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Biomedical subjects

S Weiss

Publications and source records attributed to S Weiss.

At least 181 records · Page 10Linked to original sources
PubMed1997

Membrane specific mapping and colocalization of malarial and host skeletal proteins in the Plasmodium falciparum infected erythrocyte by dual-color near-field scanning optical microscopy.

Accurate localization of proteins within the substructure of cells and cellular organelles enables better understanding of structure-function relationships, including elucidation of protein-protein interactions. We describe the use of a near-field scanning optical microscope (NSOM) to simultaneously map and detect colocalized proteins within a cell, with superresolution. The system we elected to study was that of human red blood cells invaded by the human malaria parasite Plasmodium falciparum. During intraerythrocytic growth, the parasite expresses proteins that are transported to the erythrocyte cell membrane. Association of parasite proteins with host skeletal proteins leads to modification of the erythrocyte membrane. We report on colocalization studies of parasite proteins with an erythrocyte skeletal protein. Host and parasite proteins were selectively labeled in indirect immunofluorescence antibody assays. Simultaneous dual-color excitation and detection with NSOM provided fluorescence maps together with topography of the cell membrane with subwavelength (100 nm) resolution. Colocalization studies with laser scanning confocal microscopy provided lower resolution (310 nm) fluorescence maps of cross sections through the cell. Because the two excitation colors shared the exact same near-field aperture, the two fluorescence images were acquired in perfect, pixel-by-pixel registry, free from chromatic aberrations, which contaminate laser scanning confocal microscopy measurements. Colocalization studies of the protein pairs of mature parasite-infected erythrocyte surface antigen (MESA) (parasite)/protein4.1(host) and P. falciparum histidine rich protein (PfHRP1) (parasite)/protein4.1(host) showed good real-space correlation for the MESA/protein4.1 pair, but relatively poor correlation for the PfHRP1/protein4.1 pair. These data imply that NSOM provides high resolution information on in situ interactions between proteins in biological membranes. This method of detecting colocalization of proteins in cellular structures may have general applicability in many areas of current biological research.

Cells, Cultured↗
PubMed1997

TAP-dependent major histocompatibility complex class I presentation of soluble proteins using listeriolysin.

Immunization of mice with mixtures of listeriolysin, a pore-forming hemolysin secreted by the pathogenic bacterium Listeria monocytogenes, together with soluble ovalbumin, nucleoprotein of influenza virus, or beta-galactosidase of Escherichia coli, resulted in strong cytotoxic CD8 T cell responses to each of the respective passenger proteins in vivo. Also, the concomitant addition of either protein with listeriolysin to target cells elicited efficient sensitization of these cells which could be attributed to the pore-forming activity of listeriolysin. This response was dependent upon a functional TAP transporter and was inhibitable by brefeldin A, indicating the transfer of the soluble proteins into the cytosol and the classical major histocompatibility (MHC) class I presentation pathway. The treatment of target cells with listeriolysin under our experimental conditions did not affect cell viability and the pores generated by listeriolysin treatment were repaired within 60 min. Introduction of soluble proteins into the MHC class I presentation pathway by listeriolysin provides a powerful system to study the cytotoxic response towards intracellular pathogens and would allow for rapid screening of potential antigens in vaccine formulations.

ATP-Binding Cassette Transporters↗
PubMed1997

Excessive degradation of intracellular protein in macrophages prevents presentation in the context of major histocompatibility complex class II molecules.

