Susanne Frank

Activated γδ T cells express the natural cytotoxicity receptor natural killer p44 and show cytotoxic activity against myeloma cells

γδ T cells account for up to 10% of T lymphocytes in the peripheral blood of healthy donors. They can be activated by cytokines such as interleukin (IL)‐2, IL‐12 and IL‐15, express natural killer (NK) cell markers such as NKG2D and show cytotoxic activity against several tumour cells, including multiple myeloma. Here, we present activated polyclonal γδ T cells from healthy donors with an NK T cell‐like phenotype expressing the natural cytotoxicity receptor NKp44. Natural cytotoxicity receptors NKp30, NKp44 and NKp46 have been regarded as specific NK receptors; only two γδ T cell clones described so far expressed NKp44. Isolated polyclonal γδ T cells cultured for 7 days according to the cytokine‐induced killer cell (CIK) protocol with additional IL‐15 revealed a surface expression of NKp44 of 8 ± 7% (n = 22). This could be confirmed by detection of NKp44 mRNA by reverse transcription–polymerase chain reaction (RT–PCR). γδ T cells exhibited a marked cytotoxic activity against myeloma cells, which could be reduced by inhibition of NKp44. To our knowledge, this is the first description of the expression of NKp44 on polyclonal γδ T cells.

Assessment of the effects of forest land use strategies on the provision of ecosystem services at regional scale.

This paper presents results of a case study in Middle Saxony, Germany, where the impact of conversion, afforestation and alternatively introduction of short rotation coppice areas on the provision of ecosystem services was tested in a spatially inexplicit and a spatially explicit way to formulate recommendations for regional planning. While the spatially inexplicit testing did not lead to clear results regarding to what degree forests or short rotation coppice areas are desirable and applicable, the spatially explicit testing revealed that an increase in the forest area or area with short rotation coppice by 29.7% in unstructured agriculturally dominated Loess regions, 14.4% in more topographically structured parts in the North-East of the model region and 23.6% in its mountainous parts would be beneficial. Potentially resulting losses in the provision of bioresources and regional economy can be considerably reduced by replacing afforestation areas with short rotation coppice. In summary, we found that the spatially explicit analysis of land use scenarios in combination with a more detailed land use classification and including an assessment of changes in land use pattern gave us an improved basis for assessing different possible planning strategies and to enhance the communication between forest management planners and regional planners.

A recombinant anti‐carcinoembryonic antigen immunoreceptor with combined CD3ζ‐CD28 signalling targets T cells from colorectal cancer patients against their tumour cells

Background and aims: The prognosis of metastatic colorectal cancer is still poor, raising the need for alternative therapeutic approaches, particularly by manipulating the antitumour immune response. Advanced tumour stages, however, are frequently accompanied by functional T cell defects which may be critical for a T cell based anticancer immunotherapy. The aim of this study was to address whether T cells from colorectal cancer patients with advanced tumour stages can be specifically antigen activated against their autologous tumour cells. Methods: T cells were isolated from colorectal cancer patients and retrovirally transduced to express a recombinant immunoreceptor that has an extracellular binding domain for carcinoembryonic antigen (CEA) and an intracellular CD3ζ signalling domain with and without CD28 costimulation for T cell activation. Results: Peripheral blood T cells from colorectal cancer patients were successfully engineered to express the anti-CEA immunoreceptor on the cell surface. On coincubation with autologous CEA+ tumour cells, T cells with anti-CEA immunoreceptor are specifically activated to secrete interferon γ (IFN-γ) and to lyse autologous tumour cells whereas T cells without immunoreceptor are not. T cells equipped with combined CD3ζ-CD28 signalling receptor are more efficiently activated to secrete IFN-γ compared with T cells with CD3ζ signalling receptor. Induction of interleukin 2 secretion on targeting towards autologous tumour cells requires triggering of T cells by the CD3ζ-CD28 costimulatory receptor. Conclusions: T cells from advanced colorectal cancer patients can be tumour specifically activated with high efficiency by engraftment with a combined CD3ζ-CD28 immunoreceptor to break tolerance against autologous tumour cells.

How to better consider sectoral planning information in regional planning: example afforestation and forest conversion

This paper presents, by means of a case study, an approach for how to make use of sectoral planning information on forestry in regional planning. Exemplary issues addressed in this study were, first, how to evaluate the conversion of existing forests and, second, afforestation on agricultural sites, regarding the impact of these strategies on the provision of ecosystem services at a regional scale. We demonstrate that the conversion scenarios planned by the state forest administration have only a minor impact at the regional scale because the proportion of forests is too small. As a consequence, recommendations for regional planning were to: (a) considerably increase the planned afforestation areas under consideration of the locally suitable future forest ecosystem types; and (b) concentrate preference areas for afforestation along corridors, which augment, at most, the additional benefits provided by connecting the biotopes at the landscape level.

