Nanoparticles for efficient uptake by human dendritic cells and T lymphocytes to be used in adoptive immunotherapy

Dendritic cells (DCs) are the most potent cell type for capture, processing, and presentation of antigens. They are able to activate naïve T cells as well as to initiate memory T-cell immune responses. T lymphocytes are key elements in eliciting cellular immunity against bacteria and viruses as well as in the generation of anti-tumor and anti-leukemia immune responses. Because of their central position in the immunological network, specific manipulations of these cell types provide promising possibilities for novel immunotherapies. Nanoparticles (NP) that have just recently been investigated for use as carriers of drugs or imaging agents, are well suited for therapeutic applications in vitro and also in vivo since they can be addressed to cells with a high target specificity upon surface functionalization. As a first prerequisite, an efficient in vitro labeling of cells with NP has to be established. In this work we developed protocols allowing an effective loading of human monocyte-derived DCs and primary antigen-specific T cells with newly designed NP without affecting biological cell functions. Polystyrene NP that have been synthesized by the miniemulsion technique contained perylenmonoimide (PMI) as a fluorochrome, allowing the rapid determination of intracellular uptake by flow cytometry. To confirm intracellular localization, NP-loaded cells were analyzed by confocal laser scanning microscopy (cLSM) and transmission electron microscopy (TEM). Functional analyses of NP-loaded cells were performed by IFN-γ ELISPOT, 51Chromium-release, and 3H-thymidine proliferation assays. In the first part of this study, we observed strong labeling of DCs with amino-functionalized NP. Even after 8 days 95% of DCs had retained nanoparticles with a median fluorescence intensity of 67% compared to day 1. NP loading did not influence expression of cell surface molecules that are specific for mature DCs (mDCs) nor did it influence the immunostimulatory capacity of mDCs. This procedure did also not impair the capability of DCs for uptake, processing and presentation of viral antigens that has not been shown before for NP in DCs. In the second part of this work, the protocol was adapted to the very different conditions with T lymphocytes. We used leukemia-, tumor-, and allo-human leukocyte antigen (HLA) reactive CD8+ or CD4+ T cells as model systems. Our data showed that amino-functionalized NP were taken up very efficiently also by T lymphocytes, which usually had a lower capacity for NP incorporation compared to other cell types. In contrast to DCs, T cells released 70-90% of incorporated NP during the first 24 h, which points to the need to escape from intracellular uptake pathways before export to the outside can occur. Preliminary data with biodegradable nanocapsules (NC) revealed that encapsulated cargo molecules could, in principle, escape from the endolysosomal compartment after loading into T lymphocytes. T cell function was not influenced by NP load at low to intermediate concentrations of 25 to 150 μg/mL. Overall, our data suggest that NP and NC are promising tools for the delivery of drugs, antigens, and other molecules into DCs and T lymphocytes.

Micropolar flow in a porous channel with high mass transfer

Two dimensional flow of a micropolar fluid in a porous channel is investigated. The flow is driven by suction or injection at the channel walls, and the micropolar model due to Eringen is used to describe the working fluid. An extension of Berman's similarity transform is used to reduce the governing equations to a set of non-linear coupled ordinary differential equations. The latter are solved for large mass transfer via a perturbation analysis where the inverse of the cross-flow Reynolds number is used as the perturbing parameter. Complementary numerical solutions for strong injection are also obtained using a quasilinearisation scheme, and good agreement is observed between the solutions obtained from the perturbation analysis and the computations.

The impact of unique characteristics of projects and project-based organisations on knowledge transfer

Knowledge has been recognised as an important organisational asset that increases in value when shared; the opposite to other organisational assets which decrease in value during their exploitation. Effective knowledge transfer in organisations helps to achieve and maintain competitive advantage and ultimately organisational success. So far, the research on knowledge transfer has focused on traditional (functional) organisations. Only recently has attention been directed towards knowledge transfer in projects. Existing research on project learning has recognised the need for knowledge transfer within and across projects in project-based organisations (PBOs). Most projects can provide valuable new knowledge from unexpected actions, approaches or problems experienced during the project phases. The aim of this paper is to demonstrate the impact of unique projects characteristics on knowledge transfer in PBO. This is accomplished through review of the literature and a series of interviews with senior project practitioners. The interviews complement the findings from the literature. Knowledge transfer in projects occurs by social communication and transfer of lessons learned where project management offices (PMOs) and project managers play significant roles in enhancing knowledge transfer and communication within the PBO and across projects. They act as connectors between projects and the PBO ‘hub’. Moreover, some project management processes naturally facilitate knowledge transfer across projects. On the other hand, PBOs face communication challenges due to unique and temporary characteristics of projects. The distance between projects and the lack or weakness of formal links across projects, create communication problems that impede knowledge transfer across projects. The main contribution of this paper is to demonstrate that both social communication and explicit informational channels play important role in inter-project knowledge transfer. Interviews also revealed the important role organisational culture play in knowledge transfer in PBOs.