Biomedical subjects

R Vogels

Publications and source records attributed to R Vogels.

At least 19 recordsLinked to original sources

PubMed2002

Exploiting the natural diversity in adenovirus tropism for therapy and prevention of disease.

Since targeting of recombinant adenovirus vectors to defined cell types in vivo is a major challenge in gene therapy and vaccinology, we explored the natural diversity in human adenovirus tissue tropism. Hereto, we constructed a library of Ad5 vectors carrying fibers from other human serotypes. From this library, we identified vectors that efficiently infect human cells that are important for diverse gene therapy approaches and for induction of immunity. For several medical applications (prenatal diagnosis, artificial bone, vaccination, and cardiovascular disease), we demonstrate the applicability of these novel vectors. In addition, screening cell types derived from different species revealed that cellular receptors for human subgroup B adenoviruses are not conserved between rodents and primates. These results provide a rationale for utilizing elements of human adenovirus serotypes to generate chimeric vectors that improve our knowledge concerning adenovirus biology and widen the therapeutic window for vaccination and many different gene transfer applications.

Adenoviruses, Human↗

PubMed2001

Treatment of malignant gliomas with a replicating adenoviral vector expressing herpes simplex virus-thymidine kinase.

We evaluated the interaction between oncolytic, replication-competent adenoviral vectors and the herpes simplex virus-1 thymidine kinase (HSV1-tk) gene/ganciclovir (GCV) suicide system for the treatment of malignant gliomas. We constructed a panel of replication-competent adenoviral vectors in which the luciferase (IG.Ad5E1(+). E3Luc) or HSV1-tk gene (IG.Ad5E1(+).E3TK) replace the M(r) 19,000 glycoprotein (gp19K) coding sequence in the E3 region. IG.Ad5E1. IG.Ad5.ClipLuc and IG.AdApt.TK are E1-deleted viruses that contain the luciferase or the HSV1-tk gene in the former E1 region driven by the human cytomegalovirus promoter. IG.Ad5. Sarcoma 1800HSA.E3Luc contains an irrelevant gene in the E1 region, whereas the gp19K coding sequence in the E3 region is replaced by the luciferase gene as in the replicating virus IG.Ad5E1(+).E3Luc. For in vitro experiments, we used a panel of human glioma cell lines (U87 MG, T98G, A172, LW5, and U251), a rat gliosarcoma cell line (9 L), and human lung (A549) and prostate carcinoma (P3) cell lines. In vitro, GCV sensitivity (10 microg/ml) was studied in U87 MG cells after infection at a multiplicity of infection of 1 and 10. A s.c. U87 MG glioma xenograft model was established in NIH-bg-nu-xid mice. Tumors of 100-150 mm(3) were treated with a single injection of adenovirus 10(9) IU suspended in 100 microl of PBS, and GCV 100 mg/kg was administered i.p. twice daily for 7 days. The cytopathic effect of all three replication-competent adenoviral vectors was similar to the cytopathic effect of wild-type adenovirus 5 on all human cell lines tested, indicating that deletion of the E3 gp19K sequences did not affect the oncolytic effect of the vectors. In vitro, luciferase expression was the same for both E1-deleted vectors (IG.Ad5.ClipLuc and IG.Ad5. Sarcoma 1800HSA.E3Luc), demonstrating the strength of the internal E3 promoter even in the absence of E1A. However, in vitro expression levels obtained with replication-competent IG.Ad5E1(+). E3Luc were 3 log higher (allowing infection with a 2-3-log lower multiplicity of infection) in the human cell lines. In U87 MG glioma cells, the oncolytic effect of replication-competent IG.Ad5E1(+).E3TK was significantly enhanced by the addition of GCV and greatly exceeded the cytotoxicity of replication-incompetent IG.AdApt.TK combined with GCV. In established s.c. U87 MG glioma xenografts, a single injection of IG.Ad5E1(+).E3TK resulted in a significant slowing of tumor growth and prolonged survival compared with injection of IG.AdApt.TK. Addition of GCV slowed tumor growth, further adding to survival. In conclusion, the oncolytic effect of replicating adenoviral vectors and HSV1-tk/GCV have potent antitumor effects in gliomas. When combined, these two approaches are complementary, resulting in a significantly improved treatment outcome. In addition, replication-competent adenoviral vectors missing the E3 gp19K coding sequences, have oncolytic efficacy comparable with wild type. In combination with high expression levels obtained with the natural E3 promoter, such vectors are promising new anticancer agents.