Do You Have 5 Minutes To Spare? – The Challenges Of Stakeholder Processes In Ecosystem Services Studies

Operationalization of the ecosystem services (ES) concept for improved natural resource management and decision support cannot, thus far, be rated as satisfactory. Participation of stakeholders is still a major methodical and conceptual challenge for implementing ES. Therefore, we conducted an online survey and a literature analysis to identify benefits and challenges of the application of ES in participatory processes. The results show that the purpose of stakeholder engagement is very diverse as a result of varying objectives, spatial scales and institutional levels of analysis. The complexity, terminology and (lacking) coherent classification of ES are pivotal aspects that should be accounted for in the design of studies to improve stakeholder participation. Although limitations of time and financial resources are bigger challenges than ES related ones, tailoring communication strategies and information for different stakeholder groups are of major importance for the success of ES studies. Results support the view that the potential benefits of applying ES, e.g., consensus finding, and development of integrated solutions, cannot be realized consistently across the different spatial scales and decision-making levels. Focusing on stakeholder processes represents a means to increase the relevance, reliability and impact of study results and to move participation in ES research from theory to reality.

A recombinant anti-CEA immunoreceptor with combined CD3zeta - CD28 signalling targets T cells from colorectal cancer patients against their tumour cells

Background and aims: The prognosis of metastatic colorectal cancer is still poor, raising the need for alternative therapeutic approaches, particularly by manipulating the antitumour immune response. Advanced tumour stages, however, are frequently accompanied by functional T cell defects which may be critical for a T cell based anticancer immunotherapy. The aim of this study was to address whether T cells from colorectal cancer patients with advanced tumour stages can be specifically antigen activated against their autologous tumour cells. Methods: T cells were isolated from colorectal cancer patients and retrovirally transduced to express a recombinant immunoreceptor that has an extracellular binding domain for carcinoembryonic antigen (CEA) and an intracellular CD3ζ signalling domain with and without CD28 costimulation for T cell activation. Results: Peripheral blood T cells from colorectal cancer patients were successfully engineered to express the anti-CEA immunoreceptor on the cell surface. On coincubation with autologous CEA+ tumour cells, T cells with anti-CEA immunoreceptor are specifically activated to secrete interferon γ (IFN-γ) and to lyse autologous tumour cells whereas T cells without immunoreceptor are not. T cells equipped with combined CD3ζ-CD28 signalling receptor are more efficiently activated to secrete IFN-γ compared with T cells with CD3ζ signalling receptor. Induction of interleukin 2 secretion on targeting towards autologous tumour cells requires triggering of T cells by the CD3ζ-CD28 costimulatory receptor. Conclusions: T cells from advanced colorectal cancer patients can be tumour specifically activated with high efficiency by engraftment with a combined CD3ζ-CD28 immunoreceptor to break tolerance against autologous tumour cells.

SAP and SLAM expression in anti-CD3 activated lymphocytes correlates with cytotoxic activity

Signalling lymphocyte activation molecule (SLAM)‐associated protein (SAP) is a small protein that is mutant in humans with X‐linked lymphoproliferative (XLP) disease. Patients with XLP disease are affected by fatal EBV infection and malignant B‐cell lymphomas. The increased risk for B‐cell lymphomas is suggested to result from impaired immunosurveillance of B‐cell proliferation by T cells. In this study, we investigated the role of SLAM and SAP in activation of effector cells with cytotoxic activity, cytokine‐induced killer (CIK) cells, which are generated by non‐specific stimulation of the TCR and addition of exogenous IL‐2. Agonistic TCR activation 1 day after preparation (day +1) resulted in cell activation, with a peak of SLAM on day +6 visible at both the protein and mRNA level as well as membrane detectable SLAM. This increase in SLAM expression correlated significantly with SAP expression at the mRNA level as well as at the protein level. Cytotoxic activity peaked 1 day after the observed SAP and SLAM peaks. At that point in time, IL‐10 secretion, which was high during the early days of culture, decreased. In conclusion, activation of peripheral blood cells with agonistic anti‐CD3 antibody and exogenous IL‐2, as used for generation of CIK cells, results in significant SLAM and SAP activation 5 days after TCR stimulation. This peak correlates with cytotoxic activity against tumour cells. Expression of SLAM and SAP seems to be important in the activation of cytotoxic effector cells.

