G. Pauli - Academia.edu (original) (raw)
Papers by G. Pauli
Berliner und Münchener tierärztliche Wochenschrift, 1984
Mund-, Kiefer- und Gesichtschirurgie, 2003
ABSTRACT HintergrundDie humane Zytomegalievirusinfektion (HCMV-Infektion) kann bei immunkompromit... more ABSTRACT HintergrundDie humane Zytomegalievirusinfektion (HCMV-Infektion) kann bei immunkompromittierten Patienten, so auch nach Knochenmark- (KMT) und Stammzelltransplantation (SCT), Ursache für schwere, zum Teil lebensbedrohliche Erkrankungen sein. Das Risiko einer HCMV-Erkrankung wird vor der Transplantation durch die Bestimmung von HCMV-Antikörpern im Blut von Empfänger (E) und Spender (S) ermittelt. Virusträger sind Anti-HCMV-IgG-Antikörper-positiv. Es ergeben sich die folgenden Viruskonstellationen: Gruppe 1 (E+/S+), Gruppe 2 (E+/S−), Gruppe 3 (E−/S+) und Gruppe 4 (E−/S−). ZielZiel der Untersuchung war die qualitative und quantitative Bestimmung der HCMV-Viruslast im Speichel von Patienten mit KMT und SCT. Patienten und MethodeZur Bestimmung des HCMV wurde eine TaqMan®-PCR etabliert. Ruhespeichel von 20 Patienten wurde vor der KMT und SCT, während der Konditionierungsphase sowie nach der Transplantation entnommen. Die DNA wurde isoliert und mit der TaqMan®-PCR auf HCMV-DNA untersucht. ErgebnisseHCMV-DNA konnte bei insgesamt 7 Patienten isoliert werden. Bei allen 4 Patienten der Gruppe 1 (E+/S+) wurde HCMV-DNA nachgewiesen. Im Speichel der 7 Patienten der Gruppe 2 (E+/S−) ließ sich nur in 3 Fällen HCMV-DNA bestimmen. Bei dem einen Patienten der Gruppe 3 (E−/S+) und den acht Patienten der Gruppe 4 (E−/S−) wurde keine HCMV-DNA isoliert. SchlussfolgerungDie TaqMan®-PCR erbringt einen sicheren quantitativen Nachweis von HCMV-DNA im Speichel. Bei 3 Patienten, die Anti-HCMV-IgG-positiv waren und ein Anti-CMV-IgG-negatives Transplantat erhalten haben, konnte HCMV-DNA bestimmt werden, wohingegen in der Gruppe HCMV-negativer Spender und HCMV-negativer Empfänger das Virus bei keinem der Patienten nachweisbar war. Das Risiko der HCMV-Reaktivierung ist somit größer als das einer Neuinfektion. BackgroundHuman cytomegalovirus (HCMV) infection is associated with severe and life-threatening diseases in immunocompromised patients, especially after bone marrow (BM) and stem cell (SC) transplantation. Prior to transplantation the potential risk of HCMV disease is therefore determined by HCMV-antibody blood testing of transplant donor (D) and recipient (R). Virus carriers are positive for anti-CMV-IgG. Virus patterns are distinguished as follows: group 1 (D+/R+), group 2 (D−/R+), group 3 (D+/R−), and group 4 (D−/R−). AimThe aim of this study was qualitative and quantitative determination of the HCMV DNA load in saliva of BM and SC transplantation patients. Patients and methodUnstimulated saliva was collected from 20 patients prior to BM and SC transplantation, during the time of conditioning, and after transplantation. DNA was isolated and analyzed for evidence of HCMV DNA with TaqMan PCR. ResultsHCMV DNA was isolated in seven cases. In all group 1 patients (D+/R+) HCMV DNA could be demonstrated. Only three of seven group 2 patients (D−/R+) were positive for HCMV DNA. The only group 3 patient (D+/R−) and all eight group 4 patients (D−/R−) were negative. ConclusionTaqMan PCR is a reliable method for HCMV DNA quantification. In three patients (anti-HCMV-IgG positive) who received an anti-CMV-IgG negative transplant HCMV DNA was isolated. In contrast, no HCMV-DNA was evident in HCMV-negative patients who received an HCMV-negative transplant. Accordingly, the risk of HCMV reactivation is more probable than the risk of reinfection. SchlüsselwörterHumanes Zytomegalievirus–TaqMan®-PCR–Knochenmark- und Stammzelltransplantation KeywordsHuman cytomegalovirus–TaqMan PCR–Bone marrow and stem cell transplantation
Virology, 1975
The properties of the purified DNA-polymerase of these viruses were analyzed with the following s... more The properties of the purified DNA-polymerase of these viruses were analyzed with the following significant findings: 1) Unfrozen, purified disrupted REV, but not purified REV-DNA-polymerase is enzymatically active using natural viral RNA as template-primer; 2) the ...
