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Published in: European Journal of Medical Research 1/2013

Open Access 01-12-2013 | Research

Leflunomide inhibits the apoptosis of human embryonic lung fibroblasts infected by human cytomegalovirus

Authors: Ren Qi, Zeng Hua-Song, Zeng Xiao-Feng

Published in: European Journal of Medical Research | Issue 1/2013

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Abstract

Background

The immunomodulatory drug leflunomide (LEF) is frequently used for treating human cytomegalovirus (HCMV), but its antiviral mechanism is still unclear. In this study,we therefore investigated the effects of the active LEF metabolite A771726 on the HCMV lifecycle in human embryonic lung fibroblasts. We clarified the mechanism of LEF antiviral infection, and provide a new way to treat immune dysfunction patients with HCMV infection.

Methods

The experiment was divided into four groups: the control group, the HCMV group, the ganciclovir + HCMV group as well as the LEF + HCMV group. MTT was usedfor assessment of the cell inhibitory rate. Apoptosis was measured by staining with fluorescein isothiocyanate Annexin V and propidium iodide. Statistical significance was determined by paired t- test using SPSS software.

Results

The results of the study showed that cell proliferation was significantly inhibited by HCMV at 24 hours and 48 hours. With increasing HCMV concentration, the value-added inhibition of the cells was significantly decreased compared with the control group, and was statistically significant (P <0.01). Ganciclovir can increase proliferation of cellsinfected with HCMV; compared with the control group it was statistically significant (P <0.05). Meanwhile, with LEF treatment cell proliferation was significantly improved at 24 hours and 48 hours, with statistical significance (P <0.05). The apoptosis rate of human embryonic lung fibroblasts infected with HCMV increased significantly at 24 hours, 48 hours and 72 hours, and as time goes on the apoptosis rate increases statistically significantly (P <0.01) compared with the control group The apoptosis rate of theHCMV infection group decreased by adding LEF,and was statistically significant (P <0.05).