Anti-tumoral capabilities of effector cells after IFN-α or CpG-motif treatment of cocultured dendritic cells

Abstract.IntroductionEx vivo expansion of monocyte-derived dendritic cells (mDCs) and subsequent coculture with autologous cytokine-induced killer (CIK) cells is an established system to create specific and non-specific anti-tumoral immunity. mDCs constitute the most frequently applied DC subset in clinical studies. One recently published approach to optimize the immunological functions of the DC/CIK cell system is the replacement of interleukin (IL)-4 by interferon (IFN)-α in the maturation process of the DCs.Materials and MethodsThe expressions of relevant surface antigens of IL-4-DCs and IFNα-DCs by flow cytometry and the anti-tumoral activation of effector cells cocultured with both types of DCs using cytotoxicity assays were compared. In addition, short-term coculture experiments with both types of DCs and IFNγ-LAK effector cells were performed and compared with standard CIK cell coculture experiments.ResultsRegarding the expressions of functionally relevant surface markers, no differences could be detected for CD80, CD83, and HLA-DR between IFNα-DCs and IL-4-DCs, whereas the mean fluorescence intensities of CD40, CD86, CD54, and HLA-ABC were decreased and the expression of CD14 was increased for IFNγ-DCs. Moreover, no enhancement of cytotoxicity of cocultured CIK cells against tumor cell lines (A498 and SW480) was detected by the use of IFNα-DCs. Additionally, coculture experiments with IFNγ-LAK cells were performed and unexpectedly higher lysis rates in comparison with the established IL-4-DC/CIK coculture model was observed. Early incubation of the mDCs with several CpG-ODNs failed to increase the anti-tumoral cytotoxicity of the cocultured IFNγ-LAK cells.ConclusionsThese results demonstrate that in the mDC/CIK cell system, IFNα-DCs are not superior in inducing anti-tumoral cytotoxicity and even moderately inferior regarding the expression of functionally relevant surface markers compared with IL-4-DCs.

Pancreas carcinoma antigen fused to invariant chain elicits T-cell response and tumor growth inhibition.

Objectives: The major histocompatibility complex class II chaperone invariant chain (Ii) is widely used as a carrier for inserted antigenic sequences and their introduction into the class II processing pathway. The tumor-associated antigen core 2&bgr; 1,6 N-acetylglucosaminyltransferase (C2GnT), a glycosyltransferase present in human pancreatic tumor cells, is not expressed by normal pancreatic tissues. Methods: A set of expression vectors was engineered where the class II binding region of Ii was replaced by C2GnT-derived sequences. We investigated in vitro whether dendritic cells transfected with Ii-C2GnT constructs were capable to stimulate proliferation of CD4+ T cells. We also tested whether vaccination with Ii-C2GnT would protect mice from tumor development. Results: Invariant chain-C2GnT fusion proteins bind to human DR1, DR3, DR4 and to mouse I-Ab molecules. Our results demonstrate that the plasmid DNA encoding the C2GnT epitope embedded in Ii induces tumor-specific T-cell responses. Mice immunized with the Ii constructs showed reduced growth of Panc02 pancreatic tumor cells. Conclusions: Therefore, Ii clipped with the tumor-associated antigen C2GnT shows promise for the treatment of pancreatic cancer.

Challenges and opportunities of ecosystem service integration into land use planning in West Africa – an implementation framework

ABSTRACT Despite the benefit accrued from integrating ecosystem service (ES) concepts into modern land use planning (LUP) practices, approaches to mainstream the concept in West Africa remains a challenge. The objective of this paper is to develop a framework for integrating ESs into the LUP. We achieved this by using content analysis to search for ES keywords in land use planning policies and act (LUPPA) and to identify existing approaches for mainstreaming the ES approach using Ghana and Nigeria as case-study countries. Following, the SWOT analysis was used to highlight key strength and opportunities of the existing LUPPA, and the benefits the ES concept could offer to increase these strength and opportunities while uncovering the threats to the concepts application in the study location. We suggest the adoption of a transdisciplinary planning approach which integrates strategic environmental assessment and participatory planning and geographic information systems (GIS) approaches, and human resource capacity training of all relevant actors and stakeholders in the planning process on the principles and overall benefits of the ES concept as the way forward. Our framework was developed on the basis of these recommendations for adoption. EDITED BY Davide Geneletti