ettinger R et al (1995) Fas (CD95)/FasL interactions required for programmed cell death after T-cell activation. Nature 373:444–448CrossRefPubMedGoogle Scholar
  9. 9.
    Stranges PB, Watson J, Cooper CJ et al (2007) Elimination of antigen-presenting cells and autoreactive T cells by fas contributes to prevention of autoimmunity. Immunity 26:629–641CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Krzyzowska M, Baska P, Orlowski P et al (2013) HSV-2 regulates monocyte inflammatory response via the Fas/FasL pathway. PLoS One 8:e70308CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Lyons C, Fernandes P, Fanning LJ, Houston A, Brint E (2015) Engagement of Fas on macrophages modulates poly I:C induced cytokine production with specific enhancement of IP-10. PLoS One 10:e0123635CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Shrestha B, Diamond MS (2007) Fas ligand interactions contribute to CD8 T-cell-mediated control of West Nile virus infection in the central nervous system. J Virol 81:11749–11757CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Johnson J, Chu CF, Milligan GN (2008) Effector CD4 T-cell involvement in clearance of infectious herpes simplex virus type 1 from sensory ganglia and spinal cords. J Virol 82:9678–9688CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Topham DJ, Tripp RA, Doherty PC (1997) CD8 T cells clear influenza virus by perforin or Fas-dependent processes. J Immunol 159:5197–5200PubMedGoogle Scholar
  15. 15.
    Parra B, Lin MT, Stohlman SA et al (2000) Contributions of Fas-Fas ligand interactions to the pathogenesis of mouse hepatitis virus in the central nervous system. J Virol 74:2447–2450CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Chaudhri G, Panchanathan V, Buller RM et al (2004) Polarized type 1 cytokine response and cell-mediated immunity determine genetic resistance to mousepox. Proc Natl Acad Sci USA 101:9057–9062CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Trapani A, Jans DA, Jans PJ et al (1998) Efficient nuclear targeting of granzyme B and the nu- clear consequences of apoptosis induced by granzyme B and perforin are caspase-dependent, but cell death is caspase-independent. J Biol Chem 273:27934–27938CrossRefPubMedGoogle Scholar
  18. 18.
    Turner SJ, Silke J, Kenshole B, Ruby J (2000) Characterization of the ectromelia virus serpin, SPI-2. J Gen Virol 81:2425–2430CrossRefPubMedGoogle Scholar
  19. 19.
    Krzyzowska M, Polanczyk M, Bas M et al (2005) Mousepox conjunctivitis: the role of Fas/FasL-mediated apoptosis of epithelial cells in virus dissemination. J Gen Virol 86:2007–2018CrossRefPubMedGoogle Scholar
  20. 20.
    Krzyzowska M, Cymerys J, Winnicka A, Niemiałtowski M (2006) Involvement of Fas and FasL in Ectromelia virus-induced apoptosis in mouse brain. Virus Res 115:141–149CrossRefPubMedGoogle Scholar
  21. 21.
    Bień K, Sokołowska J, Bąska P et al (2015) Fas/FasL pathway participates in regulation of antiviral and inflammatory response during mousepox infection of lungs. Mediators Inflamm 2015:281613. doi: 10.1155/2015/281613 PubMedPubMedCentralGoogle Scholar
  22. 22.
    Krzyzowska M, Orłowski P, Bąska P, Bodera P, Zdanowski R, Stankiewicz W (2014) Role of Fas/FasL signaling in regulation of anti-viral response during HSV-2 vaginal infection in mice. Immunobiology 219:932–943CrossRefPubMedGoogle Scholar
  23. 23.
