T. Suzutani - Academia.edu (original) (raw)
Papers by T. Suzutani
[Hokkaido igaku zasshi] The Hokkaido journal of medical science, 1978
The authors have investigated ten kinds of putrefactive findings on 368 cadavers which were subje... more The authors have investigated ten kinds of putrefactive findings on 368 cadavers which were subjected to medico-legal autopsies in our laboratory and have come to the following conclusions. 1) Concerning the occurrence of the putrefactive findings, seasonal variation was not seen on the cadavers which had remained indoors in the Hokkaido districts. No variation due to age and sex was seen. The occurrence of the findings was less frequent in death from exsanguination, traumatic shock and cardiac tamponade. 2) The putrefactive findings can be utilized as a criterion for estimating the postmortem interval; the interval is estimated to be over 6 hours when the pericardial fluid is turbid, over 12 hours when exudation of putrefactive fluid, gas formation, blood-stained peritoneal fluid or blood-stained pericardial fluid is present, over 18 hours when greenish discoloration of the skin, network of the skin or blood staining of the endocardium or the endothelium is seen, over 2 days when b...
[Hokkaido igaku zasshi] The Hokkaido journal of medical science, 1977
The authors have calculated the amounts of solid matters from the values of water contents determ... more The authors have calculated the amounts of solid matters from the values of water contents determined on 14 sorts of tissues of 41 cadavers and have found that the variation of amounts of solid matters among the same sort of tissue are very big. Accordingly it may be rational to express the values of concentration based on dry weight as to the substances mainly bound to solid matters in tissues and based on fresh weight as to those mainly dissolved in water in tissues.
Journal of Clinical Microbiology, 2000
A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella-zoster virus (VZ... more A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella-zoster virus (VZV) thymidine kinase (TK) genes was developed for monitoring acyclovir-resistant viruses. This method determines the biochemical phenotype of the TK polypeptide, which is synthesized in vitro from viral DNA using a procedure as follows. The TK gene of each sample virus strain is amplified and isolated under the control of a T7 promoter by PCR. The PCR products are transcribed with T7 RNA polymerase and translated in a rabbit reticulocyte lysate. Using this method, enzymatic characteristics and the size of the TK polypeptides encoding HSV and VZV DNA were defined in less than 2 days without virus isolation. The assay should be a powerful tool in monitoring drug-resistant viruses, especially in cases in which virus isolation is difficult.
Acta virologica, 1992
PR-RK, a cell line derived from rabbit kidney cells (RK-13), was insensitive to the cytotoxic eff... more PR-RK, a cell line derived from rabbit kidney cells (RK-13), was insensitive to the cytotoxic effect and interferon (IFN) inducing activity of the copolymer of riboinosinic and ribocytidylic acid (poly rI:rC). However, PR-RK was sensitive to the cytotoxic effect of the copolymer of riboadenylic and ribouridylic acid (poly rA:rU). Comparison of PR-RK cells and RK-13 cells by cytofluorometric analysis revealed that the binding of poly rI:rC was considerably reduced on PR-RK cells. These results suggested that the receptor for poly rI:rC might be different from the receptor for poly rA:rU, and this difference could provide a basis for the identification of the dsRNA receptor on cell surface. Western blot analysis of the components of cell membrane fraction prepared from RK-13 cells was performed by using a monoclonal antibody, which binds to cell membrane of RK-13 cells but not to PR-RK cells, and which blocks IFN induction by poly rI:rC in RK-13 cells. The 60K protein was identified a...