The endogenous major histocompatibility complex (MHC) class II presentation pathway allows biosynthesized, intracellular antigens access for presentation to MHC class II-restricted T cells. This pathway has been well documented in B cells and fibroblasts, but may not be universally available in all antigen-presenting cell types. This study compares the ability of different antigen-presenting cells, expressing endogenous C5 protein (fifth component of mouse complement) as a result of transfection, to present their biosynthesized C5 to MHC class II-restricted T cells. B cells and fibroblasts expressing C5 were able to present several epitopes of this protein with MHC class II molecules, whereas macrophages were unable to do so, but readily presented C5 from an extracellular source. However, macrophage presentation of endogenous C5 could be achieved when they were treated with low doses of the lysosomotropic agent ammonium chloride. In the presence of an inhibitor of autophagy, presentation of endogenous C5 was abrogated, indicating that biosynthesized C5 is shuttled into lysosomal compartments for degradation before making contact with MHC class II molecules. Taken together, this suggests that proteolytic activity in lysosomes of macrophages may be excessive, compared with fibroblasts and B cells, and destroys epitopes of the C5 protein before they can gain access to MHC class II molecules. Thus, there are inherent differences in presentation pathways between antigen-presenting cell types; this could reflect their specialized functions within the immune system with macrophages focussing preferentially on internalization, degradation, and presentation of extracellular material.

Adenine↗
PubMed1997

Antigen-specific T cell receptor antagonism by antigen-presenting cells treated with the hemolysin of Listeria monocytogenes: a novel type of immune escape.

We have examined the influence of listeriolysin O (LLO), the hemolysin secreted by the pathogenic bacterium Listeria monocytogenes, on major histocompatibility complex class II-dependent T cell activation. Stimulation of T cells by native antigens but not by peptides is inhibited upon pretreatment of antigen-presenting cells (APC) with LLO. Experiments presented here reveal that this inhibition is not due to a lack in processing of antigen by APC but is the result of an irreversible inactivation of T cells that recognize antigen on LLO-treated APC. Incubation of mixtures of two different T cells where only one antigen was presented on LLO-treated APC suggested that T cell inactivation is antigen specific. The inactivation was dominant and could be observed even in the presence of amounts of synthetic peptides that normally lead to T cell responses. This condition is reminiscent of the T cell inhibition observed when antagonistic and stimulatory peptides are added to APC at the same time. Our results thus reveal a novel type of interference by pathogens with antigen presentation and T cell stimulation that could give the pathogen a decisive advantage in dissemination and disease.

Amino Acid Sequence↗
PubMed1997

Receptive field dimensions of lateral geniculate cells in the common marmoset (Callithrix jacchus).

We measured the spatial receptive field dimensions of cells in the lateral geniculate nucleus (LGN) of the common marmoset (Callithrix jacchus) using a bipartite field stimulus in which the two halves of the field were modulated identically but in counterphase. Horizontal and vertical edges between the two fields were positioned at different locations in the receptive field. By assuming that centers and surrounds have gaussian profiles, we were able to obtain a satisfactory mathematical description of the data. Receptive field centers were about a factor 1.6 larger than those of macaque LGN cells, in accordance with the smaller marmoset eye. There was a limited correspondence with dendritic tree dimensions of marmoset retinal ganglion cells. We further found that center and surround gaussians were not always concentric, and that the centers of some cells were elongated. This might allow some direction or orientation biases in LGN cells.

Algorithms↗
PubMed1997

Temporal properties of marmoset lateral geniculate cells.

We measured the temporal modulation transfer functions (TMTFs) of cells in the marmoset lateral geniculate nucleus (LGN) at three different luminance levels, and described the responses with a linear model. It was found that qualitatively there are many similarities with the temporal response properties of macaque and marmoset retinal ganglion cells. M-cells displayed stronger attenuation at lower temporal frequencies, and showed more nonlinearities (such as saturation and a contrast gain control) than P-cells. We therefore propose that the temporal properties of the visual system of New and Old World monkeys are similar at least up to the LGN. However, there are some quantitative differences, indicating that response alterations take place at the stage of synaptic transmission in the LGN. The most important are an attenuation of the responses to higher temporal frequencies and the smaller differences between parvo- and magnocellular cell responsivities. Cell responses to square-wave modulation were also measured and compared with predictions from a linear systems analysis. The linear systems analysis gave reasonable predicted responses to square-wave modulation, but these predictions were poor than those for retinal ganglion cells, indicating that additional nonlinearities are introduced at the synaptic transition in the LGN.

Animals↗
PubMed1997

The human 37-kDa laminin receptor precursor interacts with the prion protein in eukaryotic cells.