Adenoviridae↗

PubMed2001

Macaque inferior temporal neurons are selective for three-dimensional boundaries and surfaces.

The lower bank of the superior temporal sulcus (TEs), part of the inferior temporal cortex, contains neurons selective for disparity-defined three-dimensional (3-D) shape. The large majority of these TEs neurons respond to the spatial variation of disparity, i.e., are higher-order disparity selective. To determine whether curved boundaries or curved surfaces by themselves are sufficient to elicit 3-D shape selectivity, we recorded the responses of single higher-order disparity-selective TEs neurons to concave and convex 3-D shapes in which the disparity varied either along the boundary of the shape, or only along its surface. For a majority of neurons, a 3-D boundary was sufficient for 3-D shape selectivity. At least as many neurons responded selectively to 3-D surfaces, and a number of neurons exhibited both surface and boundary selectivity. The second aim of this study was to determine whether TEs neurons can represent differences in second-order disparities along the horizontal axis. The results revealed that TEs neurons can also be selective for horizontal 3-D shapes and can code the direction of curvature (vertical or horizontal). Thus, TEs neurons represent both boundaries and surfaces curved in depth and can signal the direction of curvature along a surface. These results show that TEs neurons use not only boundary but also surface information to encode 3-D shape properties.

Animals↗

PubMed2001

Practising orientation identification improves orientation coding in V1 neurons.

The adult brain shows remarkable plasticity, as demonstrated by the improvement in fine sensorial discriminations after intensive practice. The behavioural aspects of such perceptual learning are well documented, especially in the visual system. Specificity for stimulus attributes clearly implicates an early cortical site, where receptive fields retain fine selectivity for these attributes; however, the neuronal correlates of a simple visual discrimination task remained unidentified. Here we report electrophysiological correlates in the primary visual cortex (V1) of monkeys for learning orientation identification. We link the behavioural improvement in this type of learning to an improved neuronal performance of trained compared to naive neurons. Improved long-term neuronal performance resulted from changes in the characteristics of orientation tuning of individual neurons. More particularly, the slope of the orientation tuning curve that was measured at the trained orientation increased only for the subgroup of trained neurons most likely to code the orientation identified by the monkey. No modifications of the tuning curve were observed for orientations for which the monkey had not been trained. Thus training induces a specific and efficient increase in neuronal sensitivity in V1.

Animals↗

PubMed2001

Inferior temporal neurons show greater sensitivity to nonaccidental than to metric shape differences.

It has long been known that macaque inferior temporal (IT) neurons tend to fire more strongly to some shapes than to others, and that different IT neurons can show markedly different shape preferences. Beyond the discovery that these preferences can be elicited by features of moderate complexity, no general principle of (nonface) object recognition had emerged by which this enormous variation in selectivity could be understood. Psychophysical, as well as computational work, suggests that one such principle is the difference between viewpoint-invariant, nonaccidental (NAP) and view-dependent, metric shape properties (MPs). We measured the responses of single IT neurons to objects differing in either a NAP (namely, a change in a geon) or an MP of a single part, shown at two orientations in depth. The cells were more sensitive to changes in NAPs than in MPs, even though the image variation (as assessed by wavelet-like measures) produced by the former were smaller than the latter. The magnitude of the response modulation from the rotation itself was, on average, similar to that produced by the NAP differences, although the image changes from the rotation were much greater than that produced by NAP differences. Multidimensional scaling of the neural responses indicated a NAP/MP dimension, independent of an orientation dimension. The present results thus demonstrate that a significant portion of the neural code of IT cells represents differences in NAPs rather than MPs. This code may enable immediate recognition of novel objects at new views.

Animals↗

PubMed2001

Encoding of categories by noncategory-specific neurons in the inferior temporal cortex.

