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Biomedical subjects

H Wolf

Publications and source records attributed to H Wolf.

At least 271 records · Page 15Linked to original sources
PubMed1995

Antibodies to parvovirus B19 NS-1 protein in infected individuals.

Human parvovirus B19 is the aetiological agent of the common childhood disease erythema infectiosum (fifth disease). The infection is usually benign and self-limiting, but in adults cases of severe arthritis which may persist for years have been reported. Neutralizing antibodies directed against the structural proteins are usually produced shortly after the infection. The immune response against the third major protein, the non-structural protein NS-1, of parvovirus B19 has not been characterized so far. We cloned and expressed the full-length NS-1 protein and fragments thereof in Escherichia coli. The purified recombinant proteins were used to investigate the presence of antibodies to the NS-1 protein in sera from patients with parvovirus B19 infection. Specific antibodies could be detected in sera from patients suffering from severe parvovirus B19-associated arthritis using Western blot analysis and an ELISA. Sera from patients with acute or past infection without complications did not contain detectable levels of immunoglobulin to NS-1. The use of subfragments of the NS-1 protein allowed localization of the antigenic domains in the carboxy-terminal region of the protein.

Antibodies, Viral↗
PubMed1995

Epstein-Barr virus lytic replication is controlled by posttranscriptional negative regulation of BZLF1.

Regulation of the immediate-early gene BZLF1 is assumed to play a key role in triggering the lytic replication of Epstein-Barr virus (EBV). The expression of BZLF1 is regulated on multiple levels, including control of transcription by several positive and negative cis-acting elements as well as posttranslational modifications and protein-protein interactions. Localization of BZLF1 on one strand of the genome and the latent EBNA1 transcription unit on the complementary strand suggests a regulatory mechanism via hybridization of antisense RNA. With a plasmid encoding a defective BZLF1 RNA, which could not be translated, we were able to induce expression of endogenous BZLF1 gene product Zta and other proteins of the lytic cycle. Our data show for the first time that latent replication is stabilized by negative regulation of an immediate-early gene of the lytic cycle by a posttranscriptional mechanism. This might be a common theme of herpes simplex virus and EBV latency.

Base Sequence↗
PubMed1995

Specific cytotoxic T lymphocytes recognize the immediate-early transactivator Zta of Epstein-Barr virus.

We identified the immediate-early transactivator Zta of Epstein-Barr virus as a target for specific cytotoxic T lymphocytes (CTL). Cells pulsed with overlapping synthetic peptides representing the entire amino acid sequence of Zta proved to be efficient for the in vitro stimulation of Zta-specific CTL in several donors. With peptide-pulsed target cells, we found that CTL from several donors recognize a peptide comprising 15 amino acids. The immune response against this peptide exerted by CTL lines from different donors was found to be restricted by two different molecules of the major histocompatibility complex: HLA-B8 and HLA-Cw6. The latter molecule could for the first time be identified as a restricting element for a CTL response. The epitope of the HLA-B8-restricted CTL could be mapped to an octameric sequence between amino acid positions 190 and 197 of the Zta protein, whereas the minimal epitope of HLA-Cw6-restricted CTL consists of 11 to 15 residues between positions 187 and 201. Thus, the HLA-B8 and HLA-Cw6 epitopes widely overlap but are not completely identical. In vitro stimulation of blood lymphocytes from a panel of HLA-B8-positive or HLA-Cw6-positive virus carriers, using autologous cells pulsed with the Zta peptides comprising the HLA-B8 or HLA-Cw6 epitope, respectively, revealed in both cases that most of these donors developed a Zta-specific cytotoxic activity. These data, as well as the high spread of the major histocompatibility complex molecules HLA-B8 and HLA-Cw6 in most populations, suggest that an efficient CTL response directed against gene products of the immediate-early group of the lytic cycle exists in vivo in a considerable portion of virus carriers. A CTL response against proteins expressed immediately after the switch into the lytic cycle could eliminate lytically activated cells at an early stage and would thus efficiently prevent the production and release of progeny virions.

