Vitamin E inhibits cyclosporin A and H2O2 promoted Epstein-Barr virus (EBV) transformation of human B cells as assayed by EBV oncogene LMP1 expression

Changguo Chen, Kunam Sudhakar Reddy, T. D. Johnston, T. T. Khan, D. Ranjan

Research output: Contribution to journalArticle

6 Citations (Scopus)

Abstract

Background. We have previously shown that oxidative stress induced by H2O2 or cyclosporin A (CsA) can promote Epstein-Barr virus (EBV) transformation of human B cells as analyzed by colony formation, cell number, and by 3H-thymidine incorporation. In this report, we used EBV oncogene LMP1 as a marker to analyze H2O2 or CsA promotion of EBV transformation of human B cells and to test whether antioxidant vitamin E could inhibit H2O2 or CsA promoted LMP1 expression in the EBV-infected cells. Materials and methods. Human splenocytes were prepared by centrifugation and plating technique to provide a greater than 80% pure preparation of B cells and were used for EBV infection. The EBV infected cells were treated with H2O2 (0.1 mM, 10 min), or with CsA (500 ng/ml) with or with out vitamin E (40 μM). The cells were cultured for up to 4 weeks. Samples were taken every week and were stained with phycoerythrin-conjugated mouse anti-LMP1 monoclonal antibody to assay LMP1 positive population by flow cytometry. Results. In EBV-infected cells, the LMP1-positive cell population reached 14% after 4 weeks of culture. CsA or H2O2 treatment promoted LMP1 positive population to 43% and 41% after 4 weeks of culture. Vitamin E (40 μM) completely inhibited LMP1 expression in EBV-infected cells and in CsA- or H2O 2-treated cells. Conclusion. In agreement with our previous observation, CsA or H2O2 can promote EBV transformation of human B cells. This oxidative stress induced promotion of EBV transformation can be blocked by antioxidant Vitamin E. This finding may have future therapeutic implications for post-transplant lymphoproliferative disorder.

Original languageEnglish (US)
Pages (from-to)228-233
Number of pages6
JournalJournal of Surgical Research
Volume113
Issue number2
DOIs
StatePublished - Aug 2003
Externally publishedYes

Fingerprint

Human Herpesvirus 4
Vitamin E
Oncogenes
Cyclosporine
B-Lymphocytes
Oxidative Stress
Antioxidants
Phycoerythrin
Population
Epstein-Barr Virus Infections
Lymphoproliferative Disorders
Centrifugation
Thymidine
Cultured Cells
Flow Cytometry
Cell Count
Monoclonal Antibodies
Observation
Transplants

Keywords

  • Cyclosporin A
  • Epstein-Barr virus
  • HO
  • Oxidative stress
  • Post-transplant lymphoproliferative disorders (PTLD)
  • Vitamin E (Vit. E)

ASJC Scopus subject areas

  • Surgery

Cite this

Vitamin E inhibits cyclosporin A and H2O2 promoted Epstein-Barr virus (EBV) transformation of human B cells as assayed by EBV oncogene LMP1 expression. / Chen, Changguo; Reddy, Kunam Sudhakar; Johnston, T. D.; Khan, T. T.; Ranjan, D.

In: Journal of Surgical Research, Vol. 113, No. 2, 08.2003, p. 228-233.

Research output: Contribution to journalArticle

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title = "Vitamin E inhibits cyclosporin A and H2O2 promoted Epstein-Barr virus (EBV) transformation of human B cells as assayed by EBV oncogene LMP1 expression",
abstract = "Background. We have previously shown that oxidative stress induced by H2O2 or cyclosporin A (CsA) can promote Epstein-Barr virus (EBV) transformation of human B cells as analyzed by colony formation, cell number, and by 3H-thymidine incorporation. In this report, we used EBV oncogene LMP1 as a marker to analyze H2O2 or CsA promotion of EBV transformation of human B cells and to test whether antioxidant vitamin E could inhibit H2O2 or CsA promoted LMP1 expression in the EBV-infected cells. Materials and methods. Human splenocytes were prepared by centrifugation and plating technique to provide a greater than 80{\%} pure preparation of B cells and were used for EBV infection. The EBV infected cells were treated with H2O2 (0.1 mM, 10 min), or with CsA (500 ng/ml) with or with out vitamin E (40 μM). The cells were cultured for up to 4 weeks. Samples were taken every week and were stained with phycoerythrin-conjugated mouse anti-LMP1 monoclonal antibody to assay LMP1 positive population by flow cytometry. Results. In EBV-infected cells, the LMP1-positive cell population reached 14{\%} after 4 weeks of culture. CsA or H2O2 treatment promoted LMP1 positive population to 43{\%} and 41{\%} after 4 weeks of culture. Vitamin E (40 μM) completely inhibited LMP1 expression in EBV-infected cells and in CsA- or H2O 2-treated cells. Conclusion. In agreement with our previous observation, CsA or H2O2 can promote EBV transformation of human B cells. This oxidative stress induced promotion of EBV transformation can be blocked by antioxidant Vitamin E. This finding may have future therapeutic implications for post-transplant lymphoproliferative disorder.",
keywords = "Cyclosporin A, Epstein-Barr virus, HO, Oxidative stress, Post-transplant lymphoproliferative disorders (PTLD), Vitamin E (Vit. E)",
author = "Changguo Chen and Reddy, {Kunam Sudhakar} and Johnston, {T. D.} and Khan, {T. T.} and D. Ranjan",
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T1 - Vitamin E inhibits cyclosporin A and H2O2 promoted Epstein-Barr virus (EBV) transformation of human B cells as assayed by EBV oncogene LMP1 expression