The Tohoku Journal of Experimental Medicine, 1992
9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV) and novel nucleosides, 9-(2-deoxy-2-hydroxymeth... more 9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV) and novel nucleosides, 9-(2-deoxy-2-hydroxymethyl-/3-D-erythro-oxetanocyl)guanine (oxetanocin-G, OXT-G) and (+)-9-[(1R, 2R, 3S)-2, 3-bis(hydroxymethyl)cyclobutyl] guanine (carbocyclic oxetanocin-G, carbocyclic OXT-G) possessed substantial antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). ACV inhibited only viral thymidine kinase positive (TK+) herpes viruses, although the latter two compounds inhibited the replications of the TK deficient (TK-) mutants of HSV-1 and HSV-2 as well as the TK+ parent strains in vitro. The TK-mutants of HSV-1 and HSV-2 (HSV-1 TK-and HSV-2 TK-) were as susceptible to OXT-G as the TK parent strains. However, the TKmutants were less susceptible to carbocyclic OXT-G than the TK+ parent strains. We demonstrated synergistic inhibition of the replications of HSV-1 and HSV-2 by ACV and OXT-G in combination, additive inhibition of HSV-1 and HSV-2 by ACV and carbocyclic OXT-G in combination, synergistic inhibition of HSV-1 by OXT-G and carbocyclic OXT-G in combination, and additive inhibition of HSV-2 by these two compounds. We investigated the metabolism of ACV and OXT-G in HSV-1 TK+-, HSV-1 TK-and mock-infected Vero cells by thin layer chromatography. ACV-triphosphate increased more in HSV-1 TK+-infected Vero cells than in HSV-1 TK-and mock-infected Vero cells. The metabolism of OXT-G had almost the same pattern in HSV-1 TK+-, HSV-1 TK-and mock-infected Vero cells. These results suggest that ACV is phosphorylated by virus-induced TK, and OXT-G is phosphorylated by cellular nucleoside and nucleotide kinases. oxetanocin-G ; carbocyclic oxetanocin-G ; ACV ; HSV-1; HSV-2 Herpes simplex virus (HSV) infections such as genital herpes and neonatal HSV infection appear to be increasing (Sullivan-Bolyai et al. 1983) and continue to cause considerable mortality and morbidity (Whitley et al. 1980; Nahmias et
The Journal of Pediatrics, 2009
A retrospective diagnosis of congenital cytomegalovirus infection was made for three (12%) of 26 ... more A retrospective diagnosis of congenital cytomegalovirus infection was made for three (12%) of 26 students with either bilateral profound or severe sensorineural hearing loss at a School for the Deaf in Japan by detecting viral DNA with real-time PCR from dried umbilical cords that had been preserved at home.
Journal of Medical Virology, 2002
We previously isolated an acyclovir (ACV)-resistant herpes simplex virus type 1 (HSV-1), strain T... more We previously isolated an acyclovir (ACV)-resistant herpes simplex virus type 1 (HSV-1), strain TAR, from a child with Wiskott-Aldrich syndrome. An acyclovir-sensitive HSV-1, strain TAS, had been isolated from the same patient before the isolation of HSV-1 TAR. The TK protein of ACV-sensitive HSV-1 TAS was composed of 376 amino acids, while that of HSV-1 TAR was composed of 407 amino acids with altered amino acid residue between positions 355-407. The elongation of TK was caused by a single nucleotide deletion of cytosine from a homopolymer stretch of 4 cytosine residues between positions 1061-1064. There was no viral TK activity in HSV-1 TAR-infected Vero cells, indicating the importance of the C-terminal portion of TK protein from positions 355-376. Recombinant TK polypeptides with amino acid deletions at the C-terminus were prepared, and TK and ACV-phosphorylation activities were examined. Deletion of 5 and 6 amino acids from the C-terminus of the TK polypeptide of HSV-1 TAS resulted in a reduction of TK activity by approximately 75% and 100%, respectively. These mutant TK polypeptides did not phosphorylate ACV. These results indicate that amino acid residues from positions 371-376 in the C-terminal portion of HSV-1 TK protein are essential for keeping TK and ACV-phosphorylation activities.
Archives of Virology, 1997
Archives of Virology, 2007
A herpes simplex virus type 1 (HSV-1) containing a thymidine (TK) gene with an amber mutation at ... more A herpes simplex virus type 1 (HSV-1) containing a thymidine (TK) gene with an amber mutation at the 8th position counted from the first AUG codon was isolated from a child with acute gingivostomatitis. The virus was predicted to express a mutant viral translated from the 2nd AUG codon at the 46th amino acid position and consisting of 331 amino acids. The virus was as sensitive to acyclovir (ACV), 5-bromovinyl-2'-deoxyuridine (BVdU), 1-beta-D-arabinofuranosyl-(E)-5-(2-bromovinyl)uracil (BVaraU), and 1-beta-D-arabinofuranosylthymine (araT) as a wild-type HSV-1. The mutant TK showed the same level of TK activity as the wild-type TK at reaction temperatures of 34 degrees C, 37 degrees C and 39 degrees C. ACV, BVdU, BVaraU, and araT inhibited the replication of the TK-deficient and drug-resistant HSV-1 and HSV-2 in 293T cells in which the mutant TK was expressed to the same extent as in cells in which intact HSV-1-TK was expressed, whereas BVdU and BVaraU inhibited the replication of these viruses less strongly in cells in which HSV-2-TK was expressed. It can be concluded that the mutant HSV-1 exists in nature as a variant and possesses the necessary phosphorylation activities to form ACV-monophosphate from ACV, to form BVdU-diphosphate through BVdU-monophosphate from BVdU, and to form BVaraU-diphosphate through BVaraU-monophosphate from BVaraU. These results indicate that the mutant HSV-1-TK with a deletion of the first 45 amino acid residues is phenotypically the same as that of wild-type HSV-1-TK in terms of the phosphorylation activity of TK-associated anti-herpes virus drugs.
