Analysis of antiviral drug properties of thymidine kinase of herpes B virus using recombinant herpes simplex virus 1

Abstract

ABSTRACT Zoonotic infection by herpes B virus (BV) causes severe neurological disease, which in the absence of appropriate treatment can have a mortality rate of 80%. Long-term administration of acyclovir or ganciclovir is the recommended prophylactic treatment option for BV infections. However, the emergence of acyclovir- and/or ganciclovir-resistant BV may become a major problem. Here, we examined the ability of BV thymidine kinase (TK) from monkeys (mBVTK) and humans (hBVTK) to acquire resistance to acyclovir and ganciclovir. The sequences of mBVTK and hBVTK differ by two amino acids. We constructed and expressed four HSV-1-based recombinant viruses (HSV-1_mBVTK, HSV-1_hBVTK, HSV-1_VZVTK, and HSV_HSV-1TK) in which the HSV-1TK gene was deleted and the TK gene of monkey BV, human BV, varicella-zoster virus, and HSV-1 was inserted, respectively. We then tested the sensitivity of these recombinant viruses to acyclovir, ganciclovir, penciclovir, and brivudine. HSV-1_HSV-1TK and wild-type HSV-1 were more susceptible to ganciclovir, acyclovir, and penciclovir than HSV-1_VZVTK, HSV-1_hBVTK, and HSV-1_mBVTK. Both HSV-1_hBVTK and HSV-1_mBVTK exhibited similar sensitivity profiles against all tested drugs: both were susceptible to acyclovir, ganciclovir, and penciclovir but resistant to brivudine. In addition, we forced HSV-1_hBVTK to replicate under selective pressure from acyclovir or ganciclovir and then examined the ability of the BV TK gene to develop drug resistance. Investigation of the drug-resistant HSV-1_hBVTK gene revealed the absence of amino acid substitutions in BV TK, suggesting that the development of acyclovir- or ganciclovir-resistant BV through mutations in the TK gene is a rare occurrence in this experimental system. IMPORTANCE Zoonotic infection of humans with herpes B virus (BV) causes severe neurological diseases. Acyclovir (ACV) and ganciclovir (GCV), most frequently used as anti-herpes drugs, are recommended for prophylaxis and therapy in human BV infection. In this study, we examined the property of BV thymidine kinase (TK) against anti-herpes drugs using a recombinant herpes simplex virus type 1 (HSV-1) carrying BV TK gene. We found that HSV-1 carrying BV TK was similarly sensitive to GCV as HSV-1 carrying varicella zoster virus TK. In addition, we demonstrated that BV TK was not mutated in the GCV- and ACV-resistant HSV-1 carrying BV TK, suggesting that ACV- or GCV-resistant BV might be rare during treatment with these antiviral drugs. These data can provide a new insight into the properties of BV TK in terms of the development of drug resistance.