Potential of Ni(II)-NTA-modified poly(ethylene imine) glycopolymers as carrier system for future dendritic cell-based immunotherapy.

Dendritic cells (DCs) play a crucial role in the development of cell-mediated immunotherapy due to their ability to induce and maintain strong immune responses. In our study, we evaluated a biocompatible Ni(II)-NTA-modified poly(ethylene imine) dendritic glycopolymer (Ni(II)-NTA-DG) as new carrier system to increase the antigen uptake into iDCs for future DC-based immunotherapy. Ni(II)-NTA-DG led to an increase in His6-Gp160 uptake in monocytes and iDCs, where His6-Gp160 is localized in the early endosomal and lysosomal compartments. Ni(II)-NTA-DG and the formed polyplexes induced an activation of iDCs, showing an increasing expression of costimulatory molecules CD86, CD80, and proinflammatory cytokines IL-6 and IL-8. Beside no influencing effect of Ni(II)-NTA-DG and polyplexes on the maturation of antigen-bearing DCs, the mature peptide bearing DCs remained their ability to migrate along a gradient of CCR7 ligands. Thus, Ni(II)-NTA-DG with advancing biological properties is a promising carrier system for the future application in DC-based immunotherapy.

IDO1 and IDO2 gene expression analysis by quantitative polymerase chain reaction.

Immunomodulatory properties of IDO1 relate to tryptophan catabolism. The degradation of tryptophan by IDO1 leads to suppression of T cell responses. Recently, another enzyme with IDO-like activity, indoleamine 2,3-dioxygenase-like-protein 1 (INDOL1, IDO2), has been described in both mice and humans. In order to study the gene expression of IDO1 and IDO2, we have developed a quantitative PCR (qPCR) assay. In an exploratory application to the study of the differential expression of IDO1 and IDO2 by professional antigen-presenting cells and MSCs (mesenchymal stromal cells) under the influence of interferon-γ (IFN-γ) and T-lymphocyte conditioned media (TCM), substantial differences were observed. IDO expression measured by qPCR was valid and reliable in the cell types investigated. Further studies are needed to delineate factors driving IDO expression in MSCs.

Bortezomib significantly impairs the immunostimulatory capacity of human myeloid blood dendritic cells.

Bortezomib is a potent drug for the treatment of multiple myeloma. Its anti-tumor activity is mediated by proteasome inhibition leading to decreased cell proliferation and induction of apoptosis. However, an unimpaired proteasomal function plays a crucial role for the induction of anti-tumor immunity by dendritic cells (DCs), which are currently used for therapeutic vaccination against various tumors including myeloma. In the present study, we investigated the impact of bortezomib on the immunostimulatory capacity of 6-sulfo LacNAc (slan) DCs, which represent a major subset of human blood DCs. We demonstrated that this proteasome inhibitor efficiently impairs the spontaneous in vitro maturation of slanDCs and the release of tumor necrosis factor (TNF)-alpha as well as interleukin (IL)-12 upon lipopolysaccharide (LPS) stimulation. Functional data revealed that bortezomib profoundly inhibits slanDC-induced proliferation and differentiation of CD4(+) T cells. In addition, the capacity of slanDCs to promote interferon-gamma secretion and tumor-directed cytotoxicity of natural killer (NK) cells is markedly impaired by bortezomib. These results provide evidence that bortezomib significantly reduces the ability of native human blood DCs to regulate innate and adaptive anti-tumor immunity and may have implications for the design of therapeutic strategies combining DC vaccination and bortezomib treatment.

Identification of SOX2 as a novel glioma-associated antigen and potential target for T cell-based immunotherapy.

Prognosis for patients suffering from malignant glioma has not substantially improved. Specific immunotherapy as a novel treatment concept critically depends on target antigens, which are highly overexpressed in the majority of gliomas, but the number of such antigens is still very limited. SOX2 was identified by screening an expression database for transcripts that are overexpressed in malignant glioma, but display minimal expression in normal tissues. Expression of SOX2 mRNA was further investigated in tumour and normal tissues by real-time PCR. Compared to cDNA from pooled normal brain, SOX2 was overexpressed in almost all (9 out of 10) malignant glioma samples, whereas expression in other, non-malignant tissues was almost negligible. SOX2 protein expression in glioma cell lines and tumour tissues was verified by Western blot and immunofluorescence. Immunohistochemistry demonstrated SOX2 protein expression in all malignant glioma tissues investigated ranging from 6 to 66% stained tumour cells. Human leucocyte antigen-A(*)0201-restricted SOX2-derived peptides were tested for the activation of glioma-reactive CD8+ cytotoxic T lymphocytes (CTLs). Specific CTLs were raised against the peptide TLMKKDKYTL and were capable of lysing glioma cells. The abundant and glioma-restricted overexpression of SOX2 and the generation of SOX2-specific and tumour-reactive CTLs may recommend this antigen as target for T-cell-based immunotherapy of glioma.

