MAB8131 Sigma-AldrichAnti-Cytomegalovirus Antibody, immediate early, clone 6F8.2

Human cytomegalovirus (HCMV) has been indicated being a significant oncomodulator. Recent reports have suggested that an antiviral treatment alters the outcome of a glioblastoma. We analysed the performance of commercial HCMV-antibodies applying the immunohistochemical (IHC) methods on brain sample obtained from a subject with a verified HCMV infection, on samples obtained from 14 control subjects, and on a tissue microarray block containing cores of various brain tumours. Based on these trials, we selected the best performing antibody and analysed a cohort of 417 extra- and intra-axial brain tumours such as gliomas, medulloblastomas, primary diffuse large B-cell lymphomas, and meningiomas. HCMV protein pp65 immunoreactivity was observed in all types of tumours analysed, and the IHC expression did not depend on the patient's age, gender, tumour type, or grade. The labelling pattern observed in the tumours differed from the labelling pattern observed in the tissue with an active HCMV infection. The HCMV protein was expressed in up to 90% of all the tumours investigated. Our results are in accordance with previous reports regarding the HCMV protein expression in glioblastomas and medulloblastomas. In addition, the HCMV protein expression was seen in primary brain lymphomas, low-grade gliomas, and in meningiomas. Our results indicate that the HCMV protein pp65 expression is common in intra- and extra-axial brain tumours. Thus, the assessment of the HCMV expression in tumours of various origins and pathologically altered tissue in conditions such as inflammation, infection, and even degeneration should certainly be facilitated.

Human cytomegalovirus (HCMV) infection is associated with a series of direct and indirect effects following renal transplantation. However, the presence of HCMV in the kidney and its relationship with acute rejection and long-term graft function remain to be fully elucidated. Sixty-two biopsies derived from 30 renal transplant recipients with signs of clinical rejection were analyzed for HCMV using a sensitive in situ DNA hybridization method. Biopsies were also subjected to staining with anti-C4d antibodies and an anti-caspase 3 antibody to detect humoral rejection and apoptosis, respectively. In 21 patients, serial serum creatinine levels over 5 years of follow-up were analyzed. HCMV DNA was detected in biopsies from 21/30 (70%) of the patients and 32/62 (52%) of the individual biopsies. HCMV DNA was detected early after transplant and was localized to renal tubule epithelial cells but not associated with apoptosis. HCMV DNAemia developed within 2 weeks of detecting HCMV DNA in the biopsy in 53% of patients. Ninety percent of patients experiencing HCMV disease had HCMV DNA in their biopsy. HCMV DNA was equally distributed between patients with or without histological evidence of acute rejection and was detected more frequently in patients with peritubular C4d deposits. Creatinine levels at 12 months post-transplant were significantly higher in patients with HCMV DNA and remained elevated over the 5 years of follow-up. HCMV DNA is frequently detected in renal tubular epithelial cells early after renal transplantation, precedes DNAemia and is associated with poor long-term graft function.

Endothelial cells are natural sites of infection for human cytomegalovirus (HCMV) and are increasingly recognized to play an important role in viral dissemination, as well as provide access to underlying tissues and organs. However, the viral factors required for endothelial cell tropism are poorly defined. The goals of the project were to develop a system to study endothelial cell infectivity factors in HCMV, and map the viral genetic determinants required for these tropism functions. HCMV infection of primary aortic endothelial cells (AEC) was studied as a means to evaluate aspects relevant to both pathogenesis of acute infection and chronic vascular diseases. A series of HCMV virus strains was screened for endothelial tropism by comparing replication efficiencies on AEC. A virus strain that was efficient for replication (AD169varATCC), and a virus strain that was restricted for replication (Toledo), were selected for further analysis and characterization. We present evidence for a novel HCMV endothelial tropism factor that functioned following viral internalization across the endothelial cell plasma membrane and prior to nuclear entry. This factor may be involved in intracellular transport of the virion capsid-tegument structure. Complementation approaches using pseudotype virus infection of AEC demonstrated that the tropism defective strain could be rescued in trans. This supported the existence of a viral encoded tropism determinant. Using a gain of function approach, endothelial cell infectivity of the non-tropic HCMV strain Toledo was rescued with AD169 cosmid sequences. Tropism-specific viral genetic determinant(s) may be mapped to a region of the AD169 viral DNA encompassing UL48-56.