Prions are thought to consist of infectious proteins that cause transmissible spongiform encephalopathies. According to overwhelming evidence, the pathogenic prion protein PrPSc converts its host encoded isoform PrPC into insoluble aggregates of PrPSc, concomitant with pathological modifications (for review, see refs. 1-3). Although the physiological role of PrPC is poorly understood, studies with PrP knockout mice demonstrated that PrPC is required for the development of prion diseases. Using the yeast two-hybrid technology in Saccharomyces cerevisiae, we identified the 37-kDa laminin receptor precursor (LRP) as interacting with the cellular prion protein PrPC. Mapping analysis of the LRP-PrP interaction site in S. cerevisiae revealed that PrP and laminin share the same binding domain (amino acids 161 to 180) on LRP. The LRP-PrP interaction was confirmed in vivo in insect (Sf9) and mammalian cells (COS-7). The LRP level was increased in scrapie-infected murine N2a cells and in brain and spleen of scrapie-infected mice. In contrast, the LRP concentration was not significantly altered in these organs from mice infected with the bovine spongiform encephalopathic agent (BSE), which have a lower PrPSc accumulation. LRP levels, however, were dramatically increased in brain and pancreas, slightly increased in the spleen and not altered in the liver of crapie-infected hamsters. These data show that enhanced LRP concentrations are correlated with PrPSc accumulation in organs from mice and hamsters. The laminin receptor precursor, which is highly conserved among mammals and is located on the cell surface, may act as a receptor or co-receptor for the prion protein on mammalian cells.

Actins↗
PubMed1997

Listeriolysin and IrpA are major protein targets of the human humoral response against Listeria monocytogenes.

We have examined the human humoral immune response directed against proteins of Listeria monocytogenes in both healthy individuals and listeriosis patients. Two major targets for an antibody response were found in individuals that did not suffer from listeriosis: listeriolysin (Hly) and the recently described internalin-related protein (IrpA). In contrast, the humoral response in listeriosis patients appears to be more heterogeneous and included Hly, IrpA, InlB, and ActA as major targets.

Antibodies, Bacterial↗
PubMed1997

RNA aptamers specifically interact with the prion protein PrP.

We have isolated RNA aptamers which are directed against the recombinant Syrian golden hamster prion protein rPrP23-231 (rPrPc) fused to glutathione S-transferase (GST). The aptamers did not recognize the fusion partner GST or the fusion protein GST::rPrP90-231 (rPrP27-30), which lacks 67 amino acids from the PrP N terminus. The aptamer-interacting region of PrPc was mapped to the N-terminal amino acids 23 to 52. Sequence analyses suggest that the RNA aptamers may fold into G-quartet-containing structural elements. Replacement of the G residues in the G quartet scaffold with uridine residues destroyed binding to PrP completely, strongly suggesting that the G quartet motif is essential for PrP recognition. Individual RNA aptamers interact specifically with prion protein in brain homogenates from wild-type mice (C57BL/6), hamsters (Syrian golden), and cattle as shown by supershifts obtained in the presence of anti-PrP antibodies. No interaction was observed with brain homogenates from PrP knockout mice (prn-p(0/0)). Specificity of the aptamer-PrP interaction was further confirmed by binding assays with antisense aptamer RNA or a mutant aptamer in which the guanosine residues in the G tetrad scaffold were replaced by uridine residues. The aptamers did not recognize PrP27-30 in brain homogenates from scrapie-infected mice. RNA aptamers may provide a first milestone in the development of a diagnostic assay for the detection of transmissible spongiform encephalopathies.

Animals↗
PubMed1997

Homozygosity by descent for a rare mutation in the myophosphorylase gene is associated with variable phenotypes in a Druze family with McArdle disease.

