• Journal of Microbiology and Biotechnology
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• 제23권8호
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• pp.1154-1158
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• 2013

Ganciclovir resistance of human cytomegalovirus is associated with mutations in the viral UL97 gene and poses severe problems for immunocompromised patients. In this study, PCR-based restriction fragment length polymorphism and sequencing analyses detected the UL97 D605E mutation in all five clinical isolates from patients with ganciclovir-resistant human cytomegalovirus infection during prolonged ganciclovir therapy, whereas the M460V mutation was only present in 1 of 5 isolates. On the other hand, the detection rates of the D605E mutation in the stored available DNA samples from the donor and allogeneic stem cell transplantation recipients were 66.7% and 93.7%, respectively, suggesting that the presence of D605E mutation was not associated with the ganciclovir exposure. Although the D605E mutation may not be related to ganciclovir resistance, we suggest that this mutation could be an important molecular marker of human cytomegalovirus evolution in East Asian countries. Moreover, the restriction fragment length polymorphism method using the restriction enzyme HaeIII, which is generally used to detect the UL97 A591V mutation, could also detect the D605E mutation and may therefore be a useful tool for future research on the investigation of UL97 gene mutations.

• 미생물학회지
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• 제29권1호
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• pp.25-33
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• 1991

Antiviral activities of papaverine and nucleoside analogs, 9-[(1,3-dihydroxy-2-propoxy)methyl]guanine (DHPG) and acyclovir, against human cytomegalovirus (HCMV) infection were compared in vitro. Papaverine and DHPG were effective in reducing infectious HCMV yields with $ED_{50}{\s}$ (effective dose 50: the concentraion at which 50% of virus yields was obtained) of approximately 1.02 and $0.45{\mu}{\M}$, respectively; while acyclovir was less effective with an $ED_{50}$ of about $10.4{\mu}{\M}$The relative cytotoxicity of these drugs was evaluated under the same conditions used to measure infectious HCMV yields. Papaverine and DHPG demonstrated little cellular toxicity as measured by their effect on the viability of confluent cells at concentrations in the range of those demonstrating potent inhibition of HCMV replication. Similarly, protein synthesis was largely unaffected by these drugs in stationary mock-infected cells as measured by the incorporation of isotopically labelled amino acids. In contrast, cellular DNA synthesis was invariably reduced in the presence of either drug. HCMV-specific DNA synthesis was also strongly inhibited by papaverine and DHPG.

• 한국정보통신학회:학술대회논문집
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• 한국해양정보통신학회 2010년도 추계학술대회
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• pp.660-662
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• 2010

To control replication of human cytomegalovirus (HCMV) effectively, inhibitors of peptide nucleic acids (PNA) with a gene delivery agent, PEI (polyethylenimine) against HCMV were applied. The transfection of these PNA inhibitors with PEI agent into host cells showed synergic inhibition effect of HCMV replication. These inhibition effect was confirmed by methods of RT-PCR, CPE, real-time-PCR, and Western blot.

• 미생물학회지
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• 제37권2호
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• pp.145-150
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• 2001

