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  • Cytomegalovirus
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  • 1
    Language: English
    In: Cellular and Molecular Life Sciences, 2011, Vol.68(6), pp.1079-1090
    Description: Human cytomegalovirus (HCMV) is a major pathogen in immunocompromised individuals. Here, non-toxic concentrations of the anti-cancer kinase inhibitor sorafenib were shown to inhibit replication of different HCMV strains (including a ganciclovir-resistant strain) in different cell types. In contrast to established anti-HCMV drugs, sorafenib inhibited HCMV major immediate early promoter activity and HCMV immediate early antigen (IEA) expression. Sorafenib is known to inhibit Raf. Comparison of sorafenib with the MEK inhibitor U0126 suggested that sorafenib inhibits HCMV IEA expression through inhibition of Raf but independently of signaling through the Raf downstream kinase MEK 1/2. In concordance, siRNA-mediated depletion of Raf but not of MEK-reduced IEA expression. In conclusion, sorafenib diminished HCMV replication in clinically relevant concentrations and inhibited HCMV IEA expression, a pathophysiologically relevant event that is not affected by established anti-HCMV drugs. Moreover, we demonstrated for the first time that Raf activation is involved in HCMV IEA expression.
    Keywords: Human cytomegalovirus ; Sorafenib ; Kinase inhibitor ; Raf ; Immediate early antigen ; Cancer chemotherapy ; Oncomodulation ; Antiviral therapy
    ISSN: 1420-682X
    E-ISSN: 1420-9071
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  • 2
    Language: English
    In: Medical Microbiology and Immunology, 2011, Vol.200(1), pp.1-5
    Description: The question whether human cytomegalovirus may affect cancer diseases has been discussed (very controversially) for decades. There are convinced believers and strict opponents of the idea that HCMV might be able to play a role in the course of cancer diseases. In parallel, the number of published reports on the topic is growing. Recently published and presented (Ranganathan P, Clark P, Kuo JS, Salamat S, Kalejta RF. A Survey of Human Cytomegalovirus Genomic Loci Present in Glioblastoma Multiforme Tissue Samples. 35th Annual International Herpes Workshop, Salt Lake City, 2010) data on HCMV detection in glioblastoma tissues and colocalisation of HCMV proteins with cellular proteins known to be relevant for glioblastoma progression motivated us to recapitulate the current state of evidence.
    Keywords: Cytomegalovirus ; Cancer ; Oncomodulation ; Tumour virus ; Glioblastoma ; Neuroblastoma
    ISSN: 0300-8584
    E-ISSN: 1432-1831
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  • 3
    Language: English
    In: Reviews in Infection, July, 2010, Vol.1(3), p.166(6)
    Description: Background: Human cytomegalovirus (HCMV) is the most frequent viral pathogen in bone marrow transplant (BMT) patients, where it causes significant morbidity and mortality. The results of the two most frequently used laboratory assays for the detection of active HCMV infection, the pp65-antigenemia assay and the polymerase chain reaction (PCR), often differ in BMT patients. Within the context of limited financial resources it is necessary to economize routine laboratory diagnostic procedures. To evaluate if the pp65-antigenemia assay can be replaced by HCMV-PCR we retrospectively assessed 988 samples from 253 BMT patients without additional information on the patient's clinical presentation or therapy. Material and Methods: The results of both assay (a commercially available pp65-antigen assay and an in-house quantitative real time HCMV-PCR) were classified into four different quantitative groups: PCR group (PCR-g) 0 = PCR neg. (〈200 copies/ml); PCR-g 1 = 200-999 copies/ml; PCR-g 2 = 1,000- 9,999 copies/ml; PCR-g 3 = 〉9,999 copies/ml; pp65-antigen group (pp65-g) 0 = 0 positive cells/400,000 leucocytes; pp65-g 1 = 1-3 pos. cells; pp65-g 2 = 4-99 pos. cells and pp65-g 3 = 〉99 pos. cells. Results: The mean value of PCR copies rises with the number of pp65-antigen (pp65-Ag) positive cells in the antigenemia assay. However, in the pp65-g 0 the mean value and the range of the PCR copies are higher than in the pp65-g 1 (pp65-g 0: 1,188 copies, range: 0-200,000 copies; pp65-g 1: 554 copies range: 0-12,900). There is a statistically significant correlation between the results of the pp65-Ag-assay and quantitative PCR (spearman-correlation: cumulative: 0.49, p = 0.01; pp65-g 2: 0.38, p= 0.01; pp65-g 3: 0.44, p = 0.01). Regarding PCR groups, there is no correlation with the results of the quantitative antigenemia assay. Using any positive PCR result as the reference, the pp65-Ag-assay showed 59% / 77% / 60% / 76% for