Antimicrobial Agents and Chemotherapy, 1991
Several new nucleosides with an oxetanosyl-N-glycoside group, named oxetanocins, were evaluated f... more Several new nucleosides with an oxetanosyl-N-glycoside group, named oxetanocins, were evaluated for their antiviral activities against varicella-zoster virus (VZV) in human embryo lung cells. 9-(2-deoxy-2-hydroxy-methyl-beta-D-erythro-oxetanosyl)guanine (OXT-G) and 9-(2-deoxy-2-hydroxymethyl-beta-D-erythro-oxetanosyl)-2- aminoadenine were effective against not only thymidine kinase-positive (TK+) VZV (YS strain) but also thymidine kinase-negative (TK-) VZV (YSR strain), whereas carbocyclic OXT-G was effective against TK+ VZV but not against TK- VZV. [3H]OXT-G was incorporated into TK+ VZV-infected cells and TK- VZV-infected cells more than into mock-infected cells and was converted into the triphosphate form.
Journal of General Virology, 1987
Acta virologica, 1998
To analyze the difference in the degree of divergence between genes from identical herpesvirus sp... more To analyze the difference in the degree of divergence between genes from identical herpesvirus species, we examined the nucleotide sequence of genes from the herpes simplex virus type 1 (HSV-1) strains VR-3 and 17 encoding thymidine kinase (TK), deoxyribonuclease (DNase), protein kinase (PK; UL13) and virion-associated host shutoff (vhs) protein (UL41). The frequency of nucleotide substitutions per 1 kb in TK gene was 2.5 to 4.3 times higher than those in the other three genes. To prove that the polymorphism of HSV-1 TK gene is common characteristic of herpesvirus TK genes, we compared the diversity of TK genes among eight HSV-1, six herpes simplex virus type 2 (HSV-2) and seven varicella-zoster virus (VZV) strains. The average frequency of nucleotide substitutions per 1 kb in the TK gene of HSV-1 strains was 4-fold higher than that in the TK gene of HSV-2 strains. The VZV TK gene was highly conserved and only two nucleotide changes were evident in VZV strains. However, the rate of ...
Antiviral Research, 2000
To clarify whether varicella-zoster virus (VZV) protein kinase (PK; ORF47) takes part in phosphor... more To clarify whether varicella-zoster virus (VZV) protein kinase (PK; ORF47) takes part in phosphorylation of anti-herpesvirus nucleosides, thymidine kinase (TK) deficient, and PK/TK double deficient recombinant VZV strains were isolated and their susceptibility, and that of wild type and PK-deficient strains to various nucleoside analogs was evaluated. The PK-deficient VZV strains showed a sensitivity equal to that of the wild type strain against all compounds tested, including ganciclovir. This indicates that PK is not involved in phosphorylation of the tested nucleosides in VZV-infected cells.
We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simp... more We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simplex virus type 1 (HSV-1) strain VR3. Data from reporter gene assays showed that expression of both type I and type II IFN-inducible genes was dramatically suppressed during the early stage of HSV-1 infection (2 to 3 h postinfection). During these periods, phosphorylation levels of janus kinases (JAKs) and STATs did not increase after treatment of HSV-1-infected FL cells with IFN-␣ or IFN-␥, although cellular protein levels of the JAKs and the STATs were not significantly changed. In contrast, the inhibitory effect of HSV-1 on phosphorylation of STAT1 was not observed in U937 cells, which show resistance to steady-state accumulation of RNA for HSV-1 immediate-early genes. The phosphorylation of STAT1 in FL cells was not inhibited by infection with a UV-inactivated virus. These results indicate that viral gene expression or viral protein production is necessary for the inhibition of phosphorylation by HSV-1.
