3A and 4B). Increased persistent replication has serious consequences for the host, including severe elastic artery vasculitis (9, 10, 61) and high levels of reactivating cells (17). 6E and F). In the work described here, we have defined an MHV68 ncRNA that plays an essential role in virus dissemination and subsequent peripheral latency establishment. The inference of an N-terminal extension is consistent with the observation that substantial deletions in this portion of VP19c do not abrogate HSV-1 capsid assembly (39). 6A, left).
4C). Human Kaposi’s sarcoma lesion was analyzed by H&E staining (A) and immunohistochemistry staining with antibodies against the latent nuclear antigen LANA (B), markers for endothelial cells CD31 (C), proliferating cells Ki67 (D), T cells CD3 (E), antigen-presenting cells CD80 (F), macrophage IBA-1 (G) and dendritic cells CD11c (H). This suggests that ORF 73 was transcribed without a requirement for de novo viral proteins and continued to be transcribed in the absence of a switch to β-class gene expression. This study provides the first evidence of RTA’s ability to regulate the balance between latency and lytic replication in vivo. It was surprising to note that BMT mice have equivalent latent viral loads after having increased lytic virus; however, our results are consistent with previous studies that demonstrated that lytic virus titers do not necessarily correlate with latent levels of MHV-68.32,33 We hypothesize that much of the lytic replication at day 7 may occur in cell types that do not remain resident in the lung by day 21. Ridker, P.M., Cushman, M., Stamfer, M.J., Tracy, R.P.
As shown in Fig. The 2.8-kb RNA was detected at 4 h and disappeared at 24 h postinfection, and the 2.9-kb RNA was not visible until 8 h, and continued to accumulate until 13 h postinfection. MHV-68-cloned BAC DNA (pMHV-68; 50 ng) was also transfected as a control. We infected BHK-21 cells at a multiplicity of infection of 5 and harvested RNA at 2 and 4 h postinfection for array analysis (data not shown). The results produced from Southern hybridization matched those predicted by sequence analysis (see Results). These results demonstrate that developing B cells in the bone marrow harbor viral genome throughout chronic infection.
Mapping of the 5′ end of the ORF52 transcript by S1 nuclease protection assay. 2A). Given the success of M2 vaccination in reducing viral latency early in the infection, vaccination with latency-associated antigens that are expressed for longer periods in infected cells may cause a more prolonged reduction in latent viral load. Lymphocytes were then extracted from uninfected mice and diluted to 2 × 106 cells in fresh, precooled RPMI medium and exposed to 0.1 copy per cell of either vgp150Δ or vgp150R from lung preparations. Disrupted cells were plated in a similar series of twofold dilutions. Several host proteins are notable among those specifically induced in or lost from infected cells.
(A) Restriction endonuclease profiles of MHV-68 and MHV-76 virion DNAs. Bone marrow-derived macrophages from control (BL6) and IRF-1−/− mice were infected with MHV68 at an MOI of 5 (A) or 0.01 (B) PFU/cell. While infection of SAP-deficient mice with MHV68 mimics several features of SAP deficiency in humans, XLP is not fully recapitulated , , . In addition to the aforementioned M1Δ511 mutant and control virus, a panel of individual viral mutants was compiled consisting of three recombinant viruses harboring translation stop codons within each of the M1 neighboring genes (M2, M3, and M4), as well as a large-scale deletion mutant (Δ9473) that spans nearly 9.5 kb and encompasses the entire left end of the genome, including M1 () (17–19, 22). Mice, infections, and organ harvest.p18INK4c−/− (gene name, CDKN2C) and p27Kip1−/− (gene name, CDKN1B) mice on a C57BL/6 background (David Franklin, Purdue University) were previously described (25). On the other hand, M2 expression leads to inhibition of AKT activation upon BCR stimulation.
Liu, and S. The association of viral genomes with PML NBs has also been noted with cytomegalovirus (CMV), varicella-zoster virus, Epstein-Barr virus (EBV), and other DNA viruses including adenoviruses, simian virus 40, polyomavirus, and the parvovirus adeno-associated virus (22). Thus, we sought to determine whether there were basal differences in the ability of the AECs from Balb/c or TLR-9-/- mice to control fibroblast proliferation.