Conclusions

In this studywe show that LEF is an exciting new drug for cytomegalovirus infection. LEF significantly inhibited HCMV infection-induced apoptosis and proliferation, playing an important role in the treatment of patients infected by HCMV. In this study we explored the potential usefulness of LEF for cytomegalovirus infection and found it to be a cost-effective new treatment for cytomegalovirus infection that deserves further study.
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Literature
1.
go back to reference Jr Cinatal J, Schob M, Doerr HW: Role of tumor cell immune escape mechanisms in cytomegalovirus-mediated oncomodulation. Med Res Rev 2005, 25: 167–185. 10.1002/med.20018CrossRef Jr Cinatal J, Schob M, Doerr HW: Role of tumor cell immune escape mechanisms in cytomegalovirus-mediated oncomodulation. Med Res Rev 2005, 25: 167–185. 10.1002/med.20018CrossRef
2.
go back to reference Reuter JD: Cytomegalovirus induces T-cell independent apoptosis in brain during immune deficiency. J Clin Virol 2005, 32: 218–223. 10.1016/j.jcv.2004.07.012CrossRefPubMed Reuter JD: Cytomegalovirus induces T-cell independent apoptosis in brain during immune deficiency. J Clin Virol 2005, 32: 218–223. 10.1016/j.jcv.2004.07.012CrossRefPubMed
3.
go back to reference Goldmacher VS: Cell death suppression by cytomegaloviruses. Apoptosis 2005, 10: 251–265. 10.1007/s10495-005-0800-zCrossRefPubMed Goldmacher VS: Cell death suppression by cytomegaloviruses. Apoptosis 2005, 10: 251–265. 10.1007/s10495-005-0800-zCrossRefPubMed
4.
go back to reference Leger DY, Liagre B, Beneytout JL: Low dose leflunomide activates PI3K/Akt signalling in erythroleukemia cells and reduces apoptosis induced by anticancer agents. Apoptosis 2006, 11: 1747–1760. 10.1007/s10495-006-9439-7CrossRefPubMed Leger DY, Liagre B, Beneytout JL: Low dose leflunomide activates PI3K/Akt signalling in erythroleukemia cells and reduces apoptosis induced by anticancer agents. Apoptosis 2006, 11: 1747–1760. 10.1007/s10495-006-9439-7CrossRefPubMed
5.
go back to reference Josephson MA, Williams JW, Chandraker A, Randhawa PS: Polyomavirus-associated nephropathy: update on antiviral strategies. Transpl Infect Dis 2006, 8: 95–101. 10.1111/j.1399-3062.2006.00150.xCrossRefPubMed Josephson MA, Williams JW, Chandraker A, Randhawa PS: Polyomavirus-associated nephropathy: update on antiviral strategies. Transpl Infect Dis 2006, 8: 95–101. 10.1111/j.1399-3062.2006.00150.xCrossRefPubMed
6.
go back to reference Josephson MA, Jwaid B, Kadambi PV, Meehan SM, Williams JW: Leflunomide in solid organ transplantation and polyoma virus infection. Adv Exp Med Biol 2006, 577: 255–265. 10.1007/0-387-32957-9_18CrossRefPubMed Josephson MA, Jwaid B, Kadambi PV, Meehan SM, Williams JW: Leflunomide in solid organ transplantation and polyoma virus infection. Adv Exp Med Biol 2006, 577: 255–265. 10.1007/0-387-32957-9_18CrossRefPubMed
7.
go back to reference Ehlert K, Groll AH, Kuehn J, Vormoor J: Treatment of refractory CMV-infection following hematopoietic stem cell transplantation with the combination of fosmet and leflunomide. Klin Padiatr 2006, 218: 180–184. 10.1055/s-2006-933412CrossRefPubMed Ehlert K, Groll AH, Kuehn J, Vormoor J: Treatment of refractory CMV-infection following hematopoietic stem cell transplantation with the combination of fosmet and leflunomide. Klin Padiatr 2006, 218: 180–184. 10.1055/s-2006-933412CrossRefPubMed
8.
go back to reference Zeng H, Waldman WJ, Yin DP, Knight DA, Shen J, Ma L, Meister GT, Chong AS, Williams JW: Mechanistic study of malononitrileamide FK788 in cardiac transplantation and CMV infection in rats. Transplantation 2005, 79: 17–22. 10.1097/01.TP.0000137334.46155.94CrossRefPubMed Zeng H, Waldman WJ, Yin DP, Knight DA, Shen J, Ma L, Meister GT, Chong AS, Williams JW: Mechanistic study of malononitrileamide FK788 in cardiac transplantation and CMV infection in rats. Transplantation 2005, 79: 17–22. 10.1097/01.TP.0000137334.46155.94CrossRefPubMed
9.
go back to reference Koch S, Larbi A, Ozcelik D, Solana R, Gouttefangeas C, Attig S, Wikby A, Strindhall J, Franceschi C, Pawelec G: Cytomegalovirus infection: a driving force in human T cell immunosenescence. Ann N Y Acad Sci 2007, 1114: 23–35. 10.1196/annals.1396.043CrossRefPubMed Koch S, Larbi A, Ozcelik D, Solana R, Gouttefangeas C, Attig S, Wikby A, Strindhall J, Franceschi C, Pawelec G: Cytomegalovirus infection: a driving force in human T cell immunosenescence. Ann N Y Acad Sci 2007, 1114: 23–35. 10.1196/annals.1396.043CrossRefPubMed
10.
go back to reference Nakajima A, Yamanaka H, Kamatani N: Leflunomide: clinical effectiveness and mechanism of action. Clin Calcium 2003, 13: 771–775.PubMed Nakajima A, Yamanaka H, Kamatani N: Leflunomide: clinical effectiveness and mechanism of action. Clin Calcium 2003, 13: 771–775.PubMed
11.
go back to reference Levi ME, Mandava N, Chan LK, Weinberg A, Olson JL: Treatment of multidrug-resistant cytomegalovirus retinitis with systemically administered leflunomide. Transpl Infect Dis 2006, 8(1):38–43. 10.1111/j.1399-3062.2006.00128.xCrossRefPubMed Levi ME, Mandava N, Chan LK, Weinberg A, Olson JL: Treatment of multidrug-resistant cytomegalovirus retinitis with systemically administered leflunomide. Transpl Infect Dis 2006, 8(1):38–43. 10.1111/j.1399-3062.2006.00128.xCrossRefPubMed
12.