Journal of General Virology, 1997
Trehalose 6,6h-dimycolate (TDM), an immunomodulator, potentiates non-specific resistance in mice ... more Trehalose 6,6h-dimycolate (TDM), an immunomodulator, potentiates non-specific resistance in mice to influenza virus infection. When mice were injected intravenously with TDM, the striking proliferation of a minority of T-lymphocytes bearing gamma/delta T-cell receptors (γδ T-cells) that accumulated in granulomatous lungs was thought to be associated with the maintenance of acquired resistance to lethal influenza virus infection. To clarify the cellular basis of the defence against influenza virus, mice were depleted of γδ T-cells, alpha/beta (αβ) T-cells, or natural killer (NK) cells by in vivo administration of corresponding antibodies prior to influenza virus infection. The depletion of γδ T-cells significantly abrogated the augmented resistance of TDMtreated mice to infection, as did depletion of either αβ T-cells or NK cells. To gain insight into the
ChemInform, 1996
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Journal of biological response modifiers, 1988
Mice inoculated intravenously with trehalose-6,6'-dimycolate (TDM), a glycolipid component of... more Mice inoculated intravenously with trehalose-6,6'-dimycolate (TDM), a glycolipid component of the cell wall of Mycobacterium, in an oil-in-water emulsion (TDM emulsion) acquired a high resistance to intranasal infection by influenza virus. Athymic nude mice inoculated with TDM emulsion could not acquire such an augmented resistance to influenza virus infection. The augmented antiviral resistance of TDM emulsion-treated mice was diminished by prior intravenous inoculation of silica particles, which selectively impair macrophage functions. In vitro experiments showed that macrophage cultures treated with TDM emulsion released an activator(s) of T lymphocytes. Histological studies of the lung of TDM emulsion-inoculated mice revealed that a typical granuloma and severe perivascular lymphocyte infiltration appeared, though no such histological change was observed in the lung of control emulsion-inoculated mice. The lungs from TDM emulsion-treated athymic nude mice and the lungs from ...
Transplant infectious disease : an official journal of the Transplantation Society, 2011
Cytomegalovirus (CMV) reinfection of seropositive individuals has been associated with adverse ou... more Cytomegalovirus (CMV) reinfection of seropositive individuals has been associated with adverse outcomes in organ transplantation and is a frequent cause of congenital infection. Previously we demonstrated that mismatching of CMV glycoprotein H (gH) serotypes was associated with CMV disease after renal transplantation. Because the antigen domain 2 (AD2) epitope of glycoprotein B (gB) is conserved among CMV isolates and is one of the known targets of neutralizing antibodies, in this study we investigated whether antibodies against the epitope contribute to protection from CMV reinfection in renal transplantation, irrespective of gH serological matching. For this purpose, the gB and gH serology and clinical outcomes were analyzed retrospectively for 77 transplant recipients in the donor positive/recipient positive setting, who were managed by preemptive strategy. We found that there was a good negative correlation between the numbers of antigenemia-positive cells and the levels of anti...
Acta virologica, 2001
Earlier, we have detected antiviral activity in an extract from Ribes nigrum L. fruits ("Kur... more Earlier, we have detected antiviral activity in an extract from Ribes nigrum L. fruits ("Kurokarin", name of the one species of black currant in Japanese) against influenza A and B viruses, and herpes simplex virus 1 (Knox et al., Food Processing 33, 21-23, 1998). In the present study, the antiviral activity of constituents of a Kurokarin extract and the mechanism of its antiviral action were examined. Kurokarin extracts were separated to fractions A to D by column chromatography. The major constituents of the fraction D were estimated as anthocyanins. The fraction D was further fractionated by thin-layer chromatography (TLC) to fractions A' to G'. The fraction E' consisted of 3-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl-cyanidin and 3-O-beta-D-glucopyranosyl-cyanidin, and the fraction F' consisted of 3-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl-delphinidin and 3-O-beta-D-glucopyranosyl-delphinidin, identified by high performance liquid chromatogr...
Virology, Jan 20, 2001
We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simp... more We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simplex virus type 1 (HSV-1) strain VR3. Data from reporter gene assays showed that expression of both type I and type II IFN-inducible genes was dramatically suppressed during the early stage of HSV-1 infection (2 to 3 h postinfection). During these periods, phosphorylation levels of janus kinases (JAKs) and STATs did not increase after treatment of HSV-1-infected FL cells with IFN-alpha or IFN-gamma, although cellular protein levels of the JAKs and the STATs were not significantly changed. In contrast, the inhibitory effect of HSV-1 on phosphorylation of STAT1 was not observed in U937 cells, which show resistance to steady-state accumulation of RNA for HSV-1 immediate-early genes. The phosphorylation of STAT1 in FL cells was not inhibited by infection with a UV-inactivated virus. These results indicate that viral gene expression or viral protein production is necessary for the inhibition o...