Human cytomegalovirus (HCMV)의 잠복감염으로 부터의 재활성환느 면역기능이 저하된 사람에게 높은 치사율을 가져오며, 재활성화가 기전의 규명은 매우 중요한 연구과제의 하나이다. HCMV의 잠복감염 부위 중 하나라고 생각되고 있는 인체단핵세포에 대한 HCMV의 영향을 알아보기 위해 여러 분화 단계에 있는 THP-1과 HL-60 세포에 HCMV를 감염시킨 후 생존 세포의 수와 형태적 변화를 살펴보았다. HL-60 세포가 HCMV 감염에 의해 세포 생존이나 형태에 큰 영향을 받지 않는 반면, 좀 더 분화된 세포인 THP-1은 HCMV 감염에 의해 생존 세포의 수가 감소하였고, 형태적 변화도 나타났다. 이러한 형태적 변화는 세포의 응집력의 증가에 의한 것으로 HCMV 감염에 따른 THP-1 세포 표면의 CD11b 발현 증가와 밀접한 관계가 있는 것으로 생각된다. THP-1 세포에 HCMV를 감염시킨 후 잠복감염이 이루어진 것을 확인하고, 세포 분화 유도제인 TPA와 hydrocortisone을 처리하였을 경우 방러스가 재활성화하여 증식하는지 알아보았다. 바이러스 감염 2일째에 분화시킨 THP-1 세포에서는 분화 5일째부터 다량의 바이러스가 검출되었고, 감염 17일째 분화시킨 세포에서는 분화 후 15일째부터 바이러스가 검출됨을 관찰하였다. 이는 HCMV를 THP-1 세포에 감염 후 분화를 시키면 잠복감염해 있는 HCMV가 재활성화 되는 것이라 생각되고, 감복해 있던 기간이 길어질수록 바이러스의 재활성화정도는 지연된다는 것을 의미한다.

• Journal of Microbiology
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• 제40권3호
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• pp.224-229
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• 2002

The effect of human cytomegalovirus (HCMV) on three human monocyte cell lines at different stages of differentiation was investigated. While the viability of HL-60 cells or U-937 cells was not significantly affected by HCMV infection, the viability of THP-1 cells was reduced. Acridine orange/ethidiurn bromide staining revealed that the reduction of THP-1 cell viability was due to increased apoptotic death following HCMV infection. Apoptosis in HL-60 cells was not affected by HCMV infection, and induction of apoptosis of U-937 cells by HCMV was intermediate between HL-60 and THP-1 cells. Since HL-60 cells are the least differentiated and THP-1 cells are the most differentiated, the induction of apoptosis of human monocytes appears to be related to the degree of cell differentiation. Flow cytometric and confocal microscopic studies using fluorescent calcium indicator Fluo-3 suggested a significant increase in intracellular free calcium concentration (［Ca$\^$2+/］i) in THP-1 cells undergoing apoptosis by HCMV infection. Again ［Ca$\^$2+/］i in HCMV-infected HL-60 cells was not critically altered, and that in HCMV-infected U-937 cells was intermediate between THP-1 cells and HL-60 cells. Calcium influx blockers such as verapamil and nifedipine partially reversed HCMV-induced apoptosis in THP-1 cells.

• Asian Pacific Journal of Cancer Prevention
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• 제15권3호
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• pp.1453-1457
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• 2014

Background: The association of colorectal cancer with human cytomegalovirus (HCMV) is a controversial issue in cancer research. This study aimed to identify the HCMV virus in colorectal cancer tissues and to investigate the association of HCMV with colorectal cancer. In this study, 50 cancer tissue samples and 50 samples without colon cancer were studied in order to identify the HCMV virus through nested-polymerase chain reaction. The virus was identified in 15 cases of colorectal cancer tissues (15/50) and in 5 cases of normal tissues (5/50). Eight cases of adenocarcinoma tissues were in a moderately differentiated stage, and 7 cases had well-differentiated stage tissues that were positive for viral DNA. The findings were statistically evaluated at a significance level of p<0.05. The HCMV virus could playa role in creating malignancy and the progress of cancer through the process of oncomodulation.

• Parasites, Hosts and Diseases
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• 제30권1호
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• pp.53-58
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• 1992

환자는 54세 광부로 1991년 1월 낙반사고로 인해 가벼운 골반 골절상을 받았었고, 골절상은 완치되었으나 설사가 지속하고 체중이 심하게 감소하였다. 발병 약 9 개월만에 시행한 대변검사 및 조직 검사에서 분선충중(strongyloidiasis), 포자충중(isosporiasis) 및 인형 세포거대바이러스(heman cytomegalovirus)에 감염된 사실이 확인되었다. Albendazole로 치료하였으나 약 2개월 후 근로복지공사 장성병원에서 난치 상환 하에 퇴원하였으며, 퇴원 1주 후 사망하였다.

