MHC class I-peptide tetramers are currently utilised to characterize CD8+ T cell responses at single cell level. The generation and use of MHC class II tetramers to study antigen-specific CD4+ T ...cells appears less straightforward. Most MHC class II tetramers are produced with a homogeneously built-in peptide, reducing greatly their flexibility of use. We attempted the generation of "empty" functional HLA-DR*1101 tetramers, receptive for loading with synthetic peptides by incubation. No such reagent is in fact available for this HLA-DR allele, one of the most frequent in the Caucasian population.
We compared soluble MHC class II-immunoglobulin fusion proteins (HLA-DR*1101-Ig) with soluble MHC class II protein fused with an optimised Bir site for enzymatic biotynilation (HLA-DR*1101-Bir), both produced in insect cells. The molecules were multimerised by binding fluorochrome-protein A or fluorochrome-streptavidin, respectively. We find that HLA-DR*1101-Bir molecules are superior to the HLA-DR*1101-Ig ones both in biochemical and functional terms. HLA-DR*1101-Bir molecules can be pulsed with at least three different promiscuous peptide epitopes, derived from Tetanus Toxoid, influenza HA and the tumour associated antigen MAGE-3 respectively, to stain specific CD4+ T cells. Both staining temperature and activation state of CD4+ T cells are critical for the binding of peptide-pulsed HLA-DR*1101-Bir to the cognate TCR.
It is therefore possible to generate a soluble recombinant HLA-DR*1101 backbone that is receptive for loading with different peptides to stain specific CD4+ T cells. As shown for other HLA-DR alleles, we confirm that not all the strategies to produce soluble HLA-DR*1101 multimers are equivalent.
Maspin, a member of the serpin family of protease inhibitors, is involved in key processes of cancer progression. Its biological activity seems to be cancer and compartment specific, with the protein ...acting either as a suppressor or as a tumor promoter in different cancer types. Characterization of maspin expression and its sub-cellular localization in melanoma is missing, hence, we aim to investigate its possible association with melanoma prognostic factors and disease progression. Nuclear and cytoplasmic maspin expression were evaluated on 60 nevi, 152 primary lesions, and 106 melanoma metastases using tissue microarrays and immunohistochemistry. The association between maspin immunoreactivity and patient's clinic-pathological features was evaluated. Multivariate logistic models and survival analyses were performed for maspin expression in primary melanomas. Nuclear maspin was detected in 8% nevi, 49% primary melanomas, and 28% metastases, whereas cytoplasmic maspin in 12% nevi, 18% primary lesions, and 9% metastases. In univariate analysis, nuclear maspin expression in primary melanomas was significantly associated with melanoma prognostic factors (nodular histotype, tumor thickness, mitotic rate, and ulceration) and disease stage, whereas cytoplasmic maspin was observed at higher frequency in thin superficial spreading melanomas, without mitosis. In multivariate analysis, nuclear maspin remained significantly associated with risk of developing a tumor prone to disease progression and, accordingly, with significantly shorter disease-free and overall survival. In this study, maspin was expressed at highest frequency in primary lesions and when expressed in the nuclei, was significantly associated with poor prognostic markers, melanoma recurrence, and worse survival. The present study suggests a tumor-suppressive effect of cytoplasmic maspin and a tumor-promoting effect of nuclear maspin, which open the discussion on its potential use in cancer therapy.
Secretory immunoglobulin A (SIgA) represents a first line of defense against mucosal pathogens by limiting their entrance. By using different strains of Salmonella typhimurium that target the two ...mechanisms of bacterial entry (microfold cell M cell- or dendritic cell-mediated), we demonstrated here that the distribution of bacteria after oral infection directed the type of induced immune response. M cell-penetrating invasive, but not noninvasive, S. typhimurium was found in large numbers in Peyer's patches (PPs), leading to the activation of immune cells and the release of fecal IgA. In contrast, both strains of bacteria were equally capable of reaching the mesenteric lymph node and the spleen and inducing IgG responses. These data suggest that PPs are absolutely required for the initiation of an IgA response to Salmonella, whereas they are dispensable for a systemic response. This compartmentalization could allow the fast generation of both mucosal and systemic acquired immunity to pathogens.
Within the scope of the ongoing efforts to fight climate change, the application of multi-robot systems to environmental mapping and monitoring missions is a prominent approach aimed at increasing ...exploration efficiency. However, the application of such systems to gas sensing missions has yet to be extensively explored and presents some unique challenges, mainly due to the hard-to-sense and expensive-to-model nature of gas dispersion. For this paper, we explored the application of a multi-robot system composed of rotary-winged nano aerial vehicles to a gas sensing mission. We qualitatively and quantitatively analyzed the interference between different robots and the effect on their sensing performance. We then assessed this effect, by deploying several algorithms for 3D gas sensing with increasing levels of coordination in a state-of-the-art wind tunnel facility. The results show that multi-robot gas sensing missions can be robust against documented interference and degradation in their sensing performance. We additionally highlight the competitiveness of multi-robot strategies in gas source location performance with tight mission time constraints.
The ability to sense airborne pollutants with mobile robots provides a valuable asset for domains such as industrial safety and environmental monitoring. Oftentimes, this involves detecting how ...certain gases are spread out in the environment, commonly referred to as a gas distribution map, to subsequently take actions that depend on the collected information. Since the majority of gas transducers require physical contact with the analyte to sense it, the generation of such a map usually involves slow and laborious data collection from all key locations. In this regard, this paper proposes an efficient exploration algorithm for 2D gas distribution mapping with an autonomous mobile robot. Our proposal combines a Gaussian Markov random field estimator based on gas and wind flow measurements, devised for very sparse sample sizes and indoor environments, with a partially observable Markov decision process to close the robot's control loop. The advantage of this approach is that the gas map is not only continuously updated, but can also be leveraged to choose the next location based on how much information it provides. The exploration consequently adapts to how the gas is distributed during run time, leading to an efficient sampling path and, in turn, a complete gas map with a relatively low number of measurements. Furthermore, it also accounts for wind currents in the environment, which improves the reliability of the final gas map even in the presence of obstacles or when the gas distribution diverges from an ideal gas plume. Finally, we report various simulation experiments to evaluate our proposal against a computer-generated fluid dynamics ground truth, as well as physical experiments in a wind tunnel.
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