sensitivity / specificity/ positive predictive value / negative predictive value, respectively and a positive likelihood ratio of 2.6. These values rose with the number of PCR copies and pp65-positive cells. A highly positive result (pp65-g 3 (〉99 cells) or PCR-g 3 (〉 9,999 copies/ml)) of either test almost certainly excludes the possibility of a negative result of the other test. Conclusion: When the pp65-Ag result is greater than 3 positive cells per 400,000 leucocytes, the pp65-Ag-assay and the plasma PCR parallel each other. However, highly positive PCR results can be obtained in pp65-Ag-assay negative samples. Our results suggest that both assays complement each other and should be used concomitant. Keywords: Cytomegalovirus, HCMV, pp65, antigenemia, PCR
    Keywords: Physiologic Monitoring -- Methods ; Medical Tests -- Research ; Bone Marrow Transplantation -- Patient Outcomes
    ISSN: 1837-6738
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  • 4
    Language: English
    In: Medical Microbiology and Immunology, 2010, Vol.199(1), pp.53-60
    Description: Since the dynamics of transmission of human cytomegalovirus (HCMV) have not been clarified yet, we assessed a possible change in HCMV seroprevalence in Frankfurt am Main, Germany during the past twenty years and tried to detect variables with an impact on epidemiology. Between 1/1/1988 and 10/15/2008, a total of 54443 serum samples were collected for routine diagnostics and analyzed using Enzygnost Anti HCMV-IgG enzyme immunoassay (Siemens/Dade Behring, Marburg, Germany). Two decades, 1/1/1988–12/31/1997 and 1/1/1998–10/15/2008, and several groups (type of health insurance, gender, age, HIV-status) were evaluated to assess changes in seroprevalence. Regarding both decades, the overall age-adjusted prevalence of HIV-negative patients dropped from 63.70% (confidence interval (CI) 95% 63.15–64.25) to 57.25% (CI 95% 56.57–57.93; P  〈 0.0001). Private health insurance (PHI) patients showed significant lower HCMV seroprevalences than members of obligatory health insurances (OHI) in both decades (1988–1997: PHI = 55.79%, OHI = 64.27%; P  〈 0.0001; 1998–1908: PHI = 47.02%, OHI = 58.74%; P  〈 0.0001). Furthermore, comparing the two decades, there was generally a gender-specific statistically significant decrease in HCMV seroprevalence for males (63.54–55.54%) and females (63.83–58.73%) as well as for members of PHI and OHI (PHI males: 57.59% to 47.19%, PHI females 54.10–46.80%; OHI males: 64.00–57.06%, OHI females 64.50–60.11%). Also, while female HIV-positive patients showed significant difference in HCMV seroprevalence between the two decades (83.17 and 87.80%, P  = 0.023), there was no significant difference in male patients with HIV (88.76 and 87.32% in the first and second decade, respectively ( P  = 0.196). The cumulative HCMV prevalence of all HIV-negative patients tested in the past 20 years demonstrates a biphasic, age-related rise of HCMV seroprevalence throughout all age-groups. The seroprevalence of HCMV has declined between 1988–1997 and 1998–2008 in Frankfurt am Main, Germany. The decline varied between different age-groups. HCMV prevalence correlates with the type of health insurance, gender, age, and HIV-status.
    Keywords: Cytomegalovirus ; Seroepidemiology ; HCMV ; Germany ; Prevalence
    ISSN: 0300-8584
    E-ISSN: 1432-1831
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  • 5
    Language: English
    In: Journal of Clinical Virology, 2011, Vol.51(4), pp.285-288
    Description: Human cytomegalovirus (CMV) is considered as the most common cause of congenital infection in humans and the overall burden for the public health system is rather high. About 1/10 of vertically infected newborns present or develop severe signs of cytomegalic inclusion disease (CID), with the classical triad of chorioretinitis, microcephaly and cerebral calcifications. However the most symptomatic cases are detected postnatal and methods of diagnostic virology raised the questions for the gold standard in laboratory screening. The current problems in diagnosis and therapy are outlined in two different cases: An acute primary CMV infection with no clinical signs of illness in both mother and child and a secondary CMV-infection resulting in necrotizing CMV encephalitis in the fetus. Beside virus detection in whole blood samples and other fluids, newly adopted laboratory assays like the destination of CMV-IgG avidity were necessary. Furthermore a serologic screening for pregnant women should be implicated routinely. Passive IgG treatment of the mother was helpful but the ultimate goal in prevention of congenital CMV infection is to develop a vaccine, which would be administered to seronegative women.
