Conclusion HIV-1-related inflammatory cytokines are responsible for the induction of KSHV lytic cycle replication in HUVEC and the induction may be, at least in part, mediated by Rta promoter of KSHV.
Results Recombinant cytokines, such as gamma interferon(IFN-γ), hepatocyte growth factor/scatter factor (HGF/SF), and oncostatin M(OSM) not only induce KSHV lytic cycle replication, but also stimulate Rta promoter activity in latently infected HUVECs.
BCBL-1 cells derived from primary effusion lymphoma (PEL) were fused with human herpesvirus 6 (HHV6)-infected JJhan cells to create heterokaryons and subsequently harvested at the different time points after fusion.
The labeled mRNAs were then used as probes in Southern hybridization reactions to identify restriction fragments in the phage genome where new transcripts were initiated during progression of the phage lytic cycle.
The most commonly studied gene, beige (Bg), has been found to block a postrecognition event in the lytic cycle.
Furthermore, the induction of the HHV-8 lytic cycle was not observed under the staurosporine treatment.
The S-adenosylhomocysteine hydrolase (SAH) and 14-3-3 ζ/phospholipase A2 (PLA2) are transcriptionally activated in parallel to the induction of the Epstein-Barr virus (EBV) lytic cycle by the ganglioside IV3NeuAc-nLcOse4Cer.
Expression of EA-D, BZLF1, and BHRF1 was increased in response to both, SAH- and 14-3-3 ζ/PLA2 overexpression indicating the initiation of the EBV lytic cycle.
BCBL-1 cells derived from primary effusion lymphoma (PEL) were fused with human herpesvirus 6 (HHV6)-infected JJhan cells to create heterokaryons and subsequently harvested at the different time points after fusion. Meanwhile, BCBL-1 cells were cultured with HHV6-infected JJhan cells by direct cell-cell contact and collected at the different time points. Furthermore, BCBL-1 cells were cultured in the conditioned medium that were the normal and heat-inactivatd supernatants of HHV6-infected JJhan cells and also...
BCBL-1 cells derived from primary effusion lymphoma (PEL) were fused with human herpesvirus 6 (HHV6)-infected JJhan cells to create heterokaryons and subsequently harvested at the different time points after fusion. Meanwhile, BCBL-1 cells were cultured with HHV6-infected JJhan cells by direct cell-cell contact and collected at the different time points. Furthermore, BCBL-1 cells were cultured in the conditioned medium that were the normal and heat-inactivatd supernatants of HHV6-infected JJhan cells and also collected at the different time points. Finally, HHV6 viral particles obtained were purified by centrifugation and further be used to infect BCBL-1 cells.Simultaneously, BCBL-1 cells were infected with heat-inactivated or UV-irradiated HHV6 viral particles as control. RT-PCR and/or Real-time quantitative PCR were carried out to evaluate the expression of ORF26 mRNA(encoding the minor capsid protein)of Kaposi's sarcoma-associated herpesvirus(KSHV)in above collected BCBL-1 cells. The results were shown as follows: ①Cytopathogenic effect was observed 15h after cell fusion.ORF26 mRNA expression in BCBL-1 cells detected with RT-PCR increased as significantly compared to that of the control.The results from Real-time PCR analysis at different time points demonstrated that ORF26 mRNA expression in BCBL-1 cells fused with HHV6-infected JJhan cells increased more than 2.3-fold when compared with that in BCBL-1 cells fused with normal JJhan cells. ②ORF26 mRNA expression in BCBL-1 cells cocultured with HHV6-infected JJhan cells increased 1.8-fold at 72h as compared to that of BCBL-1 cells cocul-tured with uninfected JJhan cells.Expression of lytic cycle protein K8.1 in BCBL-1 cells cocultured with HHV6-infected JJhan cells increased 2.46-fold at the 5th day as compared to that of the control. ③ORF26 mRNA expression in BCBL-1 cells cultured for 96 hours in conditioned medium whinh came from the normal and heat-inactivated supernatants of HHV6-infected JJhan cells increased 2.73 and 2.22-fold respectively as compared to that of the relative controls. ④Both alive and inactivated HHV6 viral particles may enhance ORF26 mRNA transcription of KSHV in BCBL-1 cells.These results suggested that HHV6 infection could activate KSHV lytic cycle replication.
Objective To determine whether HIV-1-related inflammatory cytokines can activate lytic cycle replication of Kaposi′s sarcoma-associated herpesvirus(KSHV) in human umbilical vein endothelial cells(HUVEC) and to investigate its potential role in Rta promoter of KSHV involved in reactivation. Methods Using an in vitro model system, we examined the effect of recombinant human cytokines that were similar to that produced by HIV-1-infected T cells on KSHV replication in latently infected HUVEC. KSHV reactivation was...
Objective To determine whether HIV-1-related inflammatory cytokines can activate lytic cycle replication of Kaposi′s sarcoma-associated herpesvirus(KSHV) in human umbilical vein endothelial cells(HUVEC) and to investigate its potential role in Rta promoter of KSHV involved in reactivation. Methods Using an in vitro model system, we examined the effect of recombinant human cytokines that were similar to that produced by HIV-1-infected T cells on KSHV replication in latently infected HUVEC. KSHV reactivation was analyzed with Northern blot analysis and quantitative PCR for ORF26 mRNA expression, a gene encoding the KSHV minor capsid protein was only activated during reactivation. The results were extended and confirmed using a luciferase reporter construct driven by the KSHV Rta(replication and transcription activator) promoter, the first promoter activated during KSHV replication. Results Recombinant cytokines, such as gamma interferon(IFN-γ), hepatocyte growth factor/scatter factor (HGF/SF), and oncostatin M(OSM) not only induce KSHV lytic cycle replication, but also stimulate Rta promoter activity in latently infected HUVECs. Conclusion HIV-1-related inflammatory cytokines are responsible for the induction of KSHV lytic cycle replication in HUVEC and the induction may be, at least in part, mediated by Rta promoter of KSHV.
溶解性周期
lytic cycle
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