[Hokkaido igaku zasshi] The Hokkaido journal of medical science, 1978
The authors have investigated ten kinds of putrefactive findings on 368 cadavers which were subje... more The authors have investigated ten kinds of putrefactive findings on 368 cadavers which were subjected to medico-legal autopsies in our laboratory and have come to the following conclusions. 1) Concerning the occurrence of the putrefactive findings, seasonal variation was not seen on the cadavers which had remained indoors in the Hokkaido districts. No variation due to age and sex was seen. The occurrence of the findings was less frequent in death from exsanguination, traumatic shock and cardiac tamponade. 2) The putrefactive findings can be utilized as a criterion for estimating the postmortem interval; the interval is estimated to be over 6 hours when the pericardial fluid is turbid, over 12 hours when exudation of putrefactive fluid, gas formation, blood-stained peritoneal fluid or blood-stained pericardial fluid is present, over 18 hours when greenish discoloration of the skin, network of the skin or blood staining of the endocardium or the endothelium is seen, over 2 days when b...
[Hokkaido igaku zasshi] The Hokkaido journal of medical science, 1977
The authors have calculated the amounts of solid matters from the values of water contents determ... more The authors have calculated the amounts of solid matters from the values of water contents determined on 14 sorts of tissues of 41 cadavers and have found that the variation of amounts of solid matters among the same sort of tissue are very big. Accordingly it may be rational to express the values of concentration based on dry weight as to the substances mainly bound to solid matters in tissues and based on fresh weight as to those mainly dissolved in water in tissues.
Journal of Clinical Microbiology, 2000
A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella-zoster virus (VZ... more A rapid phenotypic screening method for herpes simplex virus (HSV) and varicella-zoster virus (VZV) thymidine kinase (TK) genes was developed for monitoring acyclovir-resistant viruses. This method determines the biochemical phenotype of the TK polypeptide, which is synthesized in vitro from viral DNA using a procedure as follows. The TK gene of each sample virus strain is amplified and isolated under the control of a T7 promoter by PCR. The PCR products are transcribed with T7 RNA polymerase and translated in a rabbit reticulocyte lysate. Using this method, enzymatic characteristics and the size of the TK polypeptides encoding HSV and VZV DNA were defined in less than 2 days without virus isolation. The assay should be a powerful tool in monitoring drug-resistant viruses, especially in cases in which virus isolation is difficult.
Acta virologica, 1992
PR-RK, a cell line derived from rabbit kidney cells (RK-13), was insensitive to the cytotoxic eff... more PR-RK, a cell line derived from rabbit kidney cells (RK-13), was insensitive to the cytotoxic effect and interferon (IFN) inducing activity of the copolymer of riboinosinic and ribocytidylic acid (poly rI:rC). However, PR-RK was sensitive to the cytotoxic effect of the copolymer of riboadenylic and ribouridylic acid (poly rA:rU). Comparison of PR-RK cells and RK-13 cells by cytofluorometric analysis revealed that the binding of poly rI:rC was considerably reduced on PR-RK cells. These results suggested that the receptor for poly rI:rC might be different from the receptor for poly rA:rU, and this difference could provide a basis for the identification of the dsRNA receptor on cell surface. Western blot analysis of the components of cell membrane fraction prepared from RK-13 cells was performed by using a monoclonal antibody, which binds to cell membrane of RK-13 cells but not to PR-RK cells, and which blocks IFN induction by poly rI:rC in RK-13 cells. The 60K protein was identified a...
The Tohoku Journal of Experimental Medicine, 1992
9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV) and novel nucleosides, 9-(2-deoxy-2-hydroxymeth... more 9-(2-hydroxyethoxymethyl)guanine (acyclovir, ACV) and novel nucleosides, 9-(2-deoxy-2-hydroxymethyl-/3-D-erythro-oxetanocyl)guanine (oxetanocin-G, OXT-G) and (+)-9-[(1R, 2R, 3S)-2, 3-bis(hydroxymethyl)cyclobutyl] guanine (carbocyclic oxetanocin-G, carbocyclic OXT-G) possessed substantial antiviral activities against herpes simplex virus type 1 (HSV-1) and type 2 (HSV-2). ACV inhibited only viral thymidine kinase positive (TK+) herpes viruses, although the latter two compounds inhibited the replications of the TK deficient (TK-) mutants of HSV-1 and HSV-2 as well as the TK+ parent strains in vitro. The TK-mutants of HSV-1 and HSV-2 (HSV-1 TK-and HSV-2 TK-) were as susceptible to OXT-G as the TK parent strains. However, the TKmutants were less