Veterinary Pathology, 2006
An epizootic infection was observed in a colony of 80 New World monkeys consisting of various spe... more An epizootic infection was observed in a colony of 80 New World monkeys consisting of various species including a group of marmosets and Saguinus species. During the summer and autumn of 2002, 30 animals died of unknown diseases. Six animals were sent to the German Primate Center for investigation of the cause of death. A complete pathologic and histologic investigation was carried out. The animals exhibited erosive-ulcerative lesions of the oral mucous membranes. Advanced stages of the disease were characterised by hemorrhagic lesions on the skin distributed randomly over the body, but principally on the face, scrotal region, soles, and palms. Electron microscopy revealed virus particles with orthopox-like morphology within intracytoplasmic inclusions in epithelial cells. The DNA samples from various tissues were analyzed by use of a set of orthopox virus-specific, real-time polymerase chain reaction assays. Amplification products were sequenced to define the virus more precisely. ...
Transfusion Medicine and Hemotherapy, 1999
Summary Background: In order to evaluate the prevalence of human T-cell lymphotropic virus type I... more Summary Background: In order to evaluate the prevalence of human T-cell lymphotropic virus type I and II (HTLV-I/-II) infections in Germany, the Bavarian blood donor population was screened. Material and Methods:Between 1991 and 1994 around 1.5 million donations from about 375,000 blood donors of the Bavarian Red Cross (BRK) were tested for HTLV-I/-II in an antibody screening test. Sera repeatedly
Journal of General Virology, 1992
Biologicals, 1996
No abstract
Biological Conservation, 2006
Until recently, the focus of great ape behavioural and ecological research has been distinct from... more Until recently, the focus of great ape behavioural and ecological research has been distinct from the focus of scientists working in medical and veterinary sciences. More scientists are calling for a connection between medical and field research due to recent disease ...
Archives of Virology, 1978
Gp85 the major envelope glycoprotein of Rous sarcoma virus can be released from intact by virus t... more Gp85 the major envelope glycoprotein of Rous sarcoma virus can be released from intact by virus treatment with 2-mercaptoethanol. Attempts were made to rcassociate released gp85 to 2-mercaptoethanol treated virus. In order to study the biological activity and to analyse the reaction product with biochemical methods gp85 from a subgroup different from the recipient virus was used. Successful in vitro phenotypic mixing could be showm. The biochemical analysis, however, revealed that no reassociation of gp85 and gp35 could be obtained. The biological activity was due to a strong association of whole gp 85-gp 35 molecules derived from the gp85 preparation to the recipient virus. These molecules were present as a minor contamination in the gp 85 preparation.
Archives of Virology, 1989
Monoclonal antibodies directed against a surface glycoprotein of the bovine herpes virus type 2 (... more Monoclonal antibodies directed against a surface glycoprotein of the bovine herpes virus type 2 (BHV-2, bovine herpes mammillitis virus) recognize also determinants of the major glycoprotein gB of the human herpes simplex virus type 1 (HSV-1). Cross-reacting antigens of the virions and in infected cells were localized with immunocytochemical methods, immunofluorescence as well as pre-embedding and cryoultramicrotomy immune electron microscopy. All antibodies stain to different degrees cell free BHV-2 and HSV-1 virions. In the cell two predominant staining patterns could be observed indicating that expression of epitopes is dependent upon the cell compartment: (i) staining of cytoplasmic membranes and enveloped particles within membrane systems and (ii) staining of intranuclear antigens. Antibodies tagging intranuclear antigens react with moderately dense material or with the periphery of nucleocapsids. This unexpected result is interpreted in terms of two hypotheses: (1) presence of common epitopes on two entirely different herpesvirus proteins conserved in HSV-1 and BHV-2 and (2) transport of gB or its precursor into the nucleus.