    Keywords: Primary Infection ; Reactivation ; Secondary Infection ; Pregnancy ; Ventriculomegaly ; Ascites ; Biology
    ISSN: 1386-6532
    E-ISSN: 1873-5967
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  • 6
    Language: English
    In: Medical Microbiology and Immunology, 2013, Vol.202(1), pp.37-47
    Description: Although several host factors have been identified to influence the course of HCMV infection, it still remains unclear why in AIDS patients without highly active antiretroviral therapy human cytomegalovirus (HCMV) retinitis is one of the most common opportunistic infections, whereas in other immunosuppressed individuals it has a low incidence. It was suggested that HCMV glycoprotein B strains may be suitable as marker for virulence and HCMV retinitis. Moreover, UL144 ORF, a member of the TNF-α receptor superfamily, may play a crucial role in innate defences and adaptive immune response of HCMV infection. Furthermore, sequence analyses of HCMV genes UL128, UL130, and UL131A as major determinants of virus entry and replication in epithelial and other cell types were performed. To evaluate the association of sequence variability of depicted viral genes with HCMV retinitis and in vitro growth properties in retinal pigment epithelial cells (RPE) and human foreskin fibroblasts (HFF), we compared 14 HCMV isolates obtained from vitreous fluid and urine of AIDS patients with clinically proven HCMV retinitis. Isolates were analyzed by PCR cycle sequencing and phylogenetic analysis. In addition, sequences of HCMV strains AF1, U8, U11, VR1814, and its cell culture adapted derivates were included. Sequence analysis of gB yielded three genetic subtypes (gB type 1 (5 isolates), gB type 2 (12 isolates), and gB type 3 (5 Isolates)), whereas sequence analysis of UL144 showed a greater diversity (7 isolates type 1A, 2 isolates type 1C, 7 isolates type 2, and 3 isolates type 3). In contrast, the UL128, UL130, and UL131A genes of all low-passage isolates were highly conserved and showed no preferential clustering. Moreover, in HFF and RPE cells, all of our HCMV isolates replicated efficiently independently of their genetic subtype. In conclusion, beside a possible link between the gB subtype 2 and HCMV retinitis, our study found no direct evidence for a connection between UL144/UL128/UL130/UL131A genotypes and the incidence of HCMV retinitis in AIDS patients.
    Keywords: Cytomegalovirus ; Viral tropism and virulence ; Retinitis ; AIDS
    ISSN: 0300-8584
    E-ISSN: 1432-1831
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  • 7
    Language: English
    In: Medical Microbiology and Immunology, 2009, Vol.198(2), pp.79-81
    Description: The (possible) relationship between HCMV and cancer has been discussed for decades. Detection of viral DNA, mRNA and/or antigens in tumour tissues as well as seroepidemiologic evidence suggested a role of HCMV infection in several human malignancies. However, controversial clinical results from diVerent groups had raised skepticism about a role of HCMV in cancer.
    Keywords: Cytomegalovirus–Physiology ; Cytomegalovirus Infections–Complications ; Humans–Etiology ; Neoplasms–Virology ; Neoplasms–Virology ; Infections ; Viruses ; Cancer ; Pathology;
    ISSN: 0300-8584
    E-ISSN: 1432-1831
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  • 8
    Language: English
    In: The Journal of Infectious Diseases, 1 September 2000, Vol.182(3), pp.643-651
    Description: In fibroblasts, infection with human cytomegalovirus (HCMV) inhibits expression of the extracellular matrix proteins thrombospondin-1 and -2 (TSP-1 and TSP-2). These effects may depend on expression of HCMV immediate-early (IE) genes, which are activated by cellular transcription factor NF-kB. The influence of HCMV infection on TSP-1 and TSP-2 expression and the ability of different antiviral drugs to prevent these cellular changes in permissive cultures of human retinal glial cells were observed. Ganciclovir inhibited only HCMV late antigen (LA) expression, whereas antisense oligonucleotide ISIS 2922 and peptide SN50, inhibitors of HCMV IE expression and NF-kB activity, respectively, inhibited both IE and LA expression. ISIS 2922 and SN50, but not ganciclovir, prevented down-modulation of TSP1 and TSP-2. The results showed that HCMV-induced down-modulation of TSP-1 and TSP2 in retinal glial cells is prevented by inhibition of HCMV IE expression. These findings may be relevant to pathogenesis and treatment of HCMV retinitis.