Integrating two-dimensional paths: do desert ants process distance information in the absence of celestial compass cues?

When performing foraging trips desert ants of the genus Cataglyphis continuously process and update a ;home vector' that enables them to return to their nest on the shortest route. This capacity of path integration requires two types of information: (i) information about the travelling directions, and (ii) odometric information about the distances travelled in a particular direction. We have investigated how these two necessary pieces of information interact within the path integration processor. The specific question is: how do the ants process distance information if there is no simultaneous input from the sky compass available. Ants were trained to forage in a ;Z'-shaped channel system, the three segments of which joined at right angles. Individual animals were transferred from the feeder to a test field where their homing paths could be observed. In the crucial tests the middle segment of the maze was covered by orange Perspex that did not transmit the UV part of the spectrum, and thus precluded the perception of polarization patterns. Changes of the ant's processing of odometric information within this channel segment directly translate into a change in homing direction on the test field. The results indicate that the odometric information about travelling distance is largely ignored for path integration if there is no simultaneous input from the sky-view-based compass. They further show that idiothetic information cannot adequately substitute for the polarization compass to infer travelling directions.

Look and turn: landmark-based goal navigation in honey bees.

This report describes the piloting mechanisms employed by honey bees during their final approach to a goal. Conceptually applying a bottom-up approach, we systematically varied the position, number and appearance landmarks associated with a rewarded target location within a large, homogenous flight tent. The flight behavior measured under various conditions is well explained with visuo-motor control loops that link perceived landmarks with appropriate turning responses. This view is consistent with the requirement of prolonged reinforcement learning for efficient goal navigation. A simple model is able to provide a comprehensive explanation for diverse flight patterns that range from convoluted searching behavior to highly idiosyncratic approaches, depending on the experimental context. Our results challenge the prevalent notion that honey bees employ image matching for visual guidance toward a goal site. Basic visuo-motor control loops may better meet the high demands for robust and fast flight control, which could serve as a powerful bio-mimetic design principle for micro-robotic aircraft.

The ant's estimation of distance travelled: experiments with desert ants, Cataglyphis fortis.

Foraging desert ants, Cataglyphis fortis, monitor their position relative to the nest by path integration. They continually update the direction and distance to the nest by employing a celestial compass and an odometer. In the present account we addressed the question of how the precision of the ant's estimate of its homing distance depends on the distance travelled. We trained ants to forage at different distances in linear channels comprising a nest entrance and a feeder. For testing we caught ants at the feeder and released them in a parallel channel. The results show that ants tend to underestimate their distances travelled. This underestimation is the more pronounced, the larger the foraging distance gets. The quantitative relationship between training distance and the ant's estimate of this distance can be described by a logarithmic and an exponential model. The ant's odometric undershooting could be adaptive during natural foraging trips insofar as it leads the homing ant to concentrate the major part of its nest-search behaviour on the base of its individual foraging sector, i.e. on its familiar landmark corridor.

Long-distance navigation in the wandering desert spider Leucorchestris arenicola: can the slope of the dune surface provide a compass cue?

Males of the nocturnal spider Leucorchestris arenicola (Araneae: Sparassidae) wander long distances over seemingly featureless dune surfaces in the Namib Desert searching for females. The spiders live in burrows to which they return after nearly every such excursion. While the outward path of an excursion may be a meandering search, the return path is often a nearly straight line leading towards the burrow. This navigational behaviour resembles that of path integration known from other arthropods, though on a much larger scale (over tens to hundreds of meters). Theoretically, precise navigation by path integration over long distances requires an external compass in order to adjust for inevitable accumulation of navigational errors. As a first step towards identifying any nocturnal compass cues used by the male spiders, a method for detailed 3-D recordings of the spider's paths was developed. The 3-D reconstructions of the paths revealed details about the processes involved in the spiders' nocturnal way of navigation. Analyses of the reconstructed paths suggest that gravity (slope of the dune surface) is an unlikely parameter used in path integration by the L. arenicola spiders.