Applied and Environmental Microbiology, 2008
After terrorists attacked the United States in 2001, the appearance of letters and other objects ... more After terrorists attacked the United States in 2001, the appearance of letters and other objects containing powdery substances with unknown potentials for biological threat focused attention on the speed, sensitivity, and reliability of diagnostic methods. This study summarizes the abilities and limitations of real-time PCR, electron microscopy (EM), and virus isolation when used to detect potential bioweapons. In particular, we investigated the inhibitory influences of different common household products present in environmental specimens on PCR yield, EM detection, and virus isolation. We used vaccinia virus as a model for orthopoxviruses by spiking it into specimens. In the second part of the study, we describe modifications of diagnostic methods to overcome inhibitory effects. A variety of PCR amplification enhancers, DNA extraction protocols, and applications of internal controls were evaluated to improve diagnostic simplicity, speed, and reliability. As a result, we strongly recommend using at least two different frontline techniques in parallel, e.g., EM and PCR. A positive result obtained by any one of these techniques should be followed by a biological method to confirm the putative diagnosis. Confirmatory methods include virus isolation followed by an agent-specific immunofluorescence assay to confirm the presence of replication-competent particles.
Leukemia, 1999
Figure 1 HLA A-B-C antigen expression during in vitro differentiation with ATRA 10 −7 m. APL cell... more Figure 1 HLA A-B-C antigen expression during in vitro differentiation with ATRA 10 −7 m. APL cells isolated from patients harboring either bcr1 or bcr3 PMLRAR␣ subtype were cultured 6 days in the absence (grey bars) or in the presence of ATRA 10 −7 m (white bars). Each bar represents the mean ± s.d. of HLA A-B-C expression for all patients (n = 10). First non-imported HTLV-1 positive adult T cell leukemia/lymphoma (ATLL) in Germany TO THE EDITOR We report the first non-imported HTLV-1 positive adult T cell leukemia/lymphoma in Germany. A 41-year-old German mother of a healthy 12-year-old girl was admitted for generalized lymphadenopathy, splenomegaly, a macular exanthema and fever, all having developed within the past 2 weeks. The patient was confused and somnolent. No other abnormalities were observed at physical examination. Radiological assessment revealed a bilateral mediastinal lymph node enlargement. A magnetic resonance imaging (MRI) of the skull disclosed no abnormalities. Laboratory analyses displayed a normal hemoglobin and thrombocyte count, leukocytes were 45 G/l with 36% atypical flower-like lymphoid cells, creatinine was 2.04 (normal range: 0.3-1.0) mg/dl and calcium was 4.43 (2.10-2.60) mmol/l, lactate dehydrogenase was also elevated with 800 (230-400) U/l. Immunophenotyping of peripheral blood mononuclear cells identified the atypical cells as CD3 + , CD4 low+ , CD25 + , CD38 + , CD7 − , CD8 − mature T cells. Histologic examination of a lymph node biopsy showed an architecture destroyed by a CD3 + and BCL-2 + , CD20 − , kappa − , lambda − , CD21, CD30 − and CD15 − peripheral T cell lymphoma with a growth rate of 40%. The bone marrow biopsy disclosed an extended infiltration by the malignant T cell population. Pleocytosis and blasts were also seen in the cerebrospinal fluid (CSF). Antibodies against HTLV-1/2 were found in CSF and serum. A Western blot identified the characteristic anti-HTLV-1 band pattern both in patient's serum and CSF. Type-specific PCR analysis targeted to fragments of pol identified HTLV-1 as the integrated virion. High serum levels of soluble interleukin-2 receptor (s-IL-2R) as well as soluble tumor necrosis factor receptor type II (s-TNF R II) were recorded (Immulite, DPC Biermann, Germany). The patient had no history of i.v. drug abuse, blood transfusion or contact to residents of HTLV-1 endemic areas. The serology for HIV-1/2 and HBV was negative, while IgG antibodies were detected against VZV, CMV and EBV. Cytogenetic studies of peripheral blood lymphocytes (PBL) revealed consistent structural
Biologicals, 1997
The antiviral activity of surfactin, a cyclic lipopeptide antibiotic and biosurfactant produced b... more The antiviral activity of surfactin, a cyclic lipopeptide antibiotic and biosurfactant produced by Bacillus subtilis, was determined for a broad spectrum of different viruses, Semliki Forest virus (SFV), herpes simplex virus (HSV-1, HSV-2), suid herpes virus (SHV-1), vesicular stomatitis virus (VSV), simian immunodeficiency virus (SIV), feline calicivirus (FCV), murine encephalomyocarditis virus (EMCV). In vitro experiments showed biphasic virus inactivation kinetics for enveloped viruses during treatment. Inactivation of enveloped viruses, especially herpes- and retroviruses, was much more efficient than that of non-enveloped viruses. For those viruses susceptible to its action, surfactin was active at 25 microM in medium containing 5% fetal calf serum (FCS). Concentrations up to 80 microM of surfactin led to a titre reduction of >4.4 log10 CCID50/ml for HSV-1 in 15 min and for SIV and VSV in 60 min. The inactivation rate increased linearly with the incubation temperature by a factor 2.4/10 degrees C and logarithmically with the concentration. Serum components, probably proteins and/or lipids, influence the effective surfactin concentration. A disruption of the viral lipid membrane and partially of the capsid was observed by electron microscopy. These findings suggest that the antiviral action, postulated also in other investigations, seems to be due to a physicochemical interaction of the membrane-active surfactant with the virus lipid membrane. Surfactin may be useful for application in virus safety enhancement of biotechnological and pharmaceutical products.
