Novel CCL21-Vault Nanocapsule Intratumoral Delivery Inhibits Lung Cancer Growth

Based on our preclinical findings, we are assessing the efficacy of intratumoral injection of dendritic cells (DC) transduced with an adenoviral vector expressing the secondary lymphoid chemokine (CCL21) gene (Ad-CCL21-DC) in a phase I trial in advanced non-small cell lung cancer (NSCLC). While this approach shows immune enhancement, the preparation of autologous DC for CCL21 genetic modification is cumbersome, expensive and time consuming. We are evaluating a non-DC based approach which utilizes vault nanoparticles for intratumoral CCL21 delivery to mediate antitumor activity in lung cancer.Here we describe that vault nanocapsule platform for CCL21 delivery elicits antitumor activity with inhibition of lung cancer growth. Vault nanocapsule packaged CCL21 (CCL21-vaults) demonstrated functional activity in chemotactic and antigen presenting activity assays. Recombinant vaults impacted chemotactic migration of T cells and this effect was predominantly CCL21 dependent as CCL21 neutralization abrogated the CCL21 mediated enhancement in chemotaxis. Intratumoral administration of CCL21-vaults in mice bearing lung cancer enhanced leukocytic infiltrates (CXCR3(+)T, CCR7(+)T, IFNγ(+)T lymphocytes, DEC205(+) DC), inhibited lung cancer tumor growth and reduced the frequencies of immune suppressive cells [myeloid derived suppressor cells (MDSC), T regulatory cells (Treg), IL-10 T cells]. CCL21-vaults induced systemic antitumor responses by augmenting splenic T cell lytic activity against parental tumor cells.This study demonstrates that the vault nanocapsule can efficiently deliver CCL21 to sustain antitumor activity and inhibit lung cancer growth. The vault nanocapsule can serve as an "off the shelf" approach to deliver antitumor cytokines to treat a broad range of malignancies

Non-linear behaviour of concrete beams reinforced with GFRP and CFRP bars grouted in sleeves

The low-quality bond between fibre reinforced polymer (FRP) bars and surrounding concrete has drawn the attention of many researchers. The use of high-strength materials such as the grout in the intersection of FRP bars and surrounding concrete can effectively prevent any slippage once they are in contact and subsequently increase the bond quality. Therefore, this study was numerically focused on the flexural behaviour of concrete beams reinforced with glass fibre reinforced polymer (GFRP) and carbon fibre reinforced polymer (CFRP) bars, grouted only in the pure bending zone and along the whole beam length. The numerical outputs revealed that the grouted GFRP bars propagated the maximum principal stress in high-strength concrete beams, but not as much as that in normal-strength concrete specimens. In addition, the stress distribution in the grout, created only in the pure bending zone, was nearly constant at the ultimate moment. For the grout, developed along the whole beam length, this stress increased by approaching the mid-span of the concrete beam. Furthermore, at the ultimate moment, the tensile stress of 12-mm diameter CFRP bars was about 3.5 times more than that of the 16-mm diameter CFRP bars, leading to the generation of difference between failure modes of concrete specimens reinforced with various diameters of CFRP bars

Personalization in Skipforward, an Ontology-Based Distributed Annotation System

Abstract. Skipforward is a distributed annotation system allowing users to enter and browse statements about items and their features. Items can be things such as movies or books; item features are the genre of a movie or the storytelling pace of a book. Whenever multiple users annotate the same item with a statement about the same feature, these individual statements get aggregated by the system. For aggregation, individual user statements are weighted according to a competence metric based on the constrained Pearson correlation, adapted for Skipforward data: A user gets assigned high competence with regard to the feature in question if, for other items and the same feature type, he had a similar opinion to the current user. Since the competence metric is dependent on the user currently viewing the data, the user’s view of the data is completely personalized. In this paper, the personalization aspect as well as the item and expert recommender are presented.

Genes differenially expressed in VK2 cells treated with proinflammatory/immunomodulatory compounds.

<p>Genes differenially expressed in VK2 cells treated with proinflammatory/immunomodulatory compounds.</p

Functional categories of the PIC/NIC discriminatory genes^a.

<p>^<b>a</b>Classification is based on IPA functional analysis and published literature. P values are estimated by IPA</p><p>Functional categories of the PIC/NIC discriminatory genes<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0128557#t004fn001" target="_blank">^a</a>.</p

Real-time qPCR validation of changes in expression of eight selected genes observed in microarray analysis.

<p>Bars represent the mean ± SD of fold change relative to growth medium control. At least 3 independent experiments were performed for each treatment. All genes were normalized to GAPDH. Asterisks placed vertically denote p values for each PIC treatment relative to HEC used as a reference. (***p<0.0005, **p<0.005, *p< 0.05; Student t-test)</p

Top network of Vk2 PIC/NIC discriminatory genes generated by Ingenuity Pathway Analysis.

<p>Red/pink color indicates upregulation of the genes (microarray data). Connections of NFkB complex with other genes is shown in blue color.</p

Transcription profile of the 20 discriminatory genes expression in Vk2 cells exposed to candidate microbicides and selected PICs and NICs.

<p>Columns represent treatments, rows represent genes. Gene expression levels are indicated by color: red is for upregulation and green is for downregulation. Expression data are averages from at least six experiments/microarrays for each treatment. Clustering based on 20 PIC/NIC discriminatory genes places C31G (known as causing inflammatory response) to the PIC category, while dextran sulfate (DS) and cellulose sulfate (CS)—into the NIC group.</p

Diagrams showing the number of significantly altered probesets indentified by microarray gene profiling of Vk2 cells exposed to PIC and NIC.

<p>Total number of the altered probeserts for each treatment/category is shown in brackets (a gene can be represented by more than one probeset).</p