Search

Cell therapy with antigen-specific T cells holds promise for various diseases including cancer and viral infections. The powerful enrichment procedure based on major histocompatibility complex (MHC)-tetramers, however, is of limited applicability so far. Therefore, the recently developed cell surface affinity matrix technology that allows direct identification and enrichment of life antigen-specific T cells based on cytokine secretion was evaluated in this respect. To this end, CD8+ T cells directed against the HLA-A*0201-restricted melanoma-associated peptide Melan-A (aa26–35) were generated by combining stimulation of peptide-pulsed autologous dendritic cells (DC) with antigen-independent expansion with anti-CD3/anti-CD28 monoclonal antibodies (MoAb). Antigen-specific cytotoxic T lympocyte (CTL) were detected based on stimulation-induced interferon (IFN)-γ and interleukin (IL)-4 secretion and enriched > 100-fold using the cell surface affinity matrix technology. The resulting IFN-γ- and IL-4-secreting CTL lines contained > 80% and > 70% cytokine positive T cells, respectively. They exhibited a cytotoxic activity against Melan-A expressing target cells that was significantly higher as compared to nonpurified CTL. Direct staining of enriched CTL with HLA-A2-Melan-A-tetramers revealed a high correlation between the results obtained from the cell surface affinity matrix technology and those obtained from tetrameric complexes. Altogether, our study demonstrates that cytokine-driven enrichment based on the cell surface affinity matrix technology enables selective isolation of functionally active antigen-specific CTL that may be used for an adoptive T cell transfer in immunotherapy. [ABSTRACT FROM AUTHOR]

To detect risky eating behavior questionnaires should be economic but at the same time they should fulfill the psychometric quality criteria. Available instruments are too long for the target group (e. g. EDE-Q, 28 items), restricted on primary symptoms (short version of EDI, 23 items) and with minor reliability (e. g. SCOFF and WC-Scale, 5 items each). Using the German version of the Eating Attitudes Test (EAT-26D, which comprises 26 items) in a community sample of 1 331 11-13 year old girls and 906 boys from Thuringia, Germany, we measured a internal consistency of Cronbachs' Alpha=0.85 for girls and 0.78 for boys. In a principal factor analysis, we could replicate the 6-factorial structure of previous studies. A confirmatory factor analysis verified the suitability of the EAT-26D for both, girls and boys. Reducing the EAT-26D on the 3 core-factors leads to an economic 13 item short version with an internal consistency of 0.87 for girls and 0.80 for boys., (© Georg Thieme Verlag KG Stuttgart · New York.)

Objectives: The objectives of this study were to assess cytokine secretion in human renal cell carcinoma (RCC) and to identify cytokines contributing to the immunomodulatory effect of tumor cells., Methods: Cytokine secretion in the supernatant of primary tumor cell cultures (PTCC) and corresponding cell lines (CL) was assayed using ELISA. Tumor cells were characterized by morphology, immunocytochemistry, and flow-cytometric analysis. Tumor-cell-induced T cell activation was determined by coculture of gamma delta and alpha beta T cell clones with tumor CL., Results: We assessed the cytokine secretion of tumor cells from 27 PTCC and their corresponding CL (3/27) of RCC. We found that RCC predominantly produced both pro-inflammatory and T-cell-inhibitory cytokines, such as IL-8, IL-6, GM-CSF, TNF-alpha, IL-10 and TGF-beta 1. CL were adapted to serum-free medium which may prove as a useful tool in future studies of cytokine secretion in RCC. In addition, we used gamma delta and alpha beta T cell clones to assess the immunomodulatory effect of tumor cells from RCC and found that predominantly gamma delta T cells were activated by RCC., Conclusions: Our data suggest that RCC produce large amounts of both pro-inflammatory and T-cell-inhibitory cytokines that potentially could influence the immune response of the host, especially tumor-specific cytotoxic T cells.