Bundesgesundheitsblatt, 1997
Journal of Virological Methods, 1997
Eleven German laboratories and one Swiss laboratory initiated a quality assessment study to evalu... more Eleven German laboratories and one Swiss laboratory initiated a quality assessment study to evaluate the specificity and sensitivity of their polymerase chain reaction (PCR) for detection of HIV-1 DNA. Following its own PCR protocols, each laboratory tested a panel of ten coded samples consisting of cell pellets containing 0, 0.1, 1, 10, 10(2), 10(3) and 10(4) ACH-2 cells per 1.5 x 10(5) uninfected peripheral blood mononuclear cells. Of the twelve participating laboratories, three reported correct results for the dilution series as well as for uninfected specimens. One or more classification errors were recorded for 12% of the samples for which the diagnosis was expected to be positive or negative. Samples containing 10 copies of the target template were correctly reported by eleven of the twelve participants. The average sensitivity was 97%. The results of the study revealed no significant differences between the Amplicor kit and in-house procedures. Most of the classification errors occurred in specimens from HIV-negative samples. Out of 36 negative samples, 5 were reported false positive, showing that contamination remains a problem for some laboratories, regardless of the PCR test performed. Careful laboratory techniques and internal as well as external quality control procedures will help avoiding carryover contamination.
Archives of Virology, 1992
The expression of the capsid antigen (CA) and the two regulatory proteins nef and vpu as well as ... more The expression of the capsid antigen (CA) and the two regulatory proteins nef and vpu as well as the CD4 cell surface receptor was followed in HIV-infected lymphoid and promonocytic cells. In the lytic phase of infection all three viral proteins were expressed; production of these proteins coincided with the increase of CA antigen and infectious virus in culture supernatants and with prominent cytopathic effects. After selection of persistently infected cells, the number of lymphoid cells expressing detectable levels of nefdecreased to zero; the number of cells positive for CA ranged between 40 to 70%. In chronically infected promonocytic cells nef and vpu expression was reduced to undetectable levels, whereas most of the cells accumulated CA intracellularly. Infectious cell free virus and CA in the supernatant of promonocytic cells had low titers. CD4 surface expression declined in all cell lines investigated before cell free virus was detectable. down regulate CD4 expression on the surface of T-cells [19], but not CD4 162 T. Schneider et al. protein synthesis [17]. On the other side positive effects of nef on virus replication have been reported . Investigations on the function of vpu suggest that this HIV-1 specific protein is involved in final steps of virus assembly and release from cells .
Archives of Virology, 1987
A panel of monoclonal antibodies to glyeoprotein B (gB) of herpes simplex virus 1 (HSV-1) reacted... more A panel of monoclonal antibodies to glyeoprotein B (gB) of herpes simplex virus 1 (HSV-1) reacted in immune precipitation and in immunofluorescence tests with a glyeoprotein 130,000 in apparent molecular weight specified by bovine herpesvirus 2 (BHV-2). Two cross reactive antibodies neutralized the heterologous virus. These results suggest that the BHV-2 glyeoprotein is the homologue of HSV-1 gB and shares structural and functional domains expressed on the gB gene product.