We examined a large consanguineous Druze family with McArdle disease for mutations in the glycogen myophosphorylase (PYGM) gene. All affected subjects were autozygous for a single G to A transition that abolishes the 5' consensus splice site in the first nucleotide of intron 14. The G to A transition is a rare mutation, with only one previous report in a single white subject heterozygous for this mutation and another, more common, mutation at codon 49. The kindred in our study is the first family reported in which disease is caused by homozygosity for this rare mutation. This kindred was originally reported as the first familial case of McArdle disease in the Druze.

Cell Line, Transformed↗
PubMed1997

The development of subretinal microphotodiodes for replacement of degenerated photoreceptors.

There are presently several concepts to restore vision in blind or highly visually handicapped persons by implanting electronic devices into the eye in order to partially restore vision. Here, the approach to replace retinal photoreceptors by a subretinally implanted microphotodiode array (MPDA) is summarized. A survey is given on the present state of the development of MPDAs, the possibility of in vitro and in vivo tests as well as first results on biocompatibility and histology. Additionally, electrophysiological recordings in rabbits and rats are presented which have received such subretinal implants.

Animals↗
PubMed1997

[Morphologic parameters for quantitative determination of inflammatory activity of the peritoneum].

The morphology of the inflammatory activity of the peritoneum has been measured qualitatively but quantitative assessments are not common. In a standardized rat model we induced chronic abscess-forming peritonitis after laparotomy and inoculation of 2 ml Bacteroides fragilis suspension at a concentration of 10(9)/ml colony-forming units. The morphological inflammatory activity was determined quantitatively by staining the specimen of the peritoneum with naphthol-AS-D-chloracetate-esterase (NASDCE); through this staining the cytoplasm of granulocytes and tissue mast cells were marked. The peritonitis group (n = 53) and controls (n = 15) were randomly divided into three subgroups (nPeritonitis = 17/18/18 vs. ncontrol = 5/5/5) and observed for 3/7/14 days, respectively. On days 3/7/14 we diagnosed intra-abdominal abscesses in 2 of 17, 13 of 18, and 12 of 18 animals in the peritonitis group. In controls there were no abscesses (P < 0.05). The total cellularity and NASDCE-positive rates on days 3/7/14 in the peritonitis group were 301/409/280 (vs. 155/240/273 in controls) and 1.8/2.9/3.6% (vs. 0.7/0.9/1.4%) in the non-abscess-forming regions and 392/661/625 and 14.4/12.9/11.5% in the abscess-surrounding regions in the infected animals, respectively (P < 0.05). We conclude that the qualitative histological evidence of the morphological inflammatory activity of the peritoneum in the form of an abscess can be supplemented by a quantitative method. Through NASDCE staining the granulocyte and tissue mast cell proportion of the total cellularity as main indicators of the local inflammatory activity can be estimated in peritonitis. This method can be helpful in deciding when to definitively close the abdomen in the course of a programmed lavage treatment in peritonitis.

Abdominal Abscess↗
PubMed1997

[Urgent need for prevention of alcohol drinking among Arab youth].

A study was conducted in the winter of 1996 among 2,220 Arab adolescents in northern Israel. This fourth study among Arab youth dealt with frequency of drinking and amounts of alcohol drunk during a drinking bout. Among Christian, Druze and Moslem males there were 81.72, 49.61% and 31.93%, respectively, who drank. The figures for females were 36.75%, 11.25% and 12.78%. About 4 % of Moslem males drank 5 drinks or more consecutively, daily or every 2-3 days. The need for preventive efforts in the Arab sector is stressed.

Adolescent↗
PubMed1996

Desmoid fibromatosis of the sinonasal tract and nasopharynx. A clinicopathologic study of 25 cases.