susceptible to carbocyclic OXT-G than the TK+ parent strains. We demonstrated synergistic inhibition of the replications of HSV-1 and HSV-2 by ACV and OXT-G in combination, additive inhibition of HSV-1 and HSV-2 by ACV and carbocyclic OXT-G in combination, synergistic inhibition of HSV-1 by OXT-G and carbocyclic OXT-G in combination, and additive inhibition of HSV-2 by these two compounds. We investigated the metabolism of ACV and OXT-G in HSV-1 TK+-, HSV-1 TK-and mock-infected Vero cells by thin layer chromatography. ACV-triphosphate increased more in HSV-1 TK+-infected Vero cells than in HSV-1 TK-and mock-infected Vero cells. The metabolism of OXT-G had almost the same pattern in HSV-1 TK+-, HSV-1 TK-and mock-infected Vero cells. These results suggest that ACV is phosphorylated by virus-induced TK, and OXT-G is phosphorylated by cellular nucleoside and nucleotide kinases. oxetanocin-G ; carbocyclic oxetanocin-G ; ACV ; HSV-1; HSV-2 Herpes simplex virus (HSV) infections such as genital herpes and neonatal HSV infection appear to be increasing (Sullivan-Bolyai et al. 1983) and continue to cause considerable mortality and morbidity (Whitley et al. 1980; Nahmias et
The Journal of Pediatrics, 2009
A retrospective diagnosis of congenital cytomegalovirus infection was made for three (12%) of 26 ... more A retrospective diagnosis of congenital cytomegalovirus infection was made for three (12%) of 26 students with either bilateral profound or severe sensorineural hearing loss at a School for the Deaf in Japan by detecting viral DNA with real-time PCR from dried umbilical cords that had been preserved at home.
Journal of Medical Virology, 2002
We previously isolated an acyclovir (ACV)-resistant herpes simplex virus type 1 (HSV-1), strain T... more We previously isolated an acyclovir (ACV)-resistant herpes simplex virus type 1 (HSV-1), strain TAR, from a child with Wiskott-Aldrich syndrome. An acyclovir-sensitive HSV-1, strain TAS, had been isolated from the same patient before the isolation of HSV-1 TAR. The TK protein of ACV-sensitive HSV-1 TAS was composed of 376 amino acids, while that of HSV-1 TAR was composed of 407 amino acids with altered amino acid residue between positions 355-407. The elongation of TK was caused by a single nucleotide deletion of cytosine from a homopolymer stretch of 4 cytosine residues between positions 1061-1064. There was no viral TK activity in HSV-1 TAR-infected Vero cells, indicating the importance of the C-terminal portion of TK protein from positions 355-376. Recombinant TK polypeptides with amino acid deletions at the C-terminus were prepared, and TK and ACV-phosphorylation activities were examined. Deletion of 5 and 6 amino acids from the C-terminus of the TK polypeptide of HSV-1 TAS resulted in a reduction of TK activity by approximately 75% and 100%, respectively. These mutant TK polypeptides did not phosphorylate ACV. These results indicate that amino acid residues from positions 371-376 in the C-terminal portion of HSV-1 TK protein are essential for keeping TK and ACV-phosphorylation activities.
Archives of Virology, 1997
Archives of Virology, 2007
A herpes simplex virus type 1 (HSV-1) containing a thymidine (TK) gene with an amber mutation at ... more A herpes simplex virus type 1 (HSV-1) containing a thymidine (TK) gene with an amber mutation at the 8th position counted from the first AUG codon was isolated from a child with acute gingivostomatitis. The virus was predicted to express a mutant viral translated from the 2nd AUG codon at the 46th amino acid position and consisting of 331 amino acids. The virus was as sensitive to acyclovir (ACV), 5-bromovinyl-2'-deoxyuridine (BVdU), 1-beta-D-arabinofuranosyl-(E)-5-(2-bromovinyl)uracil (BVaraU), and 1-beta-D-arabinofuranosylthymine (araT) as a wild-type HSV-1. The mutant TK showed the same level of TK activity as the wild-type TK at reaction temperatures of 34 degrees C, 37 degrees C and 39 degrees C. ACV, BVdU, BVaraU, and araT inhibited the replication of the TK-deficient and drug-resistant HSV-1 and HSV-2 in 293T cells in which the mutant TK was expressed to the same extent as in cells in which intact HSV-1-TK was expressed, whereas BVdU and BVaraU inhibited the replication of these viruses less strongly in cells in which HSV-2-TK was expressed. It can be concluded that the mutant HSV-1 exists in nature as a variant and possesses the necessary phosphorylation activities to form ACV-monophosphate from ACV, to form BVdU-diphosphate through BVdU-monophosphate from BVdU, and to form BVaraU-diphosphate through BVaraU-monophosphate from BVaraU. These results indicate that the mutant HSV-1-TK with a deletion of the first 45 amino acid residues is phenotypically the same as that of wild-type HSV-1-TK in terms of the phosphorylation activity of TK-associated anti-herpes virus drugs.