Berliner und Münchener tierärztliche Wochenschrift, 1984
Mund-, Kiefer- und Gesichtschirurgie, 2003
ABSTRACT HintergrundDie humane Zytomegalievirusinfektion (HCMV-Infektion) kann bei immunkompromit... more ABSTRACT HintergrundDie humane Zytomegalievirusinfektion (HCMV-Infektion) kann bei immunkompromittierten Patienten, so auch nach Knochenmark- (KMT) und Stammzelltransplantation (SCT), Ursache für schwere, zum Teil lebensbedrohliche Erkrankungen sein. Das Risiko einer HCMV-Erkrankung wird vor der Transplantation durch die Bestimmung von HCMV-Antikörpern im Blut von Empfänger (E) und Spender (S) ermittelt. Virusträger sind Anti-HCMV-IgG-Antikörper-positiv. Es ergeben sich die folgenden Viruskonstellationen: Gruppe 1 (E+/S+), Gruppe 2 (E+/S−), Gruppe 3 (E−/S+) und Gruppe 4 (E−/S−). ZielZiel der Untersuchung war die qualitative und quantitative Bestimmung der HCMV-Viruslast im Speichel von Patienten mit KMT und SCT. Patienten und MethodeZur Bestimmung des HCMV wurde eine TaqMan®-PCR etabliert. Ruhespeichel von 20 Patienten wurde vor der KMT und SCT, während der Konditionierungsphase sowie nach der Transplantation entnommen. Die DNA wurde isoliert und mit der TaqMan®-PCR auf HCMV-DNA untersucht. ErgebnisseHCMV-DNA konnte bei insgesamt 7 Patienten isoliert werden. Bei allen 4 Patienten der Gruppe 1 (E+/S+) wurde HCMV-DNA nachgewiesen. Im Speichel der 7 Patienten der Gruppe 2 (E+/S−) ließ sich nur in 3 Fällen HCMV-DNA bestimmen. Bei dem einen Patienten der Gruppe 3 (E−/S+) und den acht Patienten der Gruppe 4 (E−/S−) wurde keine HCMV-DNA isoliert. SchlussfolgerungDie TaqMan®-PCR erbringt einen sicheren quantitativen Nachweis von HCMV-DNA im Speichel. Bei 3 Patienten, die Anti-HCMV-IgG-positiv waren und ein Anti-CMV-IgG-negatives Transplantat erhalten haben, konnte HCMV-DNA bestimmt werden, wohingegen in der Gruppe HCMV-negativer Spender und HCMV-negativer Empfänger das Virus bei keinem der Patienten nachweisbar war. Das Risiko der HCMV-Reaktivierung ist somit größer als das einer Neuinfektion. BackgroundHuman cytomegalovirus (HCMV) infection is associated with severe and life-threatening diseases in immunocompromised patients, especially after bone marrow (BM) and stem cell (SC) transplantation. Prior to transplantation the potential risk of HCMV disease is therefore determined by HCMV-antibody blood testing of transplant donor (D) and recipient (R). Virus carriers are positive for anti-CMV-IgG. Virus patterns are distinguished as follows: group 1 (D+/R+), group 2 (D−/R+), group 3 (D+/R−), and group 4 (D−/R−). AimThe aim of this study was qualitative and quantitative determination of the HCMV DNA load in saliva of BM and SC transplantation patients. Patients and methodUnstimulated saliva was collected from 20 patients prior to BM and SC transplantation, during the time of conditioning, and after transplantation. DNA was isolated and analyzed for evidence of HCMV DNA with TaqMan PCR. ResultsHCMV DNA was isolated in seven cases. In all group 1 patients (D+/R+) HCMV DNA could be demonstrated. Only three of seven group 2 patients (D−/R+) were positive for HCMV DNA. The only group 3 patient (D+/R−) and all eight group 4 patients (D−/R−) were negative. ConclusionTaqMan PCR is a reliable method for HCMV DNA quantification. In three patients (anti-HCMV-IgG positive) who received an anti-CMV-IgG negative transplant HCMV DNA was isolated. In contrast, no HCMV-DNA was evident in HCMV-negative patients who received an HCMV-negative transplant. Accordingly, the risk of HCMV reactivation is more probable than the risk of reinfection. SchlüsselwörterHumanes Zytomegalievirus–TaqMan®-PCR–Knochenmark- und Stammzelltransplantation KeywordsHuman cytomegalovirus–TaqMan PCR–Bone marrow and stem cell transplantation
Virology, 1975
The properties of the purified DNA-polymerase of these viruses were analyzed with the following s... more The properties of the purified DNA-polymerase of these viruses were analyzed with the following significant findings: 1) Unfrozen, purified disrupted REV, but not purified REV-DNA-polymerase is enzymatically active using natural viral RNA as template-primer; 2) the ...