go back to reference Chong AS, Zeng H, Knight DA, Shen J, Meister GT, Williams JW, Waldman WJ: Concurrent antiviral and immunosuppressive activities of leflunomide invivo. Am J Transpl 2006, 6: 69–75. 10.1111/j.1600-6143.2005.01152.xCrossRef Chong AS, Zeng H, Knight DA, Shen J, Meister GT, Williams JW, Waldman WJ: Concurrent antiviral and immunosuppressive activities of leflunomide invivo. Am J Transpl 2006, 6: 69–75. 10.1111/j.1600-6143.2005.01152.xCrossRef
13.
go back to reference John GT, Manivannan J, Chandy S, Peter S, Fleming DH, Chandy SJ, Balakrishnan N, Krishnamurthy K, Kirubakaran MG, Jacob CK: A prospective evaluation of leflunomide therapy for cytomegalovirus disease in renal transplant recipients. Transplant Proc 2005, 37: 4303–4305. 10.1016/j.transproceed.2005.10.116CrossRefPubMed John GT, Manivannan J, Chandy S, Peter S, Fleming DH, Chandy SJ, Balakrishnan N, Krishnamurthy K, Kirubakaran MG, Jacob CK: A prospective evaluation of leflunomide therapy for cytomegalovirus disease in renal transplant recipients. Transplant Proc 2005, 37: 4303–4305. 10.1016/j.transproceed.2005.10.116CrossRefPubMed
14.
go back to reference Fitzsimmons WE, First MR: FK778, a synthetic malononitrilamide. Yonsei Med J 2004, 45: 1132–1135.CrossRefPubMed Fitzsimmons WE, First MR: FK778, a synthetic malononitrilamide. Yonsei Med J 2004, 45: 1132–1135.CrossRefPubMed
15.
go back to reference Avery RK, Bolwell BJ, Yen-Lieberman B, Lurain N, Waldman WJ, Longworth DL, Taege AJ, Mossad SB, Kohn D, Long JR, Curtis J, Kalaycio M, Pohlman B, Williams JW: Use of leflunomide in an allogeneic bone marrow transplant recipient with refractory cytomegalovirus infection. Bone Marrow Transplant 2004, 34: 1071–1975. 10.1038/sj.bmt.1704694CrossRefPubMed Avery RK, Bolwell BJ, Yen-Lieberman B, Lurain N, Waldman WJ, Longworth DL, Taege AJ, Mossad SB, Kohn D, Long JR, Curtis J, Kalaycio M, Pohlman B, Williams JW: Use of leflunomide in an allogeneic bone marrow transplant recipient with refractory cytomegalovirus infection. Bone Marrow Transplant 2004, 34: 1071–1975. 10.1038/sj.bmt.1704694CrossRefPubMed
16.
go back to reference Sarldlart-Direskeneli G, Inanc M, Fresko I, Akkoc N, Dalkilic E, Erken E, Karaaslan Y, Kinikli G, Oksel F, Pay S, Yucel E, Yentür SP, Duymaz-Tozkir J, Yilmaz V, Inanc N, Yazici H, Konice M, Direskeneli H: The role of HLA-DRB1 shared epitope alleles in predicting short-term response to leflunomide in rheumatoid arthritis. Rheumatology (Oxford) 2007, 46: 1842–1844. 10.1093/rheumatology/kem278CrossRef Sarldlart-Direskeneli G, Inanc M, Fresko I, Akkoc N, Dalkilic E, Erken E, Karaaslan Y, Kinikli G, Oksel F, Pay S, Yucel E, Yentür SP, Duymaz-Tozkir J, Yilmaz V, Inanc N, Yazici H, Konice M, Direskeneli H: The role of HLA-DRB1 shared epitope alleles in predicting short-term response to leflunomide in rheumatoid arthritis. Rheumatology (Oxford) 2007, 46: 1842–1844. 10.1093/rheumatology/kem278CrossRef
17.
go back to reference Sudamanam TD, Sahni RD, John GT: Leflunomide: a possible alternative for ganciclovir sensitive and resistant cytomegalovirus infections. Postgrad Med J 2006, 82: 313–314. 10.1136/pgmj.2005.038521CrossRef Sudamanam TD, Sahni RD, John GT: Leflunomide: a possible alternative for ganciclovir sensitive and resistant cytomegalovirus infections. Postgrad Med J 2006, 82: 313–314. 10.1136/pgmj.2005.038521CrossRef
18.
go back to reference Snydman DR: Leflunomide: a small step forward in meeting the urgent need for treatment of drug-resistant cytomegalovirus infection. Transplantation 2010, 90: 362–363.CrossRefPubMed Snydman DR: Leflunomide: a small step forward in meeting the urgent need for treatment of drug-resistant cytomegalovirus infection. Transplantation 2010, 90: 362–363.CrossRefPubMed
19.
go back to reference Teschner S, Burst V: Leflunomide: a drug with a potential beyond rheumatology. Immunotherapy 2010, 2: 637–650. 10.2217/imt.10.52CrossRefPubMed Teschner S, Burst V: Leflunomide: a drug with a potential beyond rheumatology. Immunotherapy 2010, 2: 637–650. 10.2217/imt.10.52CrossRefPubMed
20.
go back to reference Eid AJ, Razonable RR: New developments in the management of cytomegalovirus infection after solid organ transplantation. Drugs 2010, 70: 965–981. 10.2165/10898540-000000000-00000CrossRefPubMed Eid AJ, Razonable RR: New developments in the management of cytomegalovirus infection after solid organ transplantation. Drugs 2010, 70: 965–981. 10.2165/10898540-000000000-00000CrossRefPubMed
21.
go back to reference Webel R, Milbradt J, Auerochs S, Schregel V, Held C, Nöbauer K, Razzazi-Fazeli E, Jardin C, Wittenberg T, Sticht H, Marschall M: Two isoforms of the protein kinase pUL97 of human cytomegalovirus are differentially regulated in their nuclear translocation. J Gen Virol 2011, 92(3):638–649. 10.1099/vir.0.026799-0CrossRefPubMed Webel R, Milbradt J, Auerochs S, Schregel V, Held C, Nöbauer K, Razzazi-Fazeli E, Jardin C, Wittenberg T, Sticht H, Marschall M: Two isoforms of the protein kinase pUL97 of human cytomegalovirus are differentially regulated in their nuclear translocation. J Gen Virol 2011, 92(3):638–649. 10.1099/vir.0.026799-0CrossRefPubMed
Metadata
Title
Leflunomide inhibits the apoptosis of human embryonic lung fibroblasts infected by human cytomegalovirus
Authors
Ren Qi
Zeng Hua-Song
Zeng Xiao-Feng
Publication date
01-12-2013
Publisher
BioMed Central
Published in
European Journal of Medical Research / Issue 1/2013
Electronic ISSN: 2047-783X
DOI
https://doi.org/10.1186/2047-783X-18-3

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