Amino Acid Sequence↗
PubMed1995

Nonspiking local interneurons in insect leg motor control. I. Common layout and species-specific response properties of femur-tibia joint control pathways in stick insect and locust.

1. Locusts (Locusta migratoria) and stick insects (Carausius morosus) exhibit different strategies for predator avoidance. Locusts rely primarily on walking and jumping to evade predators, whereas stick insects become cataleptic, catalepsy forming a major component of the twig mimesis exhibited by this species. The neuronal networks that control postural leg movements in locusts and stick insects are tuned differently to their specific behavioral tasks. An important prerequisite for the production of catalepsy in the stick insect is the marked velocity dependency of the control network, which appears to be generated at the level of nonspiking local interneurons. We examined interneuronal pathways in the network controlling the femur-tibia joint of the locust middle leg and compared its properties with those described for the stick insect middle leg. It was our aim to identify possible neural correlates of the species-specific behavior with regard to postural leg motor control. 2. We obtained evidence that the neuronal networks that control the femur-tibia joints in the two species consist of morphologically and physiologically similar--and thus probably homologous--interneurons. Qualitatively, these interneurons receive the same input from the femoral chordotonal organ receptors and they drive the same pools of leg motoneurons in both species. 3. Pathways that contribute to the control of the femur-tibia joint include interneurons that support both "resisting" and "assisting" responses with respect to the motoneuron activity that is actually elicited during reflex movements. Signal processing via parallel, antagonistic pathways therefore appears to be a common principle in insect leg motor control. 4. Differences between the two insect species were found with regard to the processing of velocity information provided by the femoral chordotonal organ. Interneuronal pathways are sensitive to stimulus velocity in both species. However, in the locust there is no marked velocity dependency of the interneuronal responses, whereas in the same interneurons of the stick insect it is pronounced. This characteristic was maintained at the level of the motoneurons controlling the femur-tibia joint. Pathways for postural leg motor control in the locust thus lack an important prerequisite for the generation of catalepsy, that is, a marked velocity dependency.

Animals↗
PubMed1995

Nonspiking local interneurons in insect leg motor control. II. Role of nonspiking local interneurons in the control of leg swing during walking.

1. Nonspiking local interneurons (NSIs) were recorded intracellularly in the mesothoracic ganglion of semi-intact locusts walking on a treadwheel. Interneurons were characterized by their connectivity to motoneurons. Their activity patterns in the step cycle and the effect current injection had on the leg movement were analyzed. We examined interneurons that provided excitatory or inhibitory synaptic drive to a subset of motoneurons active during the swing movement of walking. 2. Interneuron activity was observed to support or oppose the actual leg movement. Both supporting and opposing interneurons were active simultaneously, lending support to the idea that the actual motor output of walking is generated by the adjustment of parallel antagonistic pathways of signal processing. 3. The examined interneurons showed qualitatively the same patterns of activity during forward and backward walking. This indicates that swing movement in both situations may be generated by similar neuronal networks (although the mechanism of movement reversal remains unclear). 4. At least two functional types of NSIs could be distinguished. First, there were interneurons whose depolarization patterns showed distinct variability, often correlated with duration or amplitude of the swing movement. As a rule, current injection had minor, if any, effects on leg movement. Populations of these interneurons appear to be involved in the control of a coordinated swing movement by driving appropriate sets of muscle groups. The second type of NSIs showed more stereotyped activity patterns that varied relatively little with changes in the swing movement. Current injection had strong effects on the leg movement and could, for example, arrest the leg in the stance phase. These interneurons appear to be primarily involved in the trigger mechanism of leg swing.

Animals↗
PubMed1995

Chronic granulomatous disease assessed by single-cell granulocyte oxidative burst activity.