    Gunalp A (1965) Growth and cytopathic effect of rubella virus in a line of green monkey kidney cells. Proc Soc Exp Biol Med 118:185–190CrossRefGoogle Scholar
  24. 24.
    Krzyzowska M, Schollenberger A, Skierski J, Niemialtowski M (2002) Apoptosis during ectromelia orthopoxvirus infection is DEVDase dependent: in vitro and in vivo studies. Microbes Infect 4:599–611CrossRefPubMedGoogle Scholar
  25. 25.
    Amoah S, Holbrook BC, Yammani RD, Alexander-Miller MA (2013) High viral burden restricts short-lived effector cell number at late times postinfection through increased natural regulatory T cell expansion. J Immunol 190:5020–5029CrossRefPubMedPubMedCentralGoogle Scholar
  26. 26.
    Strasser A, Jost PJ, Nagata S (2009) The many roles of FAS receptor signaling in the immune system. Immunity 30:180–192CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Müllbacher A, Wallich R, Moyer RW, Simon MM (1999) Poxvirus-encoded serpins do not prevent cytolytic T cell-mediated recovery from primary infections. J Immunol 162:7315–7321PubMedGoogle Scholar
  28. 28.
    Brownstein DG, Gras L (1995) Chromosome mapping of Rmp-4, a gonad-dependent gene encoding host resistance to mousepox. J Virol 69:6958–6964PubMedPubMedCentralGoogle Scholar
  29. 29.
    Biron A, Nguyen KB, Pien GC, Cousens LP, Salazar-Mather TP (1999) Natural killer cells in antiviral defense: function and regulation by innate cytokines. Annu Rev Immunol 17:189–220CrossRefPubMedGoogle Scholar
  30. 30.
    Parker K, Parker S, Yokoyama WM, Corbett JA, Buller RM (2007) Induction of natural killer cell responses by ectromelia virus controls infection. J Virol 81:4070–4079CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Fang LL, Lanier L, Sigal J (2008) A role for NKG2D in NK cell-mediated resistance to poxvirus disease. PLoS Pathog 4:e30CrossRefPubMedPubMedCentralGoogle Scholar
  32. 32.
    Meyer H, Damon K, Esposito JJ (2004) Orthopoxvirus diagnostics. Methods Mol Biol 269:119–133PubMedGoogle Scholar
  33. 33.
    Karupiah G, Buller RM, Rooijen Van et al (1996) Different roles for CD4+ and CD8+ T lymphocytes and macrophage subsets in the control of a generalized virus infection. J Virol 70:8301–8309PubMedPubMedCentralGoogle Scholar
  34. 34.
    Fang M, Siciliano NA, Hersperger AR et al (2012) Perforin-dependent CD4+ T-cell cytotoxicity contributes to control a murine poxvirus infection. Proc Natl Acad Sci USA 109:9983–9988CrossRefPubMedPubMedCentralGoogle Scholar
  35. 35.
    Seedhom MO, Keisha S, Mathurin KS, Kim SK, Welsh RM (2012) Increased protection from Vaccinia Virus infection in mice genetically prone to lymphoproliferative disorders. J Virol 86:6010–6022CrossRefPubMedPubMedCentralGoogle Scholar
  36. 36.
    Josefowicz SZ, Lu LF, Rudensky AY (2012) Regulatory T cells: mechanisms of differentiation and function. Annu Rev Immunol 30:531–564CrossRefPubMedGoogle Scholar
  37. 37.
    Veiga-Parga T, Sehrawat S, Rouse BT (2013) Role of regulatory T cells during virus infection. Immunol Rev 255:182–196CrossRefPubMedPubMedCentralGoogle Scholar
  38. 38.
    Haeryfar SM, DiPaolo RJ, Tscharke DC, Bennink JR, Yewdell JW (2005) Regulatory T cells suppress CD8+ T cell responses induced by direct priming and cross-priming and moderate immunodominance disparities. J Immunol 174:3344–3351CrossRefPubMedGoogle Scholar
  39. 39.