Antimicrobial Agents and Chemotherapy, 1991
Several new nucleosides with an oxetanosyl-N-glycoside group, named oxetanocins, were evaluated f... more Several new nucleosides with an oxetanosyl-N-glycoside group, named oxetanocins, were evaluated for their antiviral activities against varicella-zoster virus (VZV) in human embryo lung cells. 9-(2-deoxy-2-hydroxy-methyl-beta-D-erythro-oxetanosyl)guanine (OXT-G) and 9-(2-deoxy-2-hydroxymethyl-beta-D-erythro-oxetanosyl)-2- aminoadenine were effective against not only thymidine kinase-positive (TK+) VZV (YS strain) but also thymidine kinase-negative (TK-) VZV (YSR strain), whereas carbocyclic OXT-G was effective against TK+ VZV but not against TK- VZV. [3H]OXT-G was incorporated into TK+ VZV-infected cells and TK- VZV-infected cells more than into mock-infected cells and was converted into the triphosphate form.
Journal of General Virology, 1987
Acta virologica, 1998
To analyze the difference in the degree of divergence between genes from identical herpesvirus sp... more To analyze the difference in the degree of divergence between genes from identical herpesvirus species, we examined the nucleotide sequence of genes from the herpes simplex virus type 1 (HSV-1) strains VR-3 and 17 encoding thymidine kinase (TK), deoxyribonuclease (DNase), protein kinase (PK; UL13) and virion-associated host shutoff (vhs) protein (UL41). The frequency of nucleotide substitutions per 1 kb in TK gene was 2.5 to 4.3 times higher than those in the other three genes. To prove that the polymorphism of HSV-1 TK gene is common characteristic of herpesvirus TK genes, we compared the diversity of TK genes among eight HSV-1, six herpes simplex virus type 2 (HSV-2) and seven varicella-zoster virus (VZV) strains. The average frequency of nucleotide substitutions per 1 kb in the TK gene of HSV-1 strains was 4-fold higher than that in the TK gene of HSV-2 strains. The VZV TK gene was highly conserved and only two nucleotide changes were evident in VZV strains. However, the rate of ...
Antiviral Research, 2000
To clarify whether varicella-zoster virus (VZV) protein kinase (PK; ORF47) takes part in phosphor... more To clarify whether varicella-zoster virus (VZV) protein kinase (PK; ORF47) takes part in phosphorylation of anti-herpesvirus nucleosides, thymidine kinase (TK) deficient, and PK/TK double deficient recombinant VZV strains were isolated and their susceptibility, and that of wild type and PK-deficient strains to various nucleoside analogs was evaluated. The PK-deficient VZV strains showed a sensitivity equal to that of the wild type strain against all compounds tested, including ganciclovir. This indicates that PK is not involved in phosphorylation of the tested nucleosides in VZV-infected cells.
We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simp... more We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simplex virus type 1 (HSV-1) strain VR3. Data from reporter gene assays showed that expression of both type I and type II IFN-inducible genes was dramatically suppressed during the early stage of HSV-1 infection (2 to 3 h postinfection). During these periods, phosphorylation levels of janus kinases (JAKs) and STATs did not increase after treatment of HSV-1-infected FL cells with IFN-␣ or IFN-␥, although cellular protein levels of the JAKs and the STATs were not significantly changed. In contrast, the inhibitory effect of HSV-1 on phosphorylation of STAT1 was not observed in U937 cells, which show resistance to steady-state accumulation of RNA for HSV-1 immediate-early genes. The phosphorylation of STAT1 in FL cells was not inhibited by infection with a UV-inactivated virus. These results indicate that viral gene expression or viral protein production is necessary for the inhibition of phosphorylation by HSV-1.
Journal of General Virology, 1997
Trehalose 6,6h-dimycolate (TDM), an immunomodulator, potentiates non-specific resistance in mice ... more Trehalose 6,6h-dimycolate (TDM), an immunomodulator, potentiates non-specific resistance in mice to influenza virus infection. When mice were injected intravenously with TDM, the striking proliferation of a minority of T-lymphocytes bearing gamma/delta T-cell receptors (γδ T-cells) that accumulated in granulomatous lungs was thought to be associated with the maintenance of acquired resistance to lethal influenza virus infection. To clarify the cellular basis of the defence against influenza virus, mice were depleted of γδ T-cells, alpha/beta (αβ) T-cells, or natural killer (NK) cells by in vivo administration of corresponding antibodies prior to influenza virus infection. The depletion of γδ T-cells significantly abrogated the augmented resistance of TDMtreated mice to infection, as did depletion of either αβ T-cells or NK cells. To gain insight into the
ChemInform, 1996
ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance t... more ABSTRACT ChemInform is a weekly Abstracting Service, delivering concise information at a glance that was extracted from about 100 leading journals. To access a ChemInform Abstract of an article which was published elsewhere, please select a “Full Text” option. The original article is trackable via the “References” option.
