Development of novel methods for diagnosis of cytomegalovirus and HHV-6 reactivation.

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To develop the sensitive and specific method for diagnosis of human cytomegalovirus (HCMV) and human herpesvinas 6 (HHV-6) reactivation, we have investigated their latency-associated transcripts that we have previously identified. As a result, we have revealed that the latency-associated transcripts of human herpesvinis 6 (H6LTs) were maximally expressed at a fairly stable intermediate stage between latency and reactivation both in vivo and in vitro., and that H6LTs functioned as sources of immediate-early (IE) protein 1 at this stage, which up-regulated the viral reactivation. We have also analyzed the function of HHV-6 IE2 protein that is encoded in the H6LT. We used yeast two-hybrid screening, and found heterogeneous nuclear ribonucleoprotein K (hnRNP K) and the beta subunit of casein kinase 2 (CK2β) specifically interacted with HHV-6 IE2. These findings indicate that the HHV-6 IE2 protein encoded in the H6LT may affect viral and cellular RNA transcription and translation in viral l … More atency and reactivation.To analyze the gene regulation of H6LTs, we have established the recombination system of HHV-6. The recombinant HHV-6 had an enhanced green fluorescent protein-puromycin gene cassette containing the cytomegalovirus major immediate-early promoter. During HHV-6 latency, the cytomegalovinas promoter used the transcription start sites employed in cytomegalovinis latency, which suggests that the transcriptional control of HHV-6 latency may share some common mechanism with HCMV latency.To identify the factor(s) of HHV 6 reactivation, we have studied the association with HHV-6 reactivation and the work-induced fatigue in healthy adults. Reactivation of HHV-6 was examined for viral DNA by semi-quantitative PCR method. As a result, 88% (35/40) of healthy adults shed the reactivated HHV-6 in the saliva during the fatigue, and 23% (9/40) shed HHV-6 after holidays (approximately 1 week). The copy number of HHV-6 DNA was also reduced after holidays. These findings suggest that HHV-6 is reactivated during the work-induced fatigue. Less