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Berlin Brandenburg

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  • 1
    Language: English
    In: Viral immunology, December 2011, Vol.24(6), pp.455-61
    Description: Cytomegalovirus(CMV) reactivation causes immunopathy, graft malfunction, and even rejection. The traditional anti-CMV drug ganciclovir is not able to prevent reactivation of endogenous virus. Recent studies have found that proteasome inhibitor (PI) is able to suppress CMV replication. In this study we investigated the influence of proteasome inhibitor MG132 and ganciclovir on the CMV-specific CD8(+) T-cell immune response. We found that interferon-γ (IFN-γ) production in response to CMV-infected fibroblasts was reduced under the influence of MG132 in a dose-dependent manner. A marked reduction was observed at 0.5 μM. Likewise, CMV-specific cytotoxicity of CD8(+) T cells was decreased in the presence of MG132. In contrast, the traditional CMV replication inhibitor ganciclovir (10 μM) had no such effect. These findings might have important implications in reducing CMV-associated immunopathy by altering epitope generation through the application of selective proteasome inhibitors.
    Keywords: Immunity, Cellular ; Antiviral Agents -- Pharmacology ; Cd8-Positive T-Lymphocytes -- Drug Effects ; Cysteine Proteinase Inhibitors -- Pharmacology ; Ganciclovir -- Pharmacology ; Leupeptins -- Pharmacology
    ISSN: 08828245
    E-ISSN: 1557-8976
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  • 2
    Language: English
    In: Journal of clinical microbiology, May 2004, Vol.42(5), pp.2298-300
    Description: Due to a paucity of published data concerning the prevalence of viral nucleic acid in homografts, we analyzed tissue from 30 donor hearts for the presence of viral genome sequences of enteroviruses, adenoviruses, human cytomegalovirus, and influenza virus using different PCR techniques. Viral DNA was amplified in 64 and 52% of the subvalvular myocardial tissue and non-coronary valve samples, respectively. These findings, compared with clinical history and histologic and serologic analysis, demonstrate the importance of viral safety measures in heart valve banking.
    Keywords: Bioprosthesis -- Virology ; DNA, Viral -- Isolation & Purification ; Heart Valve Prosthesis -- Virology
    ISSN: 0095-1137
    E-ISSN: 1098660X
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  • 3
    Language: English
    In: The Journal of biological chemistry, 21 February 2003, Vol.278(8), pp.5597-604
    Description: The molecular mechanisms underlying the regulation of interleukin (IL)-10 transcription in monocytic cells by various stimuli during inflammation and the stress reaction are not fully understood. Recently, we provided evidence that stress-induced IL-10 promoter activation in monocytic cells is mediated by catecholamines via a cAMP-dependent signaling pathway including CREB/ATF (cAMP-responsive element binding protein/activating transcription factor) binding to two CRE motifs. However, the mutation of these sites diminished cAMP responsiveness by only 50%, suggesting a role for additional transcription factors and elements in the cAMP-dependent regulation of the human IL-10 promoter. Here, we analyze the functional role of one such factor, C/EBP, in two cell lines of myelomonocytic origin, THP-1 and HL-60, which are known to differ in their differentiation status and C/EBP protein content. We show that the level of basal as well as cAMP-stimulated IL-10 transcription depends on the expression of C/EBP alpha and beta and their binding to three motifs in the promoter/enhancer region. The C/EBP5 motif, which is located between the TATA-box and the translation start point, is essential for the C/EBP-mediated constitutive and most of the cAMP-stimulated expression as its mutation nearly abolished IL-10 promoter activity. Our results suggest a dominant role of C/EBP transcription factors relative to CREB/ATF in tissue-specific and differentiation-dependent IL-10 transcription.
    Keywords: Promoter Regions, Genetic ; Bucladesine -- Pharmacology ; Ccaat-Enhancer-Binding Proteins -- Genetics ; Cyclic Amp -- Physiology ; Gene Expression Regulation -- Immunology ; Interleukin-10 -- Genetics ; Monocytes -- Cytology ; Transcription, Genetic -- Immunology
    ISSN: 0021-9258
    E-ISSN: 1083351X
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  • 4
    Language: English
    In: Journal of Biological Chemistry, 11/02/2001, Vol.276(44), pp.40712-40720
    Description: Recently we demonstrated that the ability of tumor necrosis factor alpha (TNF alpha ) to stimulate the human cytomegalovirus (HCMV) IE1/2 enhancer/promoter activity in myeloid progenitor-like cells decreases when these cells differentiate into promonocytic cells. In addition, TNF alpha stimulation in the progenitor-like cell line HL-60 was shown to be mediated by nuclear factor Kappa B (NF- Kappa B) activation and its binding to the 18-base pair sequence motifs of the IE1/2 enhancer. We demonstrate here that the cell differentiation-dependent reduction of TNF alpha stimulation is not due to insufficient NF- Kappa B activation but correlates with increased synthesis of the monocyte differentiation- associated factors CCAAT/enhancer-binding protein (C/EBP) alpha and beta . Overexpression of C/EBP alpha / beta in HL-60 cells, which normally produce only very small amounts of C/EBP, stimulated the basal activity of the promoter in the absence of NF- Kappa B but suppressed the stimulatory effect of TNF alpha . A novel C/EBP-binding site was identified in the IE1/2 enhancer directly downstream of a NF- Kappa B site. In order to understand the mechanisms of interaction, we used an IE1/2 promoter mutant that failed to bind C/EBP at this position and several constructs that contained exclusively NF- Kappa B- and/or C/EBP-binding sites upstream of the minimal IE1/2 promoter. We could demonstrate that C/EBP alpha / beta interacts with NF- Kappa B p65 and displays inhibitory activity even in the absence of direct DNA binding by forming p65-C/EBP-containing protein complexes bound to the NF- Kappa B site. Moreover, C/EBP binding to the DNA adjacent to NF- Kappa B supports the down-regulatory effect of C/EBPs possibly due to stabilization of a multimeric NF- Kappa B-C/EBP complex. Our results show that cell differentiation factors may interfere with TNF alpha -induced human cytomegalovirus gene (re)activation.
    Keywords: Human Cytomegalovirus ; Human Cytomegalovirus ; Human Cytomegalovirus ; Nf-B Protein ; Monocytes ; Gene Regulation ; Tumor Necrosis Factor-a ; Nf-^Kb Protein ; Monocytes ; Gene Regulation ; Tumor Necrosis Factor-^a ; Tumor Necrosis Factor-^a ; Transcription Factors ; Myeloid Cells ; C/Ebpa Protein ; C/Ebpb Protein ; Nf-^Kb Protein ; C/Ebp^a/^B Protein ; C/Ebp^a Protein ; C/Ebp^B Protein ; Transcription Factors ; Effects on Host Cell Metabolism ; Viruses ; Man ; C/Ebp Alpha Protein ; C/Ebp Alpha / Beta Protein ; C/Ebp Beta Protein ; Nf- Kappa B Protein ; Man;
    ISSN: 0021-9258
    E-ISSN: 1083-351X
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  • 5
    In: Critical Care Medicine, 2001, Vol.29(9), pp.1786-1791
    Description: OBJECTIVE: Partial liquid ventilation with perfluorocarbons (PFC) might be used as a new ventilatory strategy to treat respiratory insufficiency in congenital pneumonia. The present study investigates for the first time effects of PFC on growth and viability of group B streptococci (GBS) and Escherichia coli, bacteria frequently causing congenital pneumonia. DESIGN: Prospective, in vitro study. SETTING: Research laboratory in a university. MATERIAL: Group B streptococci 090 Ia HD Colindale and E. coli K12, JM 101. INTERVENTIONS: E. coli (10/mL) were grown in the absence or presence of different PFC (RM 101, PF 5080, FO 6167) for up to 6 hrs. To study bacterial viability, GBS (5×10/mL) were incubated in saline with or without different PFC, PFC/surfactant emulsions, or surfactant (Curosurf) for up to 5 hrs. Every 2 hrs, the colony forming units were determined by plating different dilutions of bacteria on agar. MEASUREMENTS AND MAIN RESULTS : RM 101 or PF 5080 alone and in emulsions with surfactant had no effect on viability of GBS or growth of E. coli. For FO 6167, a previously described toxicity was found, even if 1 mL of GBS suspension was incubated with only 100 μL of FO 6167, verifying the experimental design that guarantees a PFC bacteria contact. The toxic effects were almost prevented by forming a PFC-in-surfactant emulsion but not by preincubation of GBS with surfactant and subsequent FO 6167 exposure. CONCLUSION: RM 101 and PF 5080 did not influence bacterial growth in vitro; direct effects on bacterial proliferation during partial liquid ventilation in congenital pneumonia seem, therefore, unlikely. Interestingly, we found that the known toxic effects of FO 6167 can be prevented by covering PFC with a surfactant film. Surfactant reduced the cytotoxic effects of FO 6167, probably by preventing a direct contact between FO 6167 and the bacterial cell wall.
    Keywords: Biological Products–Drug Effects ; Emulsions–Growth & Development ; Escherichia Coli–Pharmacology ; Fluorocarbons–Pharmacology ; Liquid Ventilation–Drug Effects ; Phospholipids–Growth & Development ; Pulmonary Surfactants–Growth & Development ; Streptococcus Agalactiae–Growth & Development ; Abridged ; Biological Products ; Emulsions ; Fluorocarbons ; Phospholipids ; Pulmonary Surfactants ; Poractant Alfa;
    ISSN: 0090-3493
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  • 6
    Language: English
    In: Journal of Clinical Virology, 1999, Vol.12(2), pp.96-96
    Keywords: Biology
    ISSN: 1386-6532
    E-ISSN: 1873-5967
    Source: ScienceDirect Journals (Elsevier)
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  • 7
    Language: English
    In: Experimental dermatology, July 2011, Vol.20(7), pp.561-7
    Description: Previously, we have reported a frequent association of active plaque psoriasis with inflammation-mediated cytomegalovirus (CMV) reactivation. This study aimed at characterizing the impact of CMV infection on psoriasis disease activity and peripheral cellular adaptive immune response. Twenty nine patients with active plaque psoriasis and 29 healthy controls were analysed for CMV-serostatus, CMV-antigenaemia, frequencies of peripheral CMV-specific T cells and the immunophenotype of peripheral CD8+ T cells. (i) Psoriasis severity was higher in CMV-seropositive patients and positively correlated to the severity of CMV-antigenaemia. (ii) In comparison to CMV-seropositive healthy controls, CMV-seropositive psoriasis patients showed a reduced frequency of circulating CMV-specific T cells that increased under effective antipsoriatic therapy. (iii) The immunophenotype of peripheral CD8+ T cells was dominated by CMV-seroprevalence. (iv) Selective analysis of CMV-seronegative psoriasis patients revealed a strong expansion of a - probably early activated - CD8+ T-cell population with the yet undescribed differentiation phenotype 'CD45RA-dim/CD11a-dim'. Under effective antipsoriatic therapy this population decreased in parallel to an increase of effector differentiated CD8+ T cells. Taken together with our previous results of inflammation-mediated CMV reactivation in psoriasis, our data support the concept of an interactive relationship between psoriasis and CMV infection which may be mediated by peripheral CD8+ T cells.
    Keywords: Cd8-Positive T-Lymphocytes -- Immunology ; Cytomegalovirus Infections -- Complications ; Psoriasis -- Complications
    ISSN: 09066705
    E-ISSN: 1600-0625
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  • 8
    Language: English
    In: Science translational medicine, 25 September 2013, Vol.5(204), pp.204ra129
    Description: Psoriasis and atopic dermatitis (AD) are the most common chronic inflammatory skin diseases. Although both patient groups show strongly impaired skin barrier function, only AD patients frequently suffer from cutaneous viral infections. The mechanisms underlying the distinct susceptibilities to these pathogenetic and often life-threatening infections are unknown. We show that antiviral proteins (AVPs) such as MX1, BST2, ISG15, and OAS2 were strongly elevated in psoriatic compared to AD lesions and healthy skin. Of 30 individually quantified cytokines in psoriatic lesions, interleukin-29 (IL-29) was the only mediator whose expression correlated with the AVP levels. IL-29 was absent in AD lesions, and neutralization of IL-29 in psoriatic skin reduced AVP expression. Accordingly, IL-29 raised AVP levels in isolated keratinocytes, epidermis models, and human skin explants, but did not influence antibacterial protein production. AVP induction correlated with increased antiviral defense of IL-29-treated keratinocytes. Furthermore, IL-29 elevated the expression of signaling elements, resulting in increased sensitivity of keratinocytes toward its own action. We identified T helper 17 (T(H)17) cells as IL-29 producers and demonstrated their ability to increase the antiviral competence of keratinocytes in an IL-29-dependent manner. Transforming growth factor-β and the activity of RORγt/RORα were most critical for the development of IL-29-producing T(H)17 cells. IL-29 secretion by these cells was dependent on NFAT and c-Jun N-terminal kinase and was inhibited by IL-4. These data suggest that T(H)17 cell-derived IL-29, which is absent in AD, mediates the robust antiviral state on psoriatic skin, and demonstrate a new function of T(H)17 cells.
    Keywords: Interleukins -- Immunology ; Psoriasis -- Immunology ; Skin -- Immunology ; Th17 Cells -- Immunology
    ISSN: 19466234
    E-ISSN: 1946-6242
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  • 9
    Language: English
    In: Journal of clinical microbiology, March 2005, Vol.43(3), pp.1318-24
    Description: In a clinical trial, the incidence of cytomegalovirus reactivation in breastfeeding mothers and transmission to their preterm infants were studied. Breast milk from 73 mothers as well as urine and tracheal and pharyngeal aspirates from their 89 infants were screened weekly for human cytomegalovirus (HCMV) DNA during the first 2 months after delivery. Of the 73 mothers, 48 (66%) were positive for HCMV DNA in the lactating breast. HCMV reactivation could be confirmed for 19 of 20 (95%) immunoglobulin G-positive mothers. Of the eight immunoglobulin G-negative mothers one was positive for HCMV DNA in breast milk. In only 2 out of 13 seropositive mothers with HCMV DNA in breast milk could viral DNA be detected in the peripheral blood. HCMV mother-to-child transmission was concluded for 20 of the 48 (42%) mothers positive for DNA or 7 of 19 (37%) seropositive for HCMV and positive for HCMV DNA in breast milk and one of one mother seronegative for HCMV but positive for HCMV DNA in breast milk. One mother transmitted the virus to her twins. In addition, one infant acquired postnatal HCMV infection despite the mother's being negative for HCMV DNA in breast milk; altogether, we found 22 infants with HCMV infection. In 13 of these 22 infants, virus infection occurred definitively postnatally; two of them developed severe symptomatic HCMV infection. HCMV-infected infants demonstrated higher incidences of amniotic infection, respiratory distress syndrome, bronchopulmonary dysplasia, and retinopathia praenatalis than noninfected infants, however, the differences were not statistically significant. In summary, our study confirmed a very high incidence of HCMV reactivation in mothers during lactation and a significant risk of transmission to preterm infants with the possibility of severe disease in these babies.
    Keywords: Infectious Disease Transmission, Vertical ; Virus Activation ; Cytomegalovirus -- Physiology ; Cytomegalovirus Infections -- Transmission
    ISSN: 0095-1137
    E-ISSN: 1098660X
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  • 10
    Language: English
    In: Biological chemistry, October 2002, Vol.383(10), pp.1601-9
    Description: From clinical studies the proinflammatory cytokine TNFalpha was proposed to play a key role in human cytomegalovirus (HCMV) reactivation from latency. In vitro experiments confirmed that TNFalpha stimulates the activity of the HCMV IE1/2 enhancer/promoter, which controls immediate early protein IE1 and IE2 gene expression via activation of the transcription factor NF-kappaB and its binding to putative binding sites in the IE1/2 enhancer. NF-kappaB was also proposed to be involved in IE1-mediated autostimulation of this promoter. The IE1/2 enhancer of HCMV contains four putative NF-kappaB binding sites which differ in their distance to the transcription start site as well as in their sequence. Construction and testing of a series of promoter mutants demonstrated that NF-kappaB is essential for both TNFalpha and IE1 stimulation. Furthermore, we were able to show that although all four NF-kappaB sites bind NF-kappaB with similar affinity in vitro, the contribution to TNFalpha and IE1 stimulation differs in correlation with the distance to the transcription start site and the sequence. Site 1 and 3 play the most dominant role and site 2 an intermediate, while site 4, which is conserved in sequence but far distant from the transcription start site, had no influence on NF-kappaB-mediated regulation of the IE1/2 promoter. Specific inhibition of NF-kappaB signalling by co-expression of a dominant-negative IkappaB variant reduced TNFalpha stimulation of the IE1/2 enhancer/promoter by up to 80%. From this data, inhibitors of NF-kappaB activation are suggested to be an alternative therapeutical strategy to interfere with HCMV (re)activation in undifferentiated monocyte/granulocyte progenitor cells in patients with a high risk of inflammation-related HCMV (re)activation.
    Keywords: Viral Proteins ; Cytomegalovirus -- Physiology ; Nf-Kappa B -- Antagonists & Inhibitors
    ISSN: 1431-6730
    E-ISSN: 14374315
    Source: MEDLINE/PubMed (U.S. National Library of Medicine)
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