In order to understand how the brain codes natural categories, e.g., trees and fish, recordings were made in the anterior part of the macaque inferior temporal (IT) cortex while the animal was performing a tree/nontree categorization task. Most single cells responded to exemplars of more than one category while other neurons responded only to a restricted set of exemplars of a given category. Since it is still not known which type of cells contribute and what is the nature of the code used for categorization in IT, we have performed an analysis on single-cell data. A Kohonen self-organizing map (SOM), which uses an unsupervised (competitive) learning algorithm, was used to study the single cell responses to tree and nontree images. Results from the Kohonen SOM indicated that the collected neuronal data consisting of spike counts was sufficient to account for a good level of categorization success (approximately 83%) when categorizing a group of 200 trees and nontrees. Contrary to intuition, the results of the investigation suggest that the population of category-specific neurons (neurons that respond only to trees or only to nontrees) was unimportant to the categorization. Instead, a large majority of the neurons that were most important to the categorization was found to belong to a class of more broadly tuned cells, namely, cells that responded to both categories but that favored one category over the other by seven or more images. A simple algebraic operation (without the Kohonen SOM) between the above-mentioned noncategory-specific neurons confirmed the contribution of these neurons to categorization. Thus, the modeling results suggest (1) that broadly tuned neurons are critical for categorization, and (2) that only one additional layer of processing is required to extract the categories from a population of IT neurons.

Animals↗

PubMed2001

The influence of synovial fluid on adenovirus-mediated gene transfer to the synovial tissue.

OBJECTIVE: To determine the effect of synovial fluid (SF) from rheumatoid arthritis (RA) patients on adenovirus type 5 (Ad5)-mediated gene transfer to synoviocytes, and to explore new strategies for vector development based on the neutralization data obtained. METHODS: SF was derived from 63 randomly selected R4 patients. Ten samples were used to study the effect of SF on Ad5-mediated gene transfer in synoviocytes. IgG and <100-kd fractions were purified from these 10 SF, and their effect on gene transfer was determined. Neutralizing activity against wild-type Ad5 (wt-Ad5), wt-Ad26, wt-Ad34, wt-Ad35, and wt-Ad48 was tested in the SF from the remaining 53 patients. RESULTS: Seven of 10 SF samples inhibited Ad5-mediated gene transfer. Purified antibodies exhibited inhibition patterns similar to those seen with unfractionated SF. In 5 of 10 SF samples, low molecular weight fractions inhibited gene transfer at low dilutions. Neutralization of wt-Ad35 by SF from RA patients was less frequent than neutralization of other wt-Ad tested (4% versus 42-72%; n = 53). CONCLUSION: SF from 70% of the RA patients contained neutralizing antibodies that hamper Ad5-mediated gene transfer to synoviocytes. The activity of neutralizing antibodies may be circumvented in the majority of RA patients when vectors based on an Ad35 backbone are used.

Adenoviridae↗

PubMed2001

Can neuroimaging really tell us what the human brain is doing? The relevance of indirect measures of population activity.

Neuroimaging studies using positron emission tomography (PET) and functional magnetic resonance imaging (fMRI) give an indication towards the localization of mental representations and processes in the human brain. It is not clear to what extent such global measures of neuronal activity, pooling across large populations of neurons, can reveal how certain computations are implemented by the neurons in such population ('computational neuroimaging'). Population activity is related tightly to single-cell activity when all neurons in the population have similar response properties. We describe some evidence from single-cell recordings in monkeys that indicates that neurons with similar response properties are not scattered randomly throughout the visual cortex. Notwithstanding this clustering, populations of nearby neurons are still rather heterogeneous, requiring some prudence in deriving single-cell response properties from population activity. The following review of recent neuroimaging studies of the visual system describes to what degree inferences about computations and representations can be drawn from these studies.

Animals↗

PubMed2001

Inferotemporal neurons represent low-dimensional configurations of parameterized shapes.

Behavioral studies with parameterized shapes have shown that the similarities among these complex stimuli can be represented using a low number of dimensions. Using psychophysical measurements and single-cell recordings in macaque inferotemporal (IT) cortex, we found an agreement between low-dimensional parametric configurations of shapes and the representation of shape similarity at the behavioral and neuronal level. The shape configurations, computed from both the perceived and neuron-based similarities, revealed a low number of dimensions and contained the same stimulus order as the parametric configurations. However, at a metric level, the behavioral and neural representations deviated consistently from the parametric configurations. These findings suggest an ordinally faithful but metrically biased representation of shape similarity in IT.

Action Potentials↗

PubMed2001

Improved adenovirus vectors for infection of cardiovascular tissues.

