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Indole-3-Carbinol Inhibition of Herpes Simplex Virus Replication

Stoner, Terri Dorene
http://rave.ohiolink.edu/etdc/view?acc_num=kent1228328838

Abstract Details

2008, PHD, Kent State University, College of Arts and Sciences / School of Biomedical Sciences.
Indole-3-carbinol (I3C) is a natural component of a variety of cruciferous plants including broccoli, cabbage, and brussel sprouts. I3C is an antagonist of the G1 phase of the cell cycle (Cover et al., JBC 1998; 273: 3838-47). It is known that cell cycle factors are required for herpes simplex virus (HSV) to replicate (Hossain et al., J. Gen. Virol. 1997; 8: 3341-48). We have established that this relatively noncytotoxic phytocompound inhibits replication of HSV in vitro in a dose and time dependent manner probably by adversely affecting cell cycle factors. To determine the effect of I3C on viral replication, Vero cells (monkey kidney) and MRC-5 cells (human lung fibroblast) were pretreated with noncytotoxic concentrations of 267µM and 375µM I3C, respectively. Cells were infected with HSV-1, HSV-2 or acyclovir resistant HSV-1 at a multiplicity of infection of one and viral yields were assayed. Although, replication of all HSV types tested was inhibited by at least 99.9% by the drug, there were cell line specific differences. Vero cells required pretreatment with I3C for at least 12 hours and MRC-5 cells required pretreatment for 36 hours prior to infection. The observed inhibition was not due to direct viral inactivation or drug induced cytotoxicity. Cell cycle factors were analyzed in MRC-5 cells in the presence or absence of I3C using Western blot and macroarray gene analysis. Western Blot analysis revealed that MRC-5 cells pretreated with I3C for 24 and 36 hours showed significant increases in p53 and p21. HSV-1 infected cells pretreated with I3C demonstrated elevated levels of p53. These results were supported by macroarray gene analysis which demonstrated increased mRNA levels of p53 responsive genes such as p21 and mdm2 after I3C exposure. Western blot analysis of cells pretreated with I3C and infected with HSV showed that Cdk2 protein levels were not significantly affected but host cell cycle factor, cyclin A levels diminished with drug treatment. Cyclin D1 levels were not affected in cells pretreated and HSV infected, but levels of cyclin D3 which is known to interact, stabilize and bind to HSV immediate-early protein (IE), ICP0 (Van Sant et al., J. Virol. 2001; 75:1888-98) increased slightly in comparison to untreated and infected control MRC-5 cells. When the expression of IE ICP-4, the major HSV transactivating protein, was examined its levels were found not to be affected in pretreated and HSV-1 infected MRC-5 cells when compared to controls. Thus the observed inhibition of HSV due to I3C treatment may not involve ICP-4. Therefore, the inhibition exerted by I3C should occur at a point later in the viral replication cycle and after IE proteins are synthesized that are necessary for HSV-1 replication. It appears that I3C may act by disrupting cell cycle factors required by HSV for replication by manipulating p53 and p21. I3C treatment also appears to influence expression of cyclins A and D3 which are known to be involved in the G1/S transition of the host cell cycle and known to be utilized by HSV-1. These results suggest that I3C has significant anti-HSV properties in vitro and the observed inhibition occurs during the G1/S phase of the host cell cycle when HSV-1 DNA synthesis occurs.
John Docherty, PhD (Advisor)
Angelo DeLucia, PhD (Committee Member)
Kenneth Rosenthal, PhD (Committee Member)
Mark Simmons, PhD (Committee Member)
Brent Bruot, PhD (Committee Member)
Biomedical Research
Indole-3-Carbinol; Herpes Simplex Virus; antiviral; broccoli; G1/S; I3C; cell cycle; cyclin dependent kinase; cyclin; MRC-5 cells; HSV; cruciferous; phytocompound

Recommended Citations

Citations

  • Stoner, T. D. (2008). Indole-3-Carbinol Inhibition of Herpes Simplex Virus Replication [Doctoral dissertation, Kent State University]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=kent1228328838

    APA Style (7th edition)

  • Stoner, Terri. Indole-3-Carbinol Inhibition of Herpes Simplex Virus Replication. 2008. Kent State University, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=kent1228328838.

    MLA Style (8th edition)

  • Stoner, Terri. "Indole-3-Carbinol Inhibition of Herpes Simplex Virus Replication." Doctoral dissertation, Kent State University, 2008. http://rave.ohiolink.edu/etdc/view?acc_num=kent1228328838

    Chicago Manual of Style (17th edition)

kent1228328838
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This open access ETD is published by Kent State University and OhioLINK.