BACKGROUND: Desmoid fibromatoses are a group of nonmetastasizing, well differentiated, unencapsulated fibrous tissue proliferations that have a tendency for local invasion and recurrence. Biologically, they fall in an intermediate category between benign fibrous lesions and fibrosarcoma. Because of the rarity of this lesion in the upper respiratory tract and inadequate characterization of its biologic potential in the literature, this study was undertaken. METHODS: The files of the Armed Forces Institute of Pathology were reviewed for cases of fibromatosis involving the sinonasal and nasopharyngeal areas that occurred between 1885 and 1985. For each case, histologic materials were reviewed, clinical data tabulated, and follow-up obtained. RESULTS: Twenty-five cases were identified. Sixteen patients were male and 9 female, ranging in age from 8 months to 62 years (mean, 29 years and 11 months). A single site was involved in 18 patients and multiple contiguous adjacent sites in 7. The maxillary sinus was the site most frequently involved (22 patients), followed by the nasal cavity (5 patients), the ethmoid sinus (4 patients), orbit (4 patients), sphenoid and frontal sinuses (2 patients each), and the nasopharynx (1 patient). Twenty-four patients were followed for periods ranging from 1 year to 20 years and 7 months (median, 6 years and 9 months; mean, 8 years and 2 months). At last follow-up, 18 patients were alive and well with no evidence of disease, 2 patients were alive with unknown disease histories, and 3 patients were alive with recurrent or residual disease. One patient died without evidence of disease. Five patients (21%) (4 adults and 1 child) developed recurrences; 3 patients had 1 recurrence at 6, 16, and 34 months, respectively, 1 patient had 2 recurrences at 3.5 and 5.5 months, and 1 patient was alive with recurrent disease at 6.5 years. One patient was lost to follow-up. CONCLUSIONS: Twenty-five cases of desmoid fibromatosis involving the sinonasal tract and nasopharynx were described. These lesions appear to have lower recurrence rates and morbidity than desmoid fibromatoses arising in many other areas of the body.

Adult↗
PubMed1996

Multipotent CNS stem cells are present in the adult mammalian spinal cord and ventricular neuroaxis.

Neural stem cells in the lateral ventricles of the adult mouse CNS participate in repopulation of forebrain structures in vivo and are amenable to in vitro expansion by epidermal growth factor (EGF). There have been no reports of stem cells in more caudal brain regions or in the spinal cord of adult mammals. In this study we found that although ineffective alone, EGF and basic fibroblast growth factor (bFGF) cooperated to induce the proliferation, self-renewal, and expansion of neural stem cells isolated from the adult mouse thoracic spinal cord. The proliferating stem cells, in both primary culture and secondary expanded clones, formed spheres of undifferentiated cells that were induced to differentiate into neurons, astrocytes, and oligodendrocytes. Neural stem cells, whose proliferation was dependent on EGF+bFGF, were also isolated from the lumbar/sacral segment of the spinal cord as well as the third and fourth ventricles (but not adjacent brain parenchyma). Although all of the stem cells examined were similarly multipotent and expandable, quantitative analyses demonstrated that the lateral ventricles (EGF-dependent) and lumbar/sacral spinal cord (EGF+bFGF-dependent) yielded the greatest number of these cells. Thus, the spinal cord and the entire ventricular neuroaxis of the adult mammalian CNS contain multipotent stem cells, present at variable frequency and with unique in vitro activation requirements.

Animals↗
PubMed1996

Expression of a second receptor rescues self-specific T cells from thymic deletion and allows activation of autoreactive effector function.

Allelic exclusion at the T-cell receptor alpha chain locus is incomplete resulting in the generation of T cells that express two T-cell receptors. The potential involvement of such T cells in autoimmunity has been suggested [Padovan, E., Casorati, G., Dellabona, P., Meyer, S., Brockhaus, M. & Lanzavecchia, A. (1993) Science 262, 422-424; Heath, W. R. & Miller, J. F. A. P. (1993) J. Exp. Med. 178, 1807-1811]. Here we show that expression of a second T-cell receptor can rescue T cells with autospecific receptors from thymic deletion and allow their exit into the periphery. Dual receptor T cells, created by constitutive expression of two transgenic T-cell receptors on a Rag1-/- background, are tolerant to self by maintaining low levels of autospecific receptor, but selfreactive effector function (killing) can be induced through activation via the second receptor. This opens the possibility that T cells carrying two receptors in the periphery of normal individuals contain putatively autoreactive cells that could engage in autoimmune effector functions after recognition of an unrelated environmental antigen.

Animals↗