To identify improved adenovirus vectors for cardiovascular gene therapy, a library of adenovirus vectors based on adenovirus serotype 5 (Ad5) but carrying fiber molecules of other human serotypes, was generated. This library was tested for efficiency of infection of human primary vascular endothelial cells (ECs) and smooth muscle cells (SMCs). Based on luciferase, LacZ, or green fluorescent protein (GFP) marker gene expression, several fiber chimeric vectors were identified that displayed improved infection of these cell types. One of the viruses that performed particularly well is an Ad5 carrying the fiber of Ad16 (Ad5.Fib16), a subgroup B virus. This virus showed, on average, 8- and 64-fold-increased luciferase activities on umbilical vein ECs and SMCs, respectively, compared to the parent vector. GFP and lacZ markers showed that approximately 3-fold (ECs) and 10-fold (SMCs) more cells were transduced. Experiments performed with both cultured SMCs and organ cultures derived from different vascular origins (saphenous vein, iliac artery, left interior mammary artery, and aorta) and from different species demonstrated that Ad5.Fib16 consistently displays improved infection in primates (humans and rhesus monkeys). SMCs of the same vessels of rodents and pigs were less infectable with Ad5.Fib16 than with Ad5. This suggests that either the receptor for human Ad16 is not conserved between different species or that differences in the expression levels of the putative receptor exist. In conclusion, our results show that an Ad5-based virus carrying the fiber of Ad16 is a potent vector for the transduction of primate cardiovascular cells and tissues.

Adenoviruses, Human↗

PubMed2000

Spatial sensitivity of macaque inferior temporal neurons.

Recent findings in dorsal visual stream areas and computational work raise the question whether neurons at the end station of the ventral visual stream can code for stimulus position. The authors provide the first detailed, quantitative data on the spatial sensitivity of neurons in the anterior part of the inferior temporal cortex (area TE) in awake, fixating monkeys. They observed a large variation in receptive field (RF) size (ranging from 2.8 degrees to 26 degrees ). TE neurons differed in their optimal position, with a bias toward the foveal position. Moreover, the RF profiles of most TE neurons could be fitted well with a two-dimensional Gaussian function. Most neurons had only one region of high sensitivity and showed a smooth decline in sensitivity toward more distal positions. In addition, the authors investigated some of the possible determinants of such spatial sensitivity. First, testing with low-pass filtered versions of the stimuli revealed that the general preference for the foveal position and the size of the RFs was not due simply to TE neurons receiving input with a lower spatial resolution at more eccentric positions. The foveal position was still preferred after intense low-pass filtering. Second, although an increase in stimulus size consistently broadened spatial sensitivity profiles, it did not change the qualitative features of these profiles. Moreover, size selectivity of TE neurons was generally position invariant. Overall, the results suggest that TE neurons can code for the position of stimuli in the central region of the visual field.

Animals↗

PubMed2000

Selectivity for 3D shape that reveals distinct areas within macaque inferior temporal cortex.

The anterior part of the macaque inferior temporal cortex, area TE, occupies a large portion of the temporal lobe and is critical for object recognition. Thus far, no relation between anatomical subdivisions of TE and neuronal selectivity has been described. Here, we present evidence that neurons selective for three-dimensional (3D) shape are concentrated in the lower bank of the superior temporal sulcus, whereas neurons in lateral TE are generally unselective for 3D shape, though equally selective for 2D shape. These findings reveal that TE consists of at least two distinct areas, one of which processes a specific object property.

Animals↗

PubMed2000

Impairment in motion discrimination tasks is unrelated to amount of damage to superior temporal sulcus motion areas.

The behavioral role of the middle temporal (MT/V5) area and its satellites in motion processing is still unclear, particularly the degree to which MT/V5 proper is critical for different types of motion processing. Therefore, effects of small and large lesions in the caudal part of the superior temporal sulcus of macaque monkeys were compared for two tasks requiring different types of motion processing: a direction and a kinetic orientation discrimination. The small lesion was restricted to the peripheral representation of MT/V5 but included V4t, whereas the large lesion included all of MT/V5 and the medial superior temporal (MST) area as well as substantial parts of the floor of the superior temporal (FST) area. Both lesions resulted in significant and long-lasting impairment of direction discrimination but had a lesser effect on kinetic orientation discrimination. Thus the effects of small STS lesions on motion perception are much stronger than expected.

Animals↗

PubMed2000

Three-dimensional shape coding in inferior temporal cortex.

