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Antiviral mechanism(s) of the experimental immunosuppressive agent leflunomide against human cytomegalovirus and polyomavirus

Meister, Gabriel T
http://rave.ohiolink.edu/etdc/view?acc_num=osu1111428519

Abstract Details

2005, Doctor of Philosophy, Ohio State University, Pathology.
Leflunomide is an experimental immunosuppressive agent that has shown efficacy as an antiviral agent against human cytomegalovirus (HCMV) and polyomavirus strain BK (BKV). An antiviral regimen has been approved for immunosuppressed patients suffering complications from HCMV infection, whereas a good treatment option for patients infected with BKV does not exist. Unfortunately, the antiviral treatment options for patients infected with HCMV have helped promote the propagation of multi-drug resistant HCMV strains. This body of work illustrates the possible antiviral mechanisms associated with Leflunomide using an in vitro model system. We have tested the hypothesis that the antiviral activity of A77 1726, the active metabolite of Leflunomide, is a result of its inhibition of phosphorylation of one or more viral structural proteins. Western blot, Southern (Dot Blot) blot, and CMV gene array analysis demonstrated that Leflunomide does not inhibit HCMV DNA synthesis, the translation of essential viral proteins, or the transcription of viral mRNA. 32P-orthophosphate labeling experiments confirm a reduction in the phosphorylation of more than one of the HCMV tegument proteins. In addition, immunohistochemical staining showed discrete changes in localization of these tegument proteins in Leflunomide-treated cells. Co-immunoprecipitation experiments confirm that Leflunomide disrupts the interaction of viral tegument proteins suggesting that Leflunomide may inhibit complete infectious virion assembly by altering the phosphorylation states of one or more viral structural proteins. The second aspect of this work was to determine if Leflunomide would inhibit the replication of BKV, a polyoma virus unrelated to HCMV. We tested the hypothesis that A77 1726 would inhibit the production of infectious BKV particles without inhibiting DNA synthesis or large T antigen translation. Plaque assay data demonstrated a log decrease in viral titers when infected cells were treated with A77 1726. Western blot and Southern blot data confirmed the inhibition was not due to a block in protein translation of the large T antigen or a viral DNA synthesis. Immunohistochemistry confirmed there was no reduction of the large T antigen protein when infected cells were treated with A77 1726. When the phosphorylation of the large T antigen was assessed, no reductions in phosphorylation could be detected.
W. Waldman (Advisor)
141 p.
HCMV; cytomegalovirus; polyomavirus; antivirals; viral replication

Recommended Citations

Citations

  • Meister, G. T. (2005). Antiviral mechanism(s) of the experimental immunosuppressive agent leflunomide against human cytomegalovirus and polyomavirus [Doctoral dissertation, Ohio State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=osu1111428519

    APA Style (7th edition)

  • Meister, Gabriel. Antiviral mechanism(s) of the experimental immunosuppressive agent leflunomide against human cytomegalovirus and polyomavirus. 2005. Ohio State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=osu1111428519.

    MLA Style (8th edition)

  • Meister, Gabriel. "Antiviral mechanism(s) of the experimental immunosuppressive agent leflunomide against human cytomegalovirus and polyomavirus." Doctoral dissertation, Ohio State University, 2005. http://rave.ohiolink.edu/etdc/view?acc_num=osu1111428519

    Chicago Manual of Style (17th edition)

osu1111428519
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This open access ETD is published by The Ohio State University and OhioLINK.