AU - Chen, Changguo

AU - Reddy, Kunam Sudhakar

AU - Johnston, T. D.

AU - Khan, T. T.

AU - Ranjan, D.

PY - 2003/8

Y1 - 2003/8

N2 - Background. We have previously shown that oxidative stress induced by H2O2 or cyclosporin A (CsA) can promote Epstein-Barr virus (EBV) transformation of human B cells as analyzed by colony formation, cell number, and by 3H-thymidine incorporation. In this report, we used EBV oncogene LMP1 as a marker to analyze H2O2 or CsA promotion of EBV transformation of human B cells and to test whether antioxidant vitamin E could inhibit H2O2 or CsA promoted LMP1 expression in the EBV-infected cells. Materials and methods. Human splenocytes were prepared by centrifugation and plating technique to provide a greater than 80% pure preparation of B cells and were used for EBV infection. The EBV infected cells were treated with H2O2 (0.1 mM, 10 min), or with CsA (500 ng/ml) with or with out vitamin E (40 μM). The cells were cultured for up to 4 weeks. Samples were taken every week and were stained with phycoerythrin-conjugated mouse anti-LMP1 monoclonal antibody to assay LMP1 positive population by flow cytometry. Results. In EBV-infected cells, the LMP1-positive cell population reached 14% after 4 weeks of culture. CsA or H2O2 treatment promoted LMP1 positive population to 43% and 41% after 4 weeks of culture. Vitamin E (40 μM) completely inhibited LMP1 expression in EBV-infected cells and in CsA- or H2O 2-treated cells. Conclusion. In agreement with our previous observation, CsA or H2O2 can promote EBV transformation of human B cells. This oxidative stress induced promotion of EBV transformation can be blocked by antioxidant Vitamin E. This finding may have future therapeutic implications for post-transplant lymphoproliferative disorder.

AB - Background. We have previously shown that oxidative stress induced by H2O2 or cyclosporin A (CsA) can promote Epstein-Barr virus (EBV) transformation of human B cells as analyzed by colony formation, cell number, and by 3H-thymidine incorporation. In this report, we used EBV oncogene LMP1 as a marker to analyze H2O2 or CsA promotion of EBV transformation of human B cells and to test whether antioxidant vitamin E could inhibit H2O2 or CsA promoted LMP1 expression in the EBV-infected cells. Materials and methods. Human splenocytes were prepared by centrifugation and plating technique to provide a greater than 80% pure preparation of B cells and were used for EBV infection. The EBV infected cells were treated with H2O2 (0.1 mM, 10 min), or with CsA (500 ng/ml) with or with out vitamin E (40 μM). The cells were cultured for up to 4 weeks. Samples were taken every week and were stained with phycoerythrin-conjugated mouse anti-LMP1 monoclonal antibody to assay LMP1 positive population by flow cytometry. Results. In EBV-infected cells, the LMP1-positive cell population reached 14% after 4 weeks of culture. CsA or H2O2 treatment promoted LMP1 positive population to 43% and 41% after 4 weeks of culture. Vitamin E (40 μM) completely inhibited LMP1 expression in EBV-infected cells and in CsA- or H2O 2-treated cells. Conclusion. In agreement with our previous observation, CsA or H2O2 can promote EBV transformation of human B cells. This oxidative stress induced promotion of EBV transformation can be blocked by antioxidant Vitamin E. This finding may have future therapeutic implications for post-transplant lymphoproliferative disorder.

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KW - Epstein-Barr virus

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KW - Oxidative stress

KW - Post-transplant lymphoproliferative disorders (PTLD)

KW - Vitamin E (Vit. E)

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