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Herpes Simplex Virus Type 1의 증식기작과 항바이러스물질의 in Vitro 약효평가
이종교,김해수 대한바이러스학회 1992 Journal of Bacteriology and Virology Vol.22 No.2
To delineate the stage of the virus replicative cycle at which antivirals may interact, were performed the measurement of one step growth and viral DNA synthesis curves of HSV-1 strain F and time of addition experiments with 4 antivirals, DS 5000, ACV, Ara-T and FIAC. During one step growth curves maturated viruses appeared in the virus-infected Vero cells after 5h post infection(PI) and reached the highest titer 15h PI. The released viruses started appearing from llh PI and increased until 22h PI. Viral DNA synthesis which was surveyed by radioactivity of incorporated nucleic acid precursors-(H)-thymidine-significantly increased from 7h PI and reached the maximum 15h PI. Time of addition experiment followed by virus yield assay was performed. Virus-in- fected cells were treated at a concentration that was approximately 50-100-fold higher than their EC values. DS 5000, which interferes with virus binding or/and fusion, was no longer able to inhibit virus replication, if added 1h(or later) PI, I.e. after adsorption. ACV, Ara T and FIAC which inhibit viral DNA synthesis were no longer able to inhibit virus replication, if added 1 1h(or later) PI. Time of addition experiment using CPE inhibition assay method to read the results was also performed. The relationship of the increasement of EC of the tested compounds and delayed drug-addition time was very similar to that of the previous experiment. Those results were correlated with one step growth and viral DNA synthesis curves of the virus. It tells us that by controlling the drug addition time(ex, 0, 1, 5, 8, 11 and 15h PI) the stage of virus replicative cycle at which antivirals may interact may be rapidly suggested.
마우스 대뇌감염모델을 이용한 Acyclovir의 항Herpes Simplex Virus Type 1 약효평가
이종교,김해수,Lee, Chong-Kyo,Kim, Hae-Soo 대한미생물학회 1998 Journal of Bacteriology and Virology Vol.39 No.2
To establish in vivo antiviral evaluation system by using murine herpesvirus intracerebral infection model, 5-6 female BALB/c mice per group aged 5 weeks were inoculated i.c. into cerebrum with different inocular HSV-1 F. Signs of clinical disease noted everyday for one month. Observed were body weight decrease, neurological signs and death caused by encephalitis. Mice discontinued body weight decrease were recovered from the disease, and keratitis was often observed during recovery. The groups inoculated with higher than 1,000 PFU showed 100% mortaltiy and $LD_{50}$ was <100 PFU/mouse. To study the effect of virus inoculum sizes on antiviral effect of acyclovir (ACV), mice inoculated with different inocula were administered i.p. with different doses of ACV immediately after infection, and twice a day for 5 days. The higher inculum size, the less protective. $ED_{50}$ of ACV was >25, >25, 18.4 and 8.0 mg/kg b.i.d. in the group infected with 1,000,000, 100,000, 10,000 and 1,000 PFU/mouse, respectively. $LD_{50}$ of ACV was 62.5 mg/kg b.i.d. Therapeutic index of ACV was <2.5, <2.5, 3.0 and 7.0 in the groups with inocula 1,000,000, 100,000, 10,000 and 1,000 PFU/mouse, respectively. Inoculum size 1,000 PFU/mouse showing 100% mortaltiy and 5-6 days mean time to death, 5 days drug administration and 14 days observation will be future experimental conditions.
핵산유도체와 전구물질들의 항 Herpes바이러스 In Vitro 약효평가
이종교,나지숙,김해수 대한바이러스학회 1992 Journal of Bacteriology and Virology Vol.22 No.1
To evaluate in vitro anti herpetic efficacy of nucleoside analogues and precursor (base) analogues, Vero cells were infected with three different strains of HSV-1 and 2 of HSV-1 and 2 of HSV-2 and treated with the compounds. The antiviral activity was evaluated by virus-induced cytopathic effect(CPE) inhibition assay. Anti-CMV activity was measured by CPE inhibition assay and anti-VZV by plaque reduction assay. HEL 299 cells were used for the replication of CMV and VZV. VZV showed generally the highest sensitivity against tested compounds followed by HSV and CMV. There was no significant difference between anti-HSV-1 and anti HSV 2 activity of tested compounds except BVdUrd and TFT. But differences among strains of the same virus were rather observed. All compounds licensed for the clinical treatment, ACV, Ara A, IdUrd, TFT, GCV and PFA, showed in vitro efficacy against their target viruses. Cytostatic and cytocidal effects of the compounds were also measured by MTT assay in Vero cell cultures. The former were more significant than the latter.
마우스 대뇌감염모델을 이용한 Acyclovir의 항 Herpes Simplex Virus Type 1 약효평가
이종교,김해수 대한바이러스학회 1998 Journal of Bacteriology and Virology Vol.28 No.1
To establish in vivo antiviral evaluation system by using murine herpesvirus intracerebral infection model, 5-6 female BALB/c mice per group aged 5 weeks were inoculated I.c. into cerebrum with different inocular HSV-1 F. Signs of clinical disease noted everyday for one month. Observed were body weight decrease, neurological signs and death caused by encephalitis. Mice discontinued body weight decrease were recovered from the disease, and keratitis was often observed during recovery. The groups inoculated with higher than 1,000 PFU showed 100% mortaltiy and LDp was <100 PFU/mouse. To study the effect of virus inoculum sizes on antiviral effect of acyclovir (ACV), mice inoculated with different inocula were administered I.p. with different doses of ACV immediately after infection, and twice a day for 5 days. The higher inculum size, the less protective. Edp of ACV was >25, >25, 18.4 and 8.0 mg/kg b.I.d. in the group infected with 1,000,000, 100,000, 10,000 and 1,000 PFU/mouse, respectively. LD of ACV was 62.5 mg/kg b.I.d. Therapeutic index of ACV was <2.5, <2.5, 3.0 and 7.0 in the groups with inocula 1,000,000, 100,000, 10,000 and 1,000 PFU/mouse, respectively. Inoculum size 1,000 PFU/mouse showing 100% mortaltiy and 5-6 days mean time to death, 5 days drug administration and 14 days observation will be future exeperimental conditions.
핵산유도체들의 항 Human Immunodeficiency Virus in vitro 약효평가와 작용기전연구
이종교,김동기,김지현,김해수,피미경,박종백,김백 대한바이러스학회 1997 Journal of Bacteriology and Virology Vol.27 No.1
To evaluate in vitro anti-HIV efficacies of nucleoside derivatives, MT-4 cell line was infected with HIV-1 and HIV-2 respectively and treated with various compounds and the formerly approved drugs such as AZT, d4T, ddC and ddI. CPE method was used to evaluate their antiviral activity. Most dideoxynucleosides, AZT, d4T, ddC and ddI, showed anti-HIV activities against both viruses but no other compounds including anti-herpesvirus drugs did any. Further experiments were carried out to study their inhibitory mechanism of viral adsorption. The results showed no inhibition of syncytium formation due to an interaction between the gp120 expressed in HIV-infected cell surface and CD4 receptor on the uninfected cell surface in the presence of AZT. AZT showed no activity up to 100 pg/ml. Inhibition of reverse transcriptase (RT) in the presence of AZT-triphosphate was tested by using RT expressed in E. coli and purified and its IC was 4.5 nM.