Here we report on a case of chronic granulomatous disease (CGD) in a 3-year-old boy who suffered from severe repeated bacterial infections including multiple liver abscesses. The case is of interest because (1) the disease is very rare (it is the first case of CGD diagnosed at the Clinic for Pediatric Medicine, University of Innsbruck), (2) the diagnosis, based on clinical parameters and the nitrobluetetrazolium test was completed and validated by single-cell measurements of respiratory-burst activity of the patient's granulocytes in a fluorescence-activated cell sorter (FACS), and (3) the applied FACS method, adapted in our laboratory, presents one of the most sensitive and reliable methods to evaluate this aspect of disturbed granulocyte function.

Cell Separation↗
PubMed1995

Features of oral immunization.

In this review, we focus on some key areas concerning the unique properties of the mucosal immune system. They are: (1) the fact that the common mucosal immune system consists of different compartments; (2) the advantages of oral vaccination, which can be exploited to antigen-specific sIgA-mediated local immune responses as well as systemic immunity; (3) efficacious oral immunization against respiratory infections; (4) oral tolerance with respect to activation of T cells which, after declining, can be repeatedly reinduced without changes in profile or magnitude, and (5) the use of transgenic plants as a new vaccine source for a new vaccination strategy, i.e. employing edible dietary vaccines.

Administration, Oral↗
PubMed1995

[DACS--a cost-advantageous nuclear medical document archiving and communication system].

A simple and inexpensive solution for the complete digital management of a nuclear medical institution is presented. The starting point was provided by five different gamma cameras, one of which was equipped with a PC-based ICON computer. The remaining cameras were first adapted to ICONs and subsequently integrated into a Macintosh LAN, of 15 computers, allowing the entire image acquisition, processing, interpretation, documentation and archiving to be performed on the same comfortable menu-guided surface. Complete patient management is accomplished by a specific commercially available data base to which a specially developed image and document archive was connected. The system described provides complete digital management without the conventional filing systems at the moderate price of approximately DM 11 x 10(4) plus the cost of the new camera computers.

Computer Graphics↗
PubMed1995

Proprioceptive sensory neurons of a locust leg receive rhythmic presynpatic inhibition during walking.

Mechanosensory neurons from a proprioceptor (the femoral chordotonal organ) signal the movements and positions of the femorotibial joint of a locust leg. Intracellular recordings from these neurons during walking show that their spikes are superimposed on a depolarizing synaptic input generated near their output terminals in the CNS. The depolarization consists of a rhythmic synaptic input at each step, and a sustained input that begins before walking commences. In different sensory neurons, which signal particular features of the movement, the rhythmic depolarization occurs at distinct times during either the swing or stance phases of the step cycle. The depolarizing input is timed to coincide with the greatest spike response of a sensory neuron. The input is associated with a conductance change, appears to reverse just above resting potential, and thus has similar properties to the presynaptic inhibition in these same neurons during imposed joint movements (Burrows and Laurent, 1993; Burrows and Matheson, 1994). Three sources could contribute to these inputs: (1) interactions between sensory neurons of the same receptor signaling the same movement, (2) signals from different receptors in the same leg and other legs, and (3) outputs of central neurons involved in generating walking. When the leg, whose movements the sensory neurons signal is removed, both the sustained and rhythmic synaptic inputs persist. Sensory neurons in isolated ganglia treated with pilocarpine are also depolarized in phase with a rhythmic pattern expressed in leg motor neurons, indicating that central neurons must contribute. The maintained synaptic input to the terminals means that the overall effectiveness of the sensory spikes in evoking EPSPs in postsynaptic neurons will be reduced during walking, and the rhythmic component means that the spikes from particular sensory neurons will be further reduced at particular phases of the step cycle that they signal best.

Animals↗
PubMed1995

First EBV vaccine trial in humans using recombinant vaccinia virus expressing the major membrane antigen.