Veterinary Pathology, 2006
An epizootic infection was observed in a colony of 80 New World monkeys consisting of various spe... more An epizootic infection was observed in a colony of 80 New World monkeys consisting of various species including a group of marmosets and Saguinus species. During the summer and autumn of 2002, 30 animals died of unknown diseases. Six animals were sent to the German Primate Center for investigation of the cause of death. A complete pathologic and histologic investigation was carried out. The animals exhibited erosive-ulcerative lesions of the oral mucous membranes. Advanced stages of the disease were characterised by hemorrhagic lesions on the skin distributed randomly over the body, but principally on the face, scrotal region, soles, and palms. Electron microscopy revealed virus particles with orthopox-like morphology within intracytoplasmic inclusions in epithelial cells. The DNA samples from various tissues were analyzed by use of a set of orthopox virus-specific, real-time polymerase chain reaction assays. Amplification products were sequenced to define the virus more precisely. ...
Transfusion Medicine and Hemotherapy, 1999
Summary Background: In order to evaluate the prevalence of human T-cell lymphotropic virus type I... more Summary Background: In order to evaluate the prevalence of human T-cell lymphotropic virus type I and II (HTLV-I/-II) infections in Germany, the Bavarian blood donor population was screened. Material and Methods:Between 1991 and 1994 around 1.5 million donations from about 375,000 blood donors of the Bavarian Red Cross (BRK) were tested for HTLV-I/-II in an antibody screening test. Sera repeatedly
Journal of General Virology, 1992
Biologicals, 1996
No abstract
Biological Conservation, 2006
Until recently, the focus of great ape behavioural and ecological research has been distinct from... more Until recently, the focus of great ape behavioural and ecological research has been distinct from the focus of scientists working in medical and veterinary sciences. More scientists are calling for a connection between medical and field research due to recent disease ...
Archives of Virology, 1978
Gp85 the major envelope glycoprotein of Rous sarcoma virus can be released from intact by virus t... more Gp85 the major envelope glycoprotein of Rous sarcoma virus can be released from intact by virus treatment with 2-mercaptoethanol. Attempts were made to rcassociate released gp85 to 2-mercaptoethanol treated virus. In order to study the biological activity and to analyse the reaction product with biochemical methods gp85 from a subgroup different from the recipient virus was used. Successful in vitro phenotypic mixing could be showm. The biochemical analysis, however, revealed that no reassociation of gp85 and gp35 could be obtained. The biological activity was due to a strong association of whole gp 85-gp 35 molecules derived from the gp85 preparation to the recipient virus. These molecules were present as a minor contamination in the gp 85 preparation.
Archives of Virology, 1989
Monoclonal antibodies directed against a surface glycoprotein of the bovine herpes virus type 2 (... more Monoclonal antibodies directed against a surface glycoprotein of the bovine herpes virus type 2 (BHV-2, bovine herpes mammillitis virus) recognize also determinants of the major glycoprotein gB of the human herpes simplex virus type 1 (HSV-1). Cross-reacting antigens of the virions and in infected cells were localized with immunocytochemical methods, immunofluorescence as well as pre-embedding and cryoultramicrotomy immune electron microscopy. All antibodies stain to different degrees cell free BHV-2 and HSV-1 virions. In the cell two predominant staining patterns could be observed indicating that expression of epitopes is dependent upon the cell compartment: (i) staining of cytoplasmic membranes and enveloped particles within membrane systems and (ii) staining of intranuclear antigens. Antibodies tagging intranuclear antigens react with moderately dense material or with the periphery of nucleocapsids. This unexpected result is interpreted in terms of two hypotheses: (1) presence of common epitopes on two entirely different herpesvirus proteins conserved in HSV-1 and BHV-2 and (2) transport of gB or its precursor into the nucleus.