    Weiss L, Donkova-Petrini V, Caccavelli L, Balbo M, Carbonneil C, Levy Y (2004) Human immunodeficiency virus-driven expansion of CD4+CD25+ regulatory T cells, which suppress HIV-specific CD4 T-cell responses in HIV-infected patients. Blood 104:3249–3256CrossRefPubMedGoogle Scholar
  40. 40.
    Nan XP, Zhang Y, Yu HT et al (2012) Inhibition of viral replication down-regulates CD4(+)CD25(high) regulatory T cells and programmed death-ligand 1 in chronic hepatitis B. Viral Immunol 25:21–28PubMedPubMedCentralGoogle Scholar
  41. 41.
    Ding Y, Xu J, Bromberg JS (2012) Regulatory T cell migration during an immune response. Trends Immunol 33:174–180CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Pletinckx K, Lutz MB (2014) Dendritic cells generated with Flt3L and exposed o apoptotic cells lack induction of T cell anergy and Foxp3+ regulatory T cell conversion in vitro. Immunobiology 219:230–240CrossRefPubMedGoogle Scholar
  43. 43.
    Engelmayer J, Larsson M, Subklewe M et al (1999) Vaccinia virus inhibits the maturation of human dendritic cells: a novel mechanism of immune evasion. J Immunol 163:6762–6768PubMedGoogle Scholar
  44. 44.
    Chatzigeorgiou A, Lyberi M, Chatzilymperis G et al (2009) CD40/CD40L signaling and its implication in health and disease. Biofactors 35:474–483CrossRefPubMedGoogle Scholar
  45. 45.
    Yao S, Chen L (2014) PD-1 as an immune modulatory receptor. Cancer J 20:262–264CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Kamphorst AO, Ahmed R (2013) Manipulating the PD-1 pathway to improve immunity. Curr Opin Immunol 25:381–388CrossRefPubMedGoogle Scholar
  47. 47.
    Wang X, Zhang Z, Zhang S et al (2008) B7–H1 up-regulation impairs myeloid DC and correlates with disease progression in chronic HIV-1 infection. Eur J Immunol 38:3226–3236CrossRefPubMedGoogle Scholar
  48. 48.
    Shen T, Chen X, Chen Y et al (2010) Increased PD-L1 expression and PD-L1/CD86 ratio on dendritic cells were associated with impaired dendritic cells function in HCV infection. J Med Virol 82:1152–1159CrossRefPubMedGoogle Scholar
  49. 49.
    Chentoufi AA, Dervillez X, Dasgupta G et al (2012) The herpes simplex virus type 1 latency-associated transcript inhibits phenotypic and functional maturation of dendritic cells. Viral Immunol 25:204–215PubMedPubMedCentralGoogle Scholar
  50. 50.
    Fritzsching B, Oberle N, Eberhardt N et al (2005) In contrast to effector T cells, CD4+CD25+FoxP3+ regulatory T cells are highly susceptible to CD95 ligand- but not to TCR-mediated cell death. J Immunol. 175:32–36CrossRefPubMedGoogle Scholar

Copyright information

© Springer-Verlag Wien 2016

Authors and Affiliations

  • K. Bień
    • 1
  • Z. Sobańska
    • 2
  • J. Sokołowska
    • 3
  • P. Bąska
    • 4
  • Z. Nowak
    • 5
  • A. Winnicka
    • 6
  • M. Krzyzowska
    • 1
  1. 1.Department of Regenerative MedicineMilitary Institute of Hygiene and EpidemiologyWarsawPoland
  2. 2.Department of Molecular Genetics, Faculty of Biology and Environmental ProtectionUniversity of LodzLodzPoland
  3. 3.Department of Morphological Sciences, Faculty of Veterinary MedicineWarsaw University of Life SciencesWarsawPoland
  4. 4.Department of Preclinical Sciences, Faculty of Veterinary MedicineWarsaw University of Life SciencesWarsawPoland
  5. 5.Department of Genetics and Animal Breeding, Faculty of Animal ScienceWarsaw University of Life SciencesWarsawPoland
  6. 6.Department of Pathology and Veterinary Diagnostics, Faculty of Veterinary MedicineWarsaw University of Life SciencesWarsawPoland

Personalised recommendations