In the absence of a truly representative animal model, the question of whether EBV-related diseases can be prevented by a vaccine has been studied for the first time in humans. A live recombinant virus based on the licensed vaccinia strain Tien Tan, expressing under the 11K vaccinia promoter the major EBV membrane antigen BNLF-1 MA (gp 220-340), was constructed and tested in three different human populations: EBV-positive and vaccinia-virus-exposed adults; EBV-positive, non-vaccinia-virus-exposed juveniles; and EBV and vaccinia virus-naive infants. No significant titre variations for EBV were observed in the adults, but EBV-neutralising titres increased in the vaccinated juveniles, while antibodies to VCA of EBV remained unchanged. All nine vaccinated infants developed antibodies to MA (membrane antigen) with neutralising properties in vitro; three of these infants were infected by EBV via natural routes over a period of 16 months after vaccination and all ten unvaccinated control infants became infected. It has been shown for the first time that protection against and/or delay of EBV infection by the natural route is possible in humans and that live vaccinia vectors can be used and are efficacious.

Adult↗
PubMed1995

[Eradication of Legionella from warm water systems--documentation of our own experiences with thermal disinfection].

A nosocomial case of Legionellosis in a recently built hospital was the reason for an investigation and thermal disinfection of the complete warm water distribution system. Furthermore weak points in the tubing of the warm water system, which promoted the contamination of potable water, were eliminated as far as technically possible. A lasting reduction of the numbers of Legionella spp. isolated could be measured (factor 10-1000), but a complete decontamination of the warm water distribution system was not possible. As Legionella spp. may cause serious infections in immunodeficient patients, additional measures for the disinfection of the hospital water distribution system must be taken into consideration.

Cross Infection↗
PubMed1995

Strategy for typing human papillomaviruses by RFLP analysis of PCR products and subsequent hybridization with a generic probe.

Genital human papillomaviruses (HPV) were detected by PCR using L1 consensus primers MY09 and MY11. To determine the underlying HPV type(s). PCR products were subsequently analyzed employing a combination of restriction fragment length polymorphism (RFLP) and hybridization with a generic oligonucleotide probe that binds to a conserved region located close to the MY11-binding site within the PCR products. Using computer-assisted sequence analysis, the lengths of the corresponding BamHI, DdeI, HaeIII, HinfI and PstI restriction fragments hybridizing with the generic probe were calculated, revealing distinct patterns for each of the 45 mucosal HPV types. This method is superior to RFLP analysis since it is not impaired by large amounts of restriction fragments resulting from nonspecific PCR products. Moreover, considering clinical specimens containing two or three different HPV types, direct sequencing of PCR products will be inconclusive, and the increased number of restriction fragments will complicate interpretation of RFLP patterns. Subsequent hybridization with the generic probe, however, results in the appearance of, at most, 2 or 3 bands per restriction enzyme digest and thus facilitates identification of the underlying HPV types.

Anal Canal↗
PubMed1994

A flow cytometric long-term cytotoxicity assay.

A method to evaluate cytotoxic effects applicable over a wide range of incubation times has been developed. It is based on quantification by flow cytometry of dead and viable target cells stained by covalently binding the fluorescent dye fluorescein isothiocyanate (FITC). The staining with FITC did not affect cell viability and growth parameters and was stable enough to identify target cells for at least 2 days. The flow cytometric analysis of the cell mixture was performed in a test system with activated CD8+ lymphocytes as effector cells and melanoma M21 cells as targets in the presence of appropriate bispecific antibodies and revealed a rather complex pattern composed of several distinct cell subsets which were identified by use of antibodies to lymphocyte antigens. The assay compared favourably with results from a conventional 51Cr release assay obtained after 4 h and 8 h of incubation and from a target cell adherence assay obtained after 24 h of incubation. The application of the method described herein is especially advantageous for the evaluation of long-term cytotoxic effects. Furthermore, it provides valuable multi-parameter information which is useful for elucidating mechanisms of cytotoxicity.