Applied and Environmental Microbiology, 2008
After terrorists attacked the United States in 2001, the appearance of letters and other objects ... more After terrorists attacked the United States in 2001, the appearance of letters and other objects containing powdery substances with unknown potentials for biological threat focused attention on the speed, sensitivity, and reliability of diagnostic methods. This study summarizes the abilities and limitations of real-time PCR, electron microscopy (EM), and virus isolation when used to detect potential bioweapons. In particular, we investigated the inhibitory influences of different common household products present in environmental specimens on PCR yield, EM detection, and virus isolation. We used vaccinia virus as a model for orthopoxviruses by spiking it into specimens. In the second part of the study, we describe modifications of diagnostic methods to overcome inhibitory effects. A variety of PCR amplification enhancers, DNA extraction protocols, and applications of internal controls were evaluated to improve diagnostic simplicity, speed, and reliability. As a result, we strongly recommend using at least two different frontline techniques in parallel, e.g., EM and PCR. A positive result obtained by any one of these techniques should be followed by a biological method to confirm the putative diagnosis. Confirmatory methods include virus isolation followed by an agent-specific immunofluorescence assay to confirm the presence of replication-competent particles.
Leukemia, 1999
Figure 1 HLA A-B-C antigen expression during in vitro differentiation with ATRA 10 −7 m. APL cell... more Figure 1 HLA A-B-C antigen expression during in vitro differentiation with ATRA 10 −7 m. APL cells isolated from patients harboring either bcr1 or bcr3 PMLRAR␣ subtype were cultured 6 days in the absence (grey bars) or in the presence of ATRA 10 −7 m (white bars). Each bar represents the mean ± s.d. of HLA A-B-C expression for all patients (n = 10). First non-imported HTLV-1 positive adult T cell leukemia/lymphoma (ATLL) in Germany TO THE EDITOR We report the first non-imported HTLV-1 positive adult T cell leukemia/lymphoma in Germany. A 41-year-old German mother of a healthy 12-year-old girl was admitted for generalized lymphadenopathy, splenomegaly, a macular exanthema and fever, all having developed within the past 2 weeks. The patient was confused and somnolent. No other abnormalities were observed at physical examination. Radiological assessment revealed a bilateral mediastinal lymph node enlargement. A magnetic resonance imaging (MRI) of the skull disclosed no abnormalities. Laboratory analyses displayed a normal hemoglobin and thrombocyte count, leukocytes were 45 G/l with 36% atypical flower-like lymphoid cells, creatinine was 2.04 (normal range: 0.3-1.0) mg/dl and calcium was 4.43 (2.10-2.60) mmol/l, lactate dehydrogenase was also elevated with 800 (230-400) U/l. Immunophenotyping of peripheral blood mononuclear cells identified the atypical cells as CD3 + , CD4 low+ , CD25 + , CD38 + , CD7 − , CD8 − mature T cells. Histologic examination of a lymph node biopsy showed an architecture destroyed by a CD3 + and BCL-2 + , CD20 − , kappa − , lambda − , CD21, CD30 − and CD15 − peripheral T cell lymphoma with a growth rate of 40%. The bone marrow biopsy disclosed an extended infiltration by the malignant T cell population. Pleocytosis and blasts were also seen in the cerebrospinal fluid (CSF). Antibodies against HTLV-1/2 were found in CSF and serum. A Western blot identified the characteristic anti-HTLV-1 band pattern both in patient's serum and CSF. Type-specific PCR analysis targeted to fragments of pol identified HTLV-1 as the integrated virion. High serum levels of soluble interleukin-2 receptor (s-IL-2R) as well as soluble tumor necrosis factor receptor type II (s-TNF R II) were recorded (Immulite, DPC Biermann, Germany). The patient had no history of i.v. drug abuse, blood transfusion or contact to residents of HTLV-1 endemic areas. The serology for HIV-1/2 and HBV was negative, while IgG antibodies were detected against VZV, CMV and EBV. Cytogenetic studies of peripheral blood lymphocytes (PBL) revealed consistent structural
Biologicals, 1997
The antiviral activity of surfactin, a cyclic lipopeptide antibiotic and biosurfactant produced b... more The antiviral activity of surfactin, a cyclic lipopeptide antibiotic and biosurfactant produced by Bacillus subtilis, was determined for a broad spectrum of different viruses, Semliki Forest virus (SFV), herpes simplex virus (HSV-1, HSV-2), suid herpes virus (SHV-1), vesicular stomatitis virus (VSV), simian immunodeficiency virus (SIV), feline calicivirus (FCV), murine encephalomyocarditis virus (EMCV). In vitro experiments showed biphasic virus inactivation kinetics for enveloped viruses during treatment. Inactivation of enveloped viruses, especially herpes- and retroviruses, was much more efficient than that of non-enveloped viruses. For those viruses susceptible to its action, surfactin was active at 25 microM in medium containing 5% fetal calf serum (FCS). Concentrations up to 80 microM of surfactin led to a titre reduction of >4.4 log10 CCID50/ml for HSV-1 in 15 min and for SIV and VSV in 60 min. The inactivation rate increased linearly with the incubation temperature by a factor 2.4/10 degrees C and logarithmically with the concentration. Serum components, probably proteins and/or lipids, influence the effective surfactin concentration. A disruption of the viral lipid membrane and partially of the capsid was observed by electron microscopy. These findings suggest that the antiviral action, postulated also in other investigations, seems to be due to a physicochemical interaction of the membrane-active surfactant with the virus lipid membrane. Surfactin may be useful for application in virus safety enhancement of biotechnological and pharmaceutical products.