Neurons in the rostral lower bank of the superior temporal sulcus (TEs), part of the inferior temporal cortex, respond selectively to three-dimensional (3D) shapes. We have investigated how these neurons represent disparity-defined 3D structure. Most neurons were selective for either first-order (disparity gradients) or second-order (disparity curvature) disparities. The latter selectivity proved remarkably vulnerable to disparity discontinuities, such as sharp edges or steps in disparity. The majority of the neurons remained selective for small disparity variations within the stimulus. 3D shape selectivity was preserved when the frontoparallel position or the stimulus size was altered. Thus, in TEs, 3D shape is coded by first- and second-order disparity-selective neurons, which are highly sensitive to spatial variations of disparity.

Action Potentials↗

PubMed1999

Macaque inferior temporal neurons are selective for disparity-defined three-dimensional shapes.

Real-world objects are three-dimensional (3D). Yet, it is unknown whether the neurons of the inferior temporal cortex, which is critical for object recognition, are selective for the 3D shape of objects. We tested for such selectivity by comparing responses to stereo-defined curved 3D shapes derived from identical pairs of monocular images. More than one-third of macaque inferior temporal neurons were selective for 3D shape. In the vast majority of those neurons, this selectivity depended on the global binocular disparity gradient and not on the local disparity. Thus, inferior temporal cortex processes not only two-dimensional but also 3D shape information.

Animals↗

PubMed1999

Effect of image scrambling on inferior temporal cortical responses.

Imaging studies compared activations elicited by images of objects and scrambled versions of these same images in order to localize human brain regions involved in object recognition. Given these studies and the known role of macaque temporal cortex in object recognition, I determined the effect of image scrambling on the responses of macaque inferior temporal neurons. Images of natural objects were scrambled to different degrees. The response of most neurons decreased with increasing degrees of scrambling. In 26% of the neurons, response was reduced by 50% when the image was scrambled using only four parts, but most neurons tolerated higher degrees of scrambling, suggesting that these neurons code object parts instead of entire objects. These results are related to human functional imaging studies.

Action Potentials↗

PubMed1999

Temporal resolution deficits in the visual fields of MS patients.

We assessed the relationship between temporal resolution and MS-induced neuropathy. A diagnostic strategy comprising assessments of temporal resolution at 16 points in the extra-foveal visual field up to 12 degrees from the fovea was first compared with foveal temporal resolution and with a standard VEP procedure in the same MS patients. At the group level, foveal temporal resolution was less sensitive to demyelination than the 16-point diagnostic strategy, the detection rate of which was comparable to that of the VEP procedure. Cross-sensitivity of the VEP and the 16-point diagnostic procedure was low. Subsequently, the average severity of MS-induced temporal resolution deficits was studied at three retinal loci of the same size but different eccentricities. Foveal deficits were not significantly greater than more peripheral deficits within the central 12 degrees.

Adult↗

PubMed1999

Ganciclovir-mediated in vivo elimination of myeloid leukemic cells expressing the HSVtk gene induces HSVtk loss variants.

The in vivo elimination of suicide gene-expressing tumor cells with prodrug treatment can induce protective immunity against wild-type tumors. In this study, we determined the efficacy and safety of the in vivo elimination of HSVtk expressing cells with ganciclovir treatment of a bystander cell killing-insensitive leukemic cell line. The retroviral construct pLTk+NeoDeltaMo, containing the HSVtk gene and the NeoR gene in a bicistronic unit, was introduced into rat leukemic LT12 cells. LT12/Tk+N cells showed a 1000- to 10 000-fold increased sensitivity to ganciclovir in vitro. In vitro mixing experiments demonstrated that LT12 cells were not susceptible to bystander cell lysis by LT12/Tk+N-2 cells exposed to ganciclovir. Rats injected s.c. with cloned LT12/Tk+N-2 cells developed tumors reaching a diameter of 3-4 cm after 40 days. Rats treated with gan- ciclovir twice daily for 5 consecutive days starting at day 7 did not develop s.c. tumors. Large established s.c. LT12/Tk+N-2 tumors completely regressed after ganciclovir treatment. However, recurrences of s.c. tumors were observed that were no longer sensitive to ganciclovir treatment. In vitro analysis of aspirates from the recurrent tumors demonstrated loss of HSVtk expression. In vitro culture of LT12/Tk+N-2 cells in soft agar in the presence of ganciclovir indicated that the frequency with which HSVtk-loss variants occurred is approximately one per 104 cells. The in vivo occurrence of HSVtk-loss variants escaping ganciclovir-induced elimination may have important implications for vaccination protocols using HSVtk gene expressing tumor cells that are not susceptible to bystander cell killing.

Animals↗