Desert ant navigation: how miniature brains solve complex tasks.

This essay presents and discusses the state of the art in studies of desert ant (Cataglyphis) navigation. In dealing with behavioural performances, neural mechanisms, and ecological functions these studies ultimately aim at an evolutionary understanding of the insect's navigational toolkit: its skylight (polarization) compass, its path integrator, its view-dependent ways of recognizing places and following landmark routes, and its strategies of flexibly interlinking these modes of navigation to generate amazingly rich behavioural outputs. The general message is that Cataglyphis uses path integration as an egocentric guideline to acquire continually updated spatial information about places and routes. Hence, it relies on procedural knowledge, and largely context-dependent retrieval of such knowledge, rather than on all-embracing geocentred representations of space.

Sympatry and allopatry in two desert ant sister species: how do Cataglyphis bicolor and C. savignyi coexist?

Two extremely morphologically similar sister species of desert ants, Cataglyphis bicolor and C. savignyi, exhibit broadly overlapping distributional ranges within Tunisia. In order to analyse the microhabitats of C. bicolor and C. savignyi within the sympatric and allopatric areas of both ant species, the plant species located at 113 different nest sites of the two ant species were determined. In the sympatric area, the two species exhibit a clear-cut nest site segregation. This is not the case in the allopatric areas. Hence the two species differentiate their microhabitat only when they are sympatric. The plant species associated mainly with the nest sites of C. bicolor indicate that this species prefers a type of vegetation that needs irrigation. This is in contrast to the nest sites of C. savignyi, which are usually found around plants that characterize typical dry steppe areas. As the ants' foraging paths recorded in the sympatric area reveal, C. bicolor performs significantly shorter foraging runs with respect to both length and time, and covers a much smaller foraging range than C. savignyi does. This result reflects the fact that the microhabitat occupied by the colonies of C. bicolor is richer in food abundance. When direct interspecific interactions were investigated by placing a bait midway between two heterospecific nests, C. bicolor foragers dominated over those of C. savignyi. The same dominance of C. bicolor over C. savignyi occurred in laboratory experiments. These results suggest that the dominant species drives the subordinate one out of the high quality microhabitats, and that the subordinate species is forced to survive in the less lucrative habitats. In conclusion, coexistence seems to be maintained by the asymmetric competitive relationship between the two species and the fact that the subordinate species has the ability to endure in the less favourable microhabitat.

Local vectors in desert ants: context-dependent landmark learning during outbound and homebound runs.

Desert ants, Cataglyphis fortis, associate nestward-directed vector memories (local vectors) with the sight of landmarks along a familiar route. This view-based navigational strategy works in parallel to the self-centred path integration system. In the present study we ask at what temporal stage during a foraging journey does the ant acquire nestward-directed local vector information from feeder-associated landmarks: during its outbound run to a feeding site or during its homebound run to the nest. Tests performed after two reversed-image training paradigms revealed that the ants associated such vectors exclusively with landmarks present during their homebound runs.

Do familiar landmarks reset the global path integration system of desert ants?

It is often suggested that animals may link landmark memories to a global coordinate system provided by path integration, thereby obtaining a map-like representation of familiar terrain. In an attempt to discover if desert ants form such associations we have performed experiments that test whether desert ants recall a long-term memory of a global path integration vector on arriving at a familiar food site. Ants from three nests were trained along L-shaped routes to a feeder. Each route was entirely within open-topped channels that obscured all natural landmarks. Conspicuous artificial landmarks were attached to the channelling that formed the latter part of the route. The homeward vectors of ants accustomed to the route were tested with the foodward route, either as in training, or with the first leg of the L shortened or extended. These ants were taken from the feeder to a test area and released, whereupon they performed a home vector. If travelling the latter part of a familiar route and arriving at a familiar food site triggers the recall of an accustomed home vector, then the home vector should be the same under both test conditions. We find instead that the home vector tended to reflect the immediately preceding outward journey. In conjunction with earlier work, these experiments led us to conclude in the case of desert ants that landmark memories do not prime the recall of long-term global path integration memories. On the other hand, landmark memories are known to be linked to local path integration vectors that guide ants along a segment of a route. Landmarks thus seem to provide procedural information telling ants what action to perform next but not the positional information that gives an ant its location relative to its nest.

