Cells were harvested after 24 h of cytokine treatment. This result confirmed the interaction between VKORC1v2 and CatD. Significance was determined by two-tailed, unpaired Student’s t test with a confidence level of 95%. For these reasons, quantitation of cell-free HHV-8 DNA may be an accurate marker in determining active HHV-8 replication. There is a similar limited amount of in vivo data assessing T cells and DC in PEL and MCD. a) The cells that stained positive for HHV‐8 were predominantly endothelial cells and lymphocytes located within and around the plexiform lesion (large arrow). 3C).
These findings suggested that clone 1A encoded an HHV-8- specific gene product. 4C). LIPS also detected two more positive samples than the ELISA (94% sensitivity and 100% specificity), but ROC analysis of LIPS and ELISA showed no significant difference in diagnostic performance. To check the specificity of the mixed-antigen ELISA, the reactivity and titers of the sera used in a previous experiment were tested again by IFA and Western blot analysis using HHV8-positive cell lines (TY-1 and BCBL-1; data not shown). T1.1 encodes a polyadenylated nuclear RNA of unknown function, which is highly transcribed during the HHV-8 lytic cycle.27 28 45 Figure 4A, top, shows that T1.1 could be readily detected by Northern blot hybridization, indicating that lytic RNAs were more rapidly and effectively transcribed following HHV-8 infection of fMVDECs than had been demonstrated. Total RNA was isolated from untreated (−) or treated (+) cells (lanes 2 to 5) and subjected to one round of poly(A) selection, using type 3 oligo(dT)-cellulose (lanes 6 to 9). PCR analyses showed that 3 of 60 placenta samples harboured viral DNA and immunohistochemical studies performed on two of them revealed the presence of LANA expression in a few endothelial and trophoblast cells (Figure 8).
GR3 cells were transfected with Tat101 DNA or a control vector as described in Materials and Methods. These data are summarized in Table 1. However, certain KSHV immunoregulatory and growth-promoting genes, including vIL-6, vMIR3 and vMIR5, could be activated by Notch signalling independently of the lytic transactivator RTA73. These clinical features are similar to those of infections caused by other members of the herpesvirus family, such as HHV-6 and hCMV. HUVEC monolayers were infected with the purified viral suspension for 3 hours at 37°C. Low socioeconomic status was a contributing risk that may reflect more crowded and less hygienic living conditions as seen in childhood transmission of other oncogenic viruses such as EBV and hepatitis B [6, 15, 54, 55]. The disease may take on a pattern of exacerbations with subsequent spontaneous remissions, whereas in others, a severe acute illness may occur with a rapid downhill course.
Antibodies, Western blotting, and coprecipitations.Antiserum to vIL-6, generated in this laboratory, has been described previously (24). CHO-B2 cells transfected with pCDNA3 (CHO-B2/pCDNA) and human α3 integrin cDNA containing pCDNA3 (CHO-B2 clone B3; gifts from J. We next determined levels of 16 cytokines, chemokines, and growth factors that have been related to KS, in B cell supernatants by cytokine bead array (CBA), i.e., gamma interferon (IFN-γ), IL-1β, -2, -4, -6, -7, -8, -10, and -12, tumor necrosis factor alpha (TNF-α), monocyte chemoattractant protein 1 (MCP-1), MIP-1α (or CCL3), MIP-1β (or CCL4), the regulated on activation, normal T cell expressed and secreted protein (RANTES, or CCL5), IFN-inducible protein 10, and VEGF (data not shown). These results suggest that heart transplant recipients are, overall, epidemiologically similar to kidney transplant recipients and that the pathogenesis of KS and its relation to HHV-8 are very similar in both transplantation types. The same viral subtype, subtype A, was identified in the early kidney allograft tissue and in bone marrow, gastric lymphoma, and KS biopsy specimens. (B) Importance of gp130 in HHV-8 replication in JSC-1 cells. It has been reported that this test is specific and sensitive when compared to results obtained from other assays and with the presence of KS (14).
The patient has remained asymptomatic and in complete remission, and neither Kaposi sarcoma nor lymphoproliferative disease has been diagnosed in the posttransplant period, after a follow-up of approximately 2 years. (D) KSHV LANA (ORF-73) expression in KS. pcDNA3/Rta contained a 3.1-kb genomic sequence encoding Rta, whose expression was driven by the cytomegalovirus immediate-early promoter/enhancer in the vector. Our data do not address the risk of asymptomatically-infected persons developing HHV-8-associated diseases, nor compare shedding patterns between persons with HHV-8 related diseases and asymptomatic infection; the understanding of viral shedding patterns gained from our work may be used to design longitudinal studies to address questions of HHV-8 transmission and disease association. Human herpesvirus 8 (HHV-8; Kaposi’s sarcoma herpesvirus) is consistently found in all clinical forms of Kaposi’s sarcoma (2, 6), AIDS-associated body cavity-based lymphoma or pleural effusion lymphoma (5), and multicentric Castleman’s disease (15, 42). A 36-year-old man from the Comoro Islands was hospitalized at Avicenne Hospital (Bobigny, France) for fever, chills, and weight loss of 1 month’s duration.