Bundesgesundheitsblatt, 1997
Journal of Virological Methods, 1997
Eleven German laboratories and one Swiss laboratory initiated a quality assessment study to evalu... more Eleven German laboratories and one Swiss laboratory initiated a quality assessment study to evaluate the specificity and sensitivity of their polymerase chain reaction (PCR) for detection of HIV-1 DNA. Following its own PCR protocols, each laboratory tested a panel of ten coded samples consisting of cell pellets containing 0, 0.1, 1, 10, 10(2), 10(3) and 10(4) ACH-2 cells per 1.5 x 10(5) uninfected peripheral blood mononuclear cells. Of the twelve participating laboratories, three reported correct results for the dilution series as well as for uninfected specimens. One or more classification errors were recorded for 12% of the samples for which the diagnosis was expected to be positive or negative. Samples containing 10 copies of the target template were correctly reported by eleven of the twelve participants. The average sensitivity was 97%. The results of the study revealed no significant differences between the Amplicor kit and in-house procedures. Most of the classification errors occurred in specimens from HIV-negative samples. Out of 36 negative samples, 5 were reported false positive, showing that contamination remains a problem for some laboratories, regardless of the PCR test performed. Careful laboratory techniques and internal as well as external quality control procedures will help avoiding carryover contamination.
Archives of Virology, 1992
The expression of the capsid antigen (CA) and the two regulatory proteins nef and vpu as well as ... more The expression of the capsid antigen (CA) and the two regulatory proteins nef and vpu as well as the CD4 cell surface receptor was followed in HIV-infected lymphoid and promonocytic cells. In the lytic phase of infection all three viral proteins were expressed; production of these proteins coincided with the increase of CA antigen and infectious virus in culture supernatants and with prominent cytopathic effects. After selection of persistently infected cells, the number of lymphoid cells expressing detectable levels of nefdecreased to zero; the number of cells positive for CA ranged between 40 to 70%. In chronically infected promonocytic cells nef and vpu expression was reduced to undetectable levels, whereas most of the cells accumulated CA intracellularly. Infectious cell free virus and CA in the supernatant of promonocytic cells had low titers. CD4 surface expression declined in all cell lines investigated before cell free virus was detectable. down regulate CD4 expression on the surface of T-cells [19], but not CD4 162 T. Schneider et al. protein synthesis [17]. On the other side positive effects of nef on virus replication have been reported . Investigations on the function of vpu suggest that this HIV-1 specific protein is involved in final steps of virus assembly and release from cells .
Archives of Virology, 1987
A panel of monoclonal antibodies to glyeoprotein B (gB) of herpes simplex virus 1 (HSV-1) reacted... more A panel of monoclonal antibodies to glyeoprotein B (gB) of herpes simplex virus 1 (HSV-1) reacted in immune precipitation and in immunofluorescence tests with a glyeoprotein 130,000 in apparent molecular weight specified by bovine herpesvirus 2 (BHV-2). Two cross reactive antibodies neutralized the heterologous virus. These results suggest that the BHV-2 glyeoprotein is the homologue of HSV-1 gB and shares structural and functional domains expressed on the gB gene product.