TY - JOUR A1 - Bergholz, André A1 - Heymann, Stephan A1 - Schenk, Jörg A. A1 - Freytag, Johann Christoph T1 - Biological sequences integrated: a relational database approach N2 - Over the last decade the modeling and the storage of biological data has been a topic of wide interest for scientists dealing with biological and biomedical research. Currently most data is still stored in text files which leads to data redundancies and file chaos. In this paper we show how to use relational modeling techniques and relational database technology for modeling and storing biological sequence data, i.e. for data maintained in collections like EMBL or SWISS-PROT to better serve the needs for these application domains. For this reason we propose a two step approach. First, we model the structure (and therefore the meaning of the) data using an Entity-Relationship approach. The ER model leads to a clean design of a relational database schema for storing and retrieving the DNA and protein data extracted from various sources. Our approach provides the clean basis for building complex biological applications that are more amenable to changes and software ports than their file-base counterparts. Y1 - 2001 UR - http://www.springerlink.com/app/home/ contribution.asp?wasp=161c4c19086a4dceac9312b46e5f2348&referrer=parent&backto=issue,1,5;journal,14,29;linkingpublicationr esults,1:102835,1 ER - TY - JOUR A1 - Pecher, Gabriele A1 - Harnack, U. A1 - Gunther, M. A1 - Hummel, M. A1 - Fichtner, I. A1 - Schenk, Jörg A. T1 - Generation of an immortalized human CD4+ T cell clone inhibiting tumor growth in mice. N2 - Tumor antigen-specific T cell clones represent a useful tool in tumor immunology; however, their long-term culture is limited. To generate an immortalized cytotoxic T cell clone against the human tumor antigen mucin, we exposed a previously generated T cell culture to Herpesvirus saimiri. We obtained an immortalized human CD4+ T cell clone, termed SITAM. Clonality of these cells was shown by analysis of the alpha/beta-T cell receptor (TCR) repertoire. Cytolytic activity was demonstrated against several mucin-expressing tumor cell lines and could not be detected against non-mucin-expressing cells. SITAM cells maintained their features stably for 2 years. Furthermore, growth of the tumor cell line Capan-2 in NOD/SCID mice was inhibited when SITAM cells were coinjected subcutaneously with tumor cells. SITAM cells provide an unlimited source of clonal T cells for analysis of tumor recognition and may be of help in TCR-targeted immunotherapy. Y1 - 2001 UR - http://www.sciencedirect.com/science?_ob=ArticleURL&_udi=B6WBK-45S4JD7- VM&_coverDate=05%2F18%2F2001&_alid=268965202&_rdoc=1&_fmt=&_orig=search&_qd=1&_cdi=6713&_sort=d&view=c&_acct=C000053886&_ v ER - TY - JOUR A1 - Pecher, Gabriele A1 - Schirrmann, Thomas A1 - Kaiser, Lothar A1 - Schenk, Jörg A. T1 - Efficient cryopreservation of dendritic cells transfected with cDNA of a tumour antigen for clinical application N2 - Dendritic cells (DCs) are the most potent antigen-presenting cells of the immune system and are currently being investigated in clinical applications as cancer vaccines. An efficient cryopreservation method would greatly contribute to their use in clinical trials. We have established a method for freezing of DCs derived from peripheral blood mononuclear cells using the plasma expander Gelifundol. This enabled us to reduce the concentration of the toxic DMSO to 5%. The method could be performed without the addition of fetal calf serum or any other serum. After freezing, the viability of the DCs was 90%. The cells exhibited all the phenotypic characteristics (CD11c+, HLA-DR+, CD80+, CD83+, CD86+) of DCs, as tested by flow cytometry. Cells transfected with cDNA for the tumour antigen mucin expressed this protein on their surfaces in the same manner as before freezing. The stimulating capacity of a mixed lymphocyte culture was also preserved. These findings offer an efficient method for the cryopreservation of DCs for use in clinical trials. Y1 - 2001 UR - http://www.babonline.org/bab/034/0161/bab0340161.htm ER - TY - JOUR A1 - Pecher, Gabriele A1 - Spahn, Gunter A1 - Schirrmann, Thomas A1 - Kulbe, Hagen A1 - Ziegner, Maja A1 - Schenk, Jörg A. A1 - Sandig, Volker T1 - Mucin gene (MUC1) transfer into human dendritic cells by cationic liposomes and recombinant adenovirus N2 - BACKGROUND: Dendritic cells (DC) as antigen presenting cells play an important role in immunotherapy of cancer. Mucin, encoded by the gene MUC1, is a human tumor antigen expressed in breast, pancreatic and ovarian cancers. Therefore, MUC1-transfected DC would be an attractive tool in constructing cancer vaccines. MATERIALS AND METHODS: Using two different cationic liposome preparations and, for comparison, a recombinant adenovirus expressing mucin, we tested the efficiency of mucin gene transfer into DC by flow cytometry. We investigated if these transfected DC were able to specifically stimulate autologous peripheral blood lymphocytes (PBL) from healthy donors. RESULTS: Flow cytometry revealed that