Calibration processes in desert ant navigation: vector courses and systematic search.

This study investigates the ability of desert ants to adapt their path integration system to an "open-jaw" training paradigm, in which the point of arrival (from the nest) does not coincide with the point of departure (to the nest). Upon departure the ants first run off their home vector and then start a systematic search for the nest. Even if they are subjected to this training-around-a-circuit procedure for more than 50 times in succession, they never adopt straight homeward courses towards the nest. Their path integration vector gets slightly recalibrated (pointing a bit closer to the nest), and their search pattern gets asymmetric (with its search density peak shifted towards the nest), but the bipartite structure of the inbound trajectory invariably remains. These results suggest (1). that the ants cannot learn separate inbound and outbound vectors (i.e. vectors that are not 180 degrees reversals of each other), (2). that the recalibrated vector is dominated by the ant's outbound course, (3). that the recalibration of the vector and the modification of the search geometry are fast and flexible processes occurring whenever the ant experiences a mismatch between the stored and actual states of its path integrator.

Distance estimation in the third dimension in desert ants.

Desert ants of the genus Cataglyphis perform large-scale foraging excursions from which they return to their nest by path integration. They do so by integrating courses steered and the distances travelled into a continually updated home vector. While it is known that the angular orientation is based on skylight cues, it still is largely enigmatic how the ants measure distances travelled. We extended the ants' task into the third dimension by training them to walk within an array of uphill and downhill channels, and later testing them on flat terrain, or vice versa. In these tests the ants indicated homing distances that did not correspond to the distances actually travelled, but to the ground distances; that is, to the sum of the horizontal projections of the uphill and downhill segments of the ants' paths. These results suggest a much more sophisticated mechanism of distance estimation than hitherto thought. The ants must be able to measure the slopes of undulating terrain and to integrate this information into their "odometer" for the distance estimation process.

How the clear-sky angle of polarization pattern continues underneath clouds: full-sky measurements and implications for animal orientation.

One of the biologically most important parameters of the cloudy sky is the proportion P of the celestial polarization pattern available for use in animal navigation. We evaluated this parameter by measuring the polarization patterns of clear and cloudy skies using 180 degrees (full-sky) imaging polarimetry in the red (650 nm), green (550 nm) and blue (450 nm) ranges of the spectrum under clear and partly cloudy conditions. The resulting data were compared with the corresponding celestial polarization patterns calculated using the single-scattering Rayleigh model. We show convincingly that the pattern of the angle of polarization (e-vectors) in a clear sky continues underneath clouds if regions of the clouds and parts of the airspace between the clouds and the earth surface (being shady at the position of the observer) are directly lit by the sun. The scattering and polarization of direct sunlight on the cloud particles and in the air columns underneath the clouds result in the same e-vector pattern as that present in clear sky. This phenomenon can be exploited for animal navigation if the degree of polarization is higher than the perceptual threshold of the visual system, because the angle rather than the degree of polarization is the most important optical cue used in the polarization compass. Hence, the clouds reduce the extent of sky polarization pattern that is useful for animal orientation much less than has hitherto been assumed. We further demonstrate quantitatively that the shorter the wavelength, the greater the proportion of celestial polarization that can be used by animals under cloudy-sky conditions. As has already been suggested by others, this phenomenon may solve the ultraviolet paradox of polarization vision in insects such as hymenopterans and dipterans. The present study extends previous findings by using the technique of 180 degrees imaging polarimetry to measure and analyse celestial polarization patterns.

Polarization vision--a uniform sensory capacity?

In this concept paper, three scenarios are described in which animals make use of polarized light: the underwater world, the water surface and the terrestrial habitat vaulted by the pattern of polarized light in the sky. Within these various visual environments, polarized light is used in a number of ways that make quite different demands on the neural circuitries mediating these different types of behaviour. Apart from some common receptor and pre-processing mechanisms, the underlying neural mechanisms may differ accordingly. Often, information about chi (the angle of polarization), d (the degree of polarization) and lambda (the spectral content) might not --and need not--be disentangled. Hence, the hypothesis entertained in this account is that polarization vision comes in various guises, and that the answer to the question posed in the title is most probably no.