• Journal of Microbiology
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• 제35권2호
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• pp.152-157
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• 1997

Infection of human fibroblast (HF) cells with human cytomegalovirus (HCMV) result in changes in the intracellular level of second messengers. Since nitric oxide (NO) production has been known to be related with other second messengers, it is probable that HCMV infection of HF cells may involve NO. To test this possibility, the amount of NO was measured following ogenous addition of NO generators such as sodium nitroprusside (SNP) or S-nitroso-N-a-cetylpenicillamine (SNAP) immediately after HCMV infection, however, inhibited virus multiplication. Furthermore, immunoblot experiment using monoclonal antibody to HCMV major immediate early (MIE) proteins or CAT assay using pCMVIE/CAT (plasmid containing CAT gene driven by HCMV MIE promoter) revealed that SNP or SNAP blocked the MIE gene expression. SNP was more effective than SNAP in hibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP in inhibiting HCMV multiplication or MIE gene expression. SNP produced more NO than SNAP. Although the mechanism for the inhibition of HCMV multiplication and MIE gene expression by NO is still elusive some correlation with NO-mediated inhibition of HCMV-induced increase in cytosolic free Ca$\^$2+/ concentration ([Ca$\^$2+/]) was observed. The increase of [Ca$\^$2+/] following HCMV infection was inhibited by SNP, and less effectively by SNAP. Raising [Ca$\^$2+/ with bromo-A23187 partially reversed the SNP block of MIE gene expression. Thus, there appear to e some relationships among NO. [Ca$\^$2+/], and HCMV MIE gene expression.

• Journal of Microbiology
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• 제38권4호
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• pp.255-259
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• 2000

For rapid and sensitive measurement of antiviral activities, application of a recombinant virus containing firefly luciferase gene was attempted. Recombinant human cytomegalovirus (HCMV) containing luciferase gene driven by HCMV late gene pp28 promoter (HCMV/pp28-luc) was used to test the antiviral activities of three known compounds and the result was compared with results from the conventional plaque assay for measuring the production of infectious viruses. When human fibroblast cells were infected with HCMV/pp28-luc, luciferase activity was observed at 2 days after infection and reached maximum at 6 days after infection, whereas the production of infectious virus was maximal at 4 days after infection. The antiviral activities of ganciclovir, acyclovir, and papaverine were measured in HFF cells infected with HCMV/PP28-luc and the luciferase activity was compared with the infectious virus titers. Luciferase activity decreased as the concentration of ganciclovir or papaverine increased, while there was a slight decrease in luciferase activity with acyclovir. The level of the decrease in Luciferase activity was comparable to the level of decrease in the production of infectious virus. Therefore, the antiviral assay using recombinant virus HCMV/pp28-luc resulted in sensitivity similar to the conventional plaque assay with a significant reduction in assay time.

• 대한바이러스학회지
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• 제29권4호
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• pp.261-269
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• 1999

The relationship between second messenger cGMP and human cytomegalovirus (HCMV) replication was investigated. First, the intracellular level of cGMP ([cGMP]i) in HCMV-infected cells was measured. The [cGMP]i increased at early times after HCMV infection, reached maximum level at 12 hr and returned to basal level at 24 hr after virus infection, while [cGMP]i in mock-infected cells remained relatively unchanged. Increasing [cGMP]i resulted in enhanced transcription of HCMV major immediate early gene. For early gene expression, cGMP had varying effect. Expression of 1.2 kb RNA decreased and 2.2 kb RNA increased with increasing cGMP, while 2.7 kb RNA gene expression was not affected. HCMV early genes are regulated by immediate early gene, and the effect of cGMP on the regulatory effect of major immediate early gene on early genes was investigated. In the absence of cGMP, major immediate early gene repressed 2.7 kb RNA gene expression, while 1.2 kb RNA and 2.2 kb RNA early genes were not significantly affected. In the presence of $1\;{\mu}M$ cGMP, however, major immediate early gene stimulated the expression of three early genes.