    Keywords: Biological sciences -- Biology -- Cytology ; Biological sciences -- Biology -- Microbiology ; Health sciences -- Medical sciences -- Pharmaceutics ; Biological sciences -- Biology -- Microbiology ; Health sciences -- Medical conditions -- Infections ; Biological sciences -- Biochemistry -- Biomolecules ; Physical sciences -- Chemistry -- Chemical compounds ; Biological sciences -- Biology -- Cytology ; Biological sciences -- Biology -- Cytology ; Health sciences -- Medical conditions -- Diseases
    ISSN: 00221899
    E-ISSN: 15376613
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  • 9
    Language: English
    In: Medical Microbiology and Immunology, 2009, Vol.198(4), pp.257-262
    Description: A coupled luminescent method (CLM) based on glyceraldehyde-3-phosphate dehydrogenase released from injured target cells was used to evaluate the cytotoxicity of antigen-specific HLA class I-restricted CTLs. In contrast to established methods, CLM does not require the pretreatment of target cells with radioactive or toxic labeling substances. CTLs from healthy HLA-A2 positive donors were stimulated by autologous dendritic cells (DCs) pulsed with HLA-A2 restricted HCMV-pp65 nonamer peptides. HLA-A2 positive T2 cells or autologous monocytes pulsed with HCMV-pp65 nonamer peptide served as target cells. Lysis was detected only in HCMV-pp65-pulsed target cells incubated with CTLs from seropositive donors stimulated by HCMV-pp65-pulsed DCs. After 3 days, stimulation 38% of T2 cells and 17% of monocytes were lysed at an effector to target ratio of 8:1. In conclusion, CLM represents a highly sensitive, fast, material-saving and non-toxic/non-radioactive method for the measurement of antigen-specific CTL cytotoxic activity.
    Keywords: Cytotoxic T lymphocytes ; HLA-A2-restricted peptide ; Human cytomegalovirus ; Cytotoxicity ; Coupled luminescent method ; Dendritic cells
    ISSN: 0300-8584
    E-ISSN: 1432-1831
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  • 10
    Language: English
    In: Journal of Immunological Methods, 2006, Vol.311(1), pp.164-173
    Description: The detection and quantification of specific T lymphocytes against human cytomegalovirus (HCMV) has proven an important laboratory marker in the monitoring of patients after stem cell transplantation (SCT). In these patients HCMV infections may cause severe disease and death. However, the determination of HCMV-specific T lymphocytes may be limited by lymphopenia occurring after transplantation. We evaluated a commercial test kit for the reliable determination of HCMV-specific T lymphocyte development in lymphopenic patients after stem cell transplantation. Using a whole blood protocol for the flow cytometric detection of antigen-specific CD4 T-helper and CD8 cytotoxic T lymphocytes this test kit measures intracellular cytokine production after stimulation with HCMV antigen. The measurement of HCMV-specific T lymphocytes was feasible when at least 3000 CD4 or 1000 CD8 T cells could be counted by flow cytometry. Detection of HCMV-specific T lymphocytes was possible, on average, 67 (SD ± 61) days after transplantation for CD4 cells and 27 (SD ± 13) days for CD8 cells, thus being still within the critical time for HCMV reactivation. In conclusion, the use of modern test kits permits the measurement of HCMV-specific T lymphocytes in stem cell transplant recipients and may be included in the HCMV monitoring system after SCT.
    Keywords: Flow Cytometric Analysis ; Antigen-Specific T Lymphocytes ; Intracellular Cytokine Expression ; Stem Cell Transplantation ; Human Cytomegalovirus ; Medicine ; Biology
    ISSN: 0022-1759
    E-ISSN: 1872-7905
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