Animals↗
PubMed1994

The Epstein-Barr virus-encoded glycoprotein gp 110 (BALF 4) can serve as a target for antibody-dependent cell-mediated cytotoxicity (ADCC).

Antibody-dependent cell-mediated cytotoxicity (ADCC) is thought to play a major role in controlling the spread of the Epstein-Barr virus (EBV) in an infected individual. Recently, the viral membrane protein gp 350/220, which is also expressed at the surface of the virus producing cell, was identified as a target for ADCC reactions. Due to its glycoprotein nature, the EBV protein gp 110 is another possible ADCC target. It is one of the most abundant proteins found during the late phase of viral replication; until now, however, researchers have not been able to localize it on the surface of EBV positive cells. By means of recombinant vaccinia viruses containing the genes for gp 350/220 and gp 110, respectively, we expressed these proteins in lymphoblastoid cells, which were then used as targets in ADCC studies with sera from EBV-positive and -negative individuals. In these experiments we were able to demonstrate the feasibility of our approach for the investigation of EBV-specific ADCC reactions and could confirm the role of gp 350/220 as an ADCC target. Furthermore, we were able to show that gp 110 can also be recognized in an ADCC reaction, proving that at least some gp 110 molecules must be expressed at the cell surface.

Antibody-Dependent Cell Cytotoxicity↗
PubMed1994

A simple and sensitive method to study effects mediated by soluble lymphokines as demonstrated by the interaction of CD4+ and CD8+ cell subsets during T cell activation.

A method is described for the study of lymphokine-mediated cellular interactions using triple wells, which permits co-culture of cell subpopulations without direct physical contact. The triple wells are constructed by slitting the walls to half height between three adjacent wells of a 96-well microtiter plate. The cells under study are positioned in the outer two wells, whereas the middle well serves to separate the cells. The half slits permit the wells to be treated independently before filling the triple well with the culture medium and prevents cell leakage thereafter. The feasibility of the method was established by studying the interaction of isolated CD4+ and CD8+ T cell subsets during T cell proliferation induced by immobilized anti-CD3 and anti-CD28 monoclonal antibodies.

CD4-Positive T-Lymphocytes↗
PubMed1994

Blood group ABO-related glycosylation of urothelial cell lines: immunocytological, enzymatic, and genetic characterization.

Three immortalized, human urothelial cell lines were characterized with respect to their ABO-related carbohydrate phenotypes using a panel of monoclonal antibodies directed to a series of carbohydrate epitopes (Lac, sialylated Lac, Le(a), sialylated Le(a), Le(x), sialylated Le(x), H types I and II, Ley, Leb, A monofucosylated types I and II, ALey, Aleb, and A type III). The glycosyltransferases forming some of these epitopes (beta 1-3/4 galactosyltransferase, alpha 1-2 fucosyltransferase, alpha 1-3 galactosyltransferase, and alpha 1-3-N-acetyl-galactosaminyltransferase) were determined by enzyme assays. The ABO gene complex was analyzed by Southern blotting, Northern blotting, and polymerase chain reaction across the O deletion and across base differences between the A and B alleles. The immunocytochemical stainings showed marked differences between the three cell lines; the high grade (tumorigenic, metastatic) cell line showed difucosylated types I and II structures, and the low grade (nontumorigenic, nonmetastatic) cell lines showed monofucosylated types I and II structures. Polymerase chain reaction genotyping of the cell lines indicated that one was OO, one was AA, and one was A plus a mutated allele. Northern blotting showed RNA encoding the A transferase. However, even though both of the A cell lines seemed to have an intact gene, which could produce A transferase and transcribed RNA, none of them showed any activity of the A gene encoded enzyme or any A-structures at the cell surface. In contrast, the three other examined glycosyltransferases were active. The three urothelial cell lines reflect in vivo findings in humans. They represent a competent system for in vitro studies of the different carbohydrate transferase genes responsible for the carbohydrate structures expressed on the cell surface in bladder tumors.

ABO Blood-Group System↗