Journal of biological response modifiers, 1988
Mice inoculated intravenously with trehalose-6,6'-dimycolate (TDM), a glycolipid component of... more Mice inoculated intravenously with trehalose-6,6'-dimycolate (TDM), a glycolipid component of the cell wall of Mycobacterium, in an oil-in-water emulsion (TDM emulsion) acquired a high resistance to intranasal infection by influenza virus. Athymic nude mice inoculated with TDM emulsion could not acquire such an augmented resistance to influenza virus infection. The augmented antiviral resistance of TDM emulsion-treated mice was diminished by prior intravenous inoculation of silica particles, which selectively impair macrophage functions. In vitro experiments showed that macrophage cultures treated with TDM emulsion released an activator(s) of T lymphocytes. Histological studies of the lung of TDM emulsion-inoculated mice revealed that a typical granuloma and severe perivascular lymphocyte infiltration appeared, though no such histological change was observed in the lung of control emulsion-inoculated mice. The lungs from TDM emulsion-treated athymic nude mice and the lungs from ...
Transplant infectious disease : an official journal of the Transplantation Society, 2011
Cytomegalovirus (CMV) reinfection of seropositive individuals has been associated with adverse ou... more Cytomegalovirus (CMV) reinfection of seropositive individuals has been associated with adverse outcomes in organ transplantation and is a frequent cause of congenital infection. Previously we demonstrated that mismatching of CMV glycoprotein H (gH) serotypes was associated with CMV disease after renal transplantation. Because the antigen domain 2 (AD2) epitope of glycoprotein B (gB) is conserved among CMV isolates and is one of the known targets of neutralizing antibodies, in this study we investigated whether antibodies against the epitope contribute to protection from CMV reinfection in renal transplantation, irrespective of gH serological matching. For this purpose, the gB and gH serology and clinical outcomes were analyzed retrospectively for 77 transplant recipients in the donor positive/recipient positive setting, who were managed by preemptive strategy. We found that there was a good negative correlation between the numbers of antigenemia-positive cells and the levels of anti...
Acta virologica, 2001
Earlier, we have detected antiviral activity in an extract from Ribes nigrum L. fruits ("Kur... more Earlier, we have detected antiviral activity in an extract from Ribes nigrum L. fruits ("Kurokarin", name of the one species of black currant in Japanese) against influenza A and B viruses, and herpes simplex virus 1 (Knox et al., Food Processing 33, 21-23, 1998). In the present study, the antiviral activity of constituents of a Kurokarin extract and the mechanism of its antiviral action were examined. Kurokarin extracts were separated to fractions A to D by column chromatography. The major constituents of the fraction D were estimated as anthocyanins. The fraction D was further fractionated by thin-layer chromatography (TLC) to fractions A' to G'. The fraction E' consisted of 3-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl-cyanidin and 3-O-beta-D-glucopyranosyl-cyanidin, and the fraction F' consisted of 3-O-alpha-L-rhamnopyranosyl-beta-D-glucopyranosyl-delphinidin and 3-O-beta-D-glucopyranosyl-delphinidin, identified by high performance liquid chromatogr...
Virology, Jan 20, 2001
We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simp... more We examined the influence on the interferon (IFN) signaling pathway of infection with herpes simplex virus type 1 (HSV-1) strain VR3. Data from reporter gene assays showed that expression of both type I and type II IFN-inducible genes was dramatically suppressed during the early stage of HSV-1 infection (2 to 3 h postinfection). During these periods, phosphorylation levels of janus kinases (JAKs) and STATs did not increase after treatment of HSV-1-infected FL cells with IFN-alpha or IFN-gamma, although cellular protein levels of the JAKs and the STATs were not significantly changed. In contrast, the inhibitory effect of HSV-1 on phosphorylation of STAT1 was not observed in U937 cells, which show resistance to steady-state accumulation of RNA for HSV-1 immediate-early genes. The phosphorylation of STAT1 in FL cells was not inhibited by infection with a UV-inactivated virus. These results indicate that viral gene expression or viral protein production is necessary for the inhibition o...