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In Vitro Replication of Varicella-Zoster Virus in Human Retinal Pigment Epithelial Cells (2008)

Schmidt-Chanasit, Jonas ; Bleymehl, Karoline ; Rabenau, Holger F. ; [et al.]

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In: Journal of Clinical Microbiology, 2008, Vol. 46(6), p.2122

Description: Here we describe for the first time the productive in vitro infection of human retinal pigment epithelial cells by varicella-zoster virus (VZV), resulting in a typical cytopathic effect (CPE) that is characterized by enlarged cells with increased granularity. Depending on the CPE dissemination, high titers of up to 1.6 x 10 super(6) PFU of cell-free and cryostable VZV/ml can be recovered.

Keywords: Retinal Pigment Epithelium ; Retina ; Replication ; Pigments ; Infection ; Varicella-Zoster Virus ; Replication;

ISSN: 0095-1137

ISSN: 00951137

E-ISSN: 1098660X

DOI: 10.1128/JCM.00122-08

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2

Article

Retroviral Vectors Pseudotyped with Severe Acute Respiratory Syndrome Coronavirus S Protein (2004)

Giroglou, Tsanan ; Cinatl, Jindrich, Jr. ; Rabenau, Holger ; [et al.]

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In: The Journal of Virology, 2004, Vol. 78(17), p.9007

Description: The worldwide outbreak of severe acute respiratory syndrome (SARS) was shown to be associated with a novel coronavirus (CoV) now called SARS CoV. We report here the generation of SARS CoV S protein-pseudotyped murine leukemia virus (MLV) vector particles. The wild-type S protein pseudotyped MLV vectors, although at a low efficiency. Partial deletion of the cytoplasmic tail of S dramatically increased infectivity of pseudotypes, with titers only two- to threefold lower than those of pseudotypes generated in parallel with the vesicular stomatitis virus G protein. S-pseudotyped MLV particles were used to analyze viral tropism. MLV(SARS) pseudotypes and wild-type SARS CoV displayed similar cell types and tissue and host restrictions, indicating that the expression of a functional receptor is the major restraint in permissiveness to SARS CoV infection. Efficient gene transfer could be detected in Vero and CaCo2 cells, whereas the level of gene marking of 293T, HeLa, and HepG2 cells was only slightly above background levels. A cat cell line and a dog cell line were not susceptible. Interestingly, PK-15, a porcine kidney cell line, and primary porcine kidney cells were also highly permissive for SARS S pseudotypes and wild-type SARS CoV. This finding suggests that swine may be susceptible to SARS infection and may be a source for infection of humans. Taken together, these results indicate that MLV(SARS) pseudotypes are highly valuable for functional studies of viral tropism and entry and, in addition, can be a powerful tool for the development of therapeutic entry inhibitors without posing a biohazard to human beings.

Keywords: Sars Coronavirus ; Murine Leukemia Virus ; Vesicular Stomatitis Virus ; Sars Coronavirus ; Murine Leukemia Virus ; Vesicular Stomatitis Virus ; Expression Vectors ; Infection ; Host Range ; Severe Acute Respiratory Syndrome ; Kidney ; Tropism ; Leukemia ; Guanine Nucleotide-Binding Protein ; Gene Transfer ; Deletion ; Tails ; Infectivity ; Vesicular Stomatitis ; Background Levels ; Expression Vectors ; Severe Acute Respiratory Syndrome ; Kidney ; Tropism ; Guanine Nucleotide-Binding Protein ; Gene Transfer ; Deletion ; Infectivity ; Viral Genetics Including Virus Reactivation ; Cloning Vectors ; S Protein ; S Protein;

ISSN: 0022-538X

ISSN: 0022538X

E-ISSN: 10985514

DOI: 10.1128/JVI.78.17.9007-9015.2004

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3

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Human cytomegalovirus infection in tumor cells of the nervous system is not detectable with standardized pathologico-virological diagnostics (2014)

Baumgarten, Peter ; Michaelis, Martin ; Rothweiler, Florian ; [et al.]

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In: Neuro-Oncology, 2014, Vol. 16(11), pp.1469-1477

Description: BACKGROUND: Experimental findings have suggested that human cytomegalovirus (HCMV) infection of tumor cells may exert oncomodulatory effects that enhance tumor malignancy. However, controversial findings have been published on the presence of HCMV in malignant tumors. Here, we present the first study that systematically investigates HCMV infection in human nervous system tumors by highly sensitive immunohistochemistry in correlation with the HCMV serostatus of the patients.METHODS: Immunohistochemical and quantitative PCR-based methods to detect different HCMV antigens and genomic HCMV DNA were optimized prior to the investigation of pathological samples. Moreover, the pathological results were matched with the HCMV serostatus of the patients.RESULTS: HCMV immediate-early, late, and pp65 antigens could be detected in single cells from HCMV strain Hi91-infected UKF-NB-4 neuroblastoma cells after 1:1024 dilution with noninfected UKF-NB-4 cells. Genomic HCMV DNA could be detected in copy numbers as low as 430 copies/mL. However, we did not detect HCMV in tumors from a cohort of 123 glioblastoma, medulloblastoma, or neuroblastoma patients. Notably, we detected nonspecifically positive staining in tumor tissues of HCMV seropositive and seronegative glioblastoma patients. The HCMV serostatus of 67 glioblastoma patients matched the general epidemiological prevalence data for Western countries (72% of female and 57% of male glioblastoma patients were HCMV seropositive). Median survival was not significantly different in HCMV seropositive versus seronegative glioblastoma patients.CONCLUSIONS: The prevalence of HCMV-infected tumor cells may be much lower than previously reported based on highly sensitive detection methods.

Keywords: Cytomegalovirus ; Glioma ; Oncomodulation

ISSN: 1522-8517

E-ISSN: 1523-5866

DOI: 10.1093/neuonc/nou167

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4

Article

Nano-coating protects biofunctional materials (2012)

Tscheliessnig, Rupert ; Zörnig, Martin ; Herzig, Eva M ; [et al.]

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Language: English

In: Materials Today, September 2012, Vol.15(9), pp.394-404

Description: The demand to develop convergent technology platforms, such as bio-functionalized medical devices, is rapidly increasing. However, the loss of biological function of the effector molecules during sterilization represents a significant and general problem. Therefore, we have developed and characterized a nano-coating (NC) formulation capable of maintaining the functionality of proteins on biological-device combination products. As a proof of concept, the NC preserved the structural and functional integrity of an otherwise highly fragile antibody immobilized on polyurethane during deleterious sterilizing irradiation (≥ 25 kGy). The NC procedure enables straight-forward terminal sterilization of bio-functionalized materials while preserving optimal conditioning of the bioactive surface.

Keywords: Engineering

ISSN: 1369-7021

E-ISSN: 1873-4103

DOI: 10.1016/S1369-7021(12)70166-9

URL: View record in ScienceDirect (Access to full text may be restricted)

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5

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Identification of a Novel Coronavirus in Patients with Severe Acute Respiratory Syndrome (2003)

Drosten, Christian ; Günther, Stephan ; Preiser, Wolfgang ; [et al.]

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In: The New England Journal of Medicine, 2003, Vol.348(20), pp.1967-1976

Description: Background The severe acute respiratory syndrome (SARS) has recently been identified as a new clinical entity. SARS is thought to be caused by an unknown infectious agent. Methods Clinical specimens from patients with SARS were searched for unknown viruses with the use of cell cultures and molecular techniques. Results A novel coronavirus was identified in patients with SARS. The virus was isolated in cell culture, and a sequence 300 nucleotides in length was obtained by a polymerase-chain-reaction (PCR)–based random-amplification procedure. Genetic characterization indicated that the virus is only distantly related to known coronaviruses (identical in 50 to 60 percent of the nucleotide sequence). On the basis of the obtained sequence, conventional and real-time PCR assays for specific and sensitive detection of the novel virus were established. Virus was detected in a variety of clinical specimens from patients with SARS but not in controls. High concentrations of viral RNA of up to 100 million molecules per milliliter were found in sputum. Viral RNA was also detected at extremely low concentrations in plasma during the acute phase and in feces during the late convalescent phase. Infected patients showed seroconversion on the Vero cells in which the virus was isolated. Conclusions The novel coronavirus might have a role in causing SARS. This study used cell culture and molecular techniques to identify the infectious agent associated with SARS. A novel coronavirus was found in multiple samples from 18 patients but in no specimens from control subjects. In the patients there were high concentrations of viral RNA in sputum, a finding consistent with a highly infectious agent. Low concentrations of viral RNA were also detected in stool. The severe acute respiratory syndrome (SARS) was recently identified as a new clinical entity.1,2 Patients present with fever, dry cough, dyspnea, headache, and hypoxemia. Typical laboratory findings are lymphopenia and mildly elevated aminotransferase levels. Death may result from progressive respiratory failure due to alveolar damage.3 SARS appears to be caused by an unknown infectious agent that is transmitted from human to human. The World Health Organization (WHO) had recorded 2353 cases by April 4, 2003. About 4 percent of patients with SARS have died.4 The SARS epidemic started in Asia, with the majority of cases occurring in China and . . .

Keywords: Medicine;

ISSN: 0028-4793

E-ISSN: 1533-4406

DOI: 10.1056/NEJMoa030747

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6

Article

Long-term productive human cytomegalovirus infection of a human neuroblastoma cell line (1996)

Cinatl, Jindrich ; Vogel, Jens-U. ; Cinatl, Jaroslav ; [et al.]

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Language: English

In: International Journal of Cancer, 01/03/1996, Vol.65(1), pp.90-96

Description: Human neuroblastoma cell line UKF-NB-4 persistently infected with human cytomegalovirus (HCMV) strain AD169 was established to study the effects of long-term HCMV infection on virus production and phenotypic characteristics of tumour cells. The cells designated UKF-NB-4 super(AD169) were subcultured (80 subcultures) over a period of more than 2 years after initiation of infection. UKF-NB-4 super(AD169) cells continued to produce infectious virus in successive passages, with a titre ranging from 9 x 10 super(3) to 1 x 10 super(5) and from 2 x 10 super(1) to 2 x 10 super(2) plaque-forming units per 10 super(6) cells and 1 ml culture medium, respectively; 10-20% of the cells produced HCMV-specific antigens, while 6-13% produced infectious virus progeny. The number of HCMV-specific DNA copies ranged from 9 x 10 super(4) to 9 x 10 super(6) per 10 super(6) cells. Transmission electron microscopy confirmed the productive nature of HCMV infection. UKF-NB-4 super(AD169) cultures proliferated, with population doubling time ranging from 24.5 to 26.6 hr (19.5 to 20.3 hr for UKF-NB-4) and cell viability from 79% to 85% (91-96% for UKF-NB-4). Significantly lower amounts of tyrosine hydroxylase and decreased activity for dopamine- beta -hydroxylase than in uninfected cells were observed in UKF-NB-4 super(AD169) cells. However, the expression of N-myc oncoprotein was significantly increased in persistently infected cultures. Our results show that long-term productive HCMV infection of UKF-NB-4 cell line is associated with the modulation of phenotypic properties, which may be related to the biological behaviour of neuroblastoma cells.

Keywords: Cytomegalovirus ; Cytomegalovirus ; Neuroblastoma Cells ; Man ; Tumor Cells ; Phenotyping ; Neuroblastoma Cells ; Man ; Tumor Cells ; Phenotyping ; Neurovirology ; Virus Behavior in Cell Culture ; Tyrosine 3-Monooxygenase ; Dopamine Beta -Monooxygenase ; N-Myc Protein ; Tyrosine 3-Monooxygenase ; Dopamine Beta -Monooxygenase ; N-Myc Protein ; N-Myc Protein ; Dopamine Beta -Monooxygenase ; Tyrosine 3-Monooxygenase;

ISSN: 00207136

E-ISSN: 10970215

DOI: 10.1002/(SICI)1097-0215(19960103)65:1〈90::AID-IJC16〉3.0.CO;2-M

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7

Article

In vitro Antiviral Activity of Aphidicolin and its Derivates (2002)

Michaelis, Martin ; Langer, Klaus ; Vogel, Jens-Uwe ; [et al.]

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Language: English

In: Arzneimittelforschung, 5/2002, Vol.52(05), pp.393-399

ISSN: 0004-4172

E-ISSN: 1616-7066

DOI: 10.1055/s-0031-1299904

Source: Thieme Publishing Group (via CrossRef)

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In vitro antiviral activity of aphidicolin and its derivates. Synergistic effects of aphidicolin with other antiviral drugs (2002)

Michaelis, Martin ; Langer, Klaus ; Vogel, Jens-Uwe ; [et al.]

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Language: English

In: Arzneimittel-Forschung, 2002, Vol.52(5), pp.393-9

Description: Twenty derivatives of aphidicolin were tested against HSV (herpes simplex virus), HCMV (human cytomegalovirus) and adenovirus in vitro. In addition, the antiviral activity of aphidicolin (CAS 38966-21-1) in combination with aciclovir (CAS 59277-89-3) or cidofovir (CAS 113852-37-2) against HSV was determined. The antiviral effects were evaluated using plaque reduction assay in Vero cells or human Foreskin Fibroblasts (HFF) for HSV and HCMV, respectively. Combination indexes were calculated using the method of Chou and Talalay. Two derivatives (K14254 and K14266) that are considered to be prodrugs of aphidicolin were shown to inhibit HCMV and HSV replication comparably to aphidicolin. None of the tested substances inhibited adenovirus replication. Aphidicolin acted synergistically with aciclovir in a 1:1 molar ratio and with cidofovir in different ratios. Aphidicolin and its two antiviral active derivatives might represent useful additional tools for antiviral therapy of HSV and HCMV infections, especially in combination with clinically used drugs.

Keywords: Organophosphonates ; Antiviral Agents -- Pharmacology ; Aphidicolin -- Analogs & Derivatives

ISSN: 0004-4172

E-ISSN: 16167066

Source: MEDLINE/PubMed (U.S. National Library of Medicine)

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9

Article

Protein-free cell culture on polyvinyl formal surface (1992)

Cinatl, Jindrich ; Rabenau, Holger ; Cinatl, Jaroslav ; [et al.]

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Language: English

In: In Vitro Cellular & Developmental Biology - Animal, 1992, Vol.28(3), pp.147-148

Keywords: Biological sciences -- Biology -- Cytology ; Biological sciences -- Biology -- Cytology ; Biological sciences -- Biology -- Cytology ; Biological sciences -- Biology -- Cytology ; Applied sciences -- Laboratory techniques -- Culture techniques ; Physical sciences -- Chemistry -- Chemical compounds ; Physical sciences -- Chemistry -- Chemical compounds;

ISSN: 0883-8364

E-ISSN: 1543-706X

E-ISSN: 2327431X

DOI: 10.1007/BF02631082

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10

Article

A new assay system for quality control testing of a chemically defined medium (1989)

Cinatl, Jindrich ; Cinatl, Jaroslav ; Rabenau, Holger ; [et al.]

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Language: English

In: Journal of tissue culture methods, 1989, Vol.12(2), pp.67-72

Description: Mouse NCTC clone 929 L (L-929) cells were propagated continuously over 2 yr in protein-free Eagles's minimal essential medium (EMEM). The cells designated L-929-WS were used for quality-control testing of protein-free EMEM as a model for more general medium quality tests. The parameter for the suggested quality control assay was the growth-promoting activity of the medium for L-929-WS cells. As an example of quality tests we assayed the growth promoting activity of EMEM exposed to various conditions of storage time and temperature. We established the sensitivity of the new assay by comparing it to the original cells L-929 grown in EMEM supplemented with 10% fetal bovine serum (FBS). The assay system using L-929-WS cells propagated continuously in protein-free medium proved to be much more sensitive. The sensitivity, however, was abolished by the addition of FBS in a concentration as low as 1% to a culture medium.

Keywords: protein-free medium ; quality control testing ; growth-promoting activity

ISSN: 0271-8057

E-ISSN: 1573-0603

DOI: 10.1007/BF01404646

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Kooperativer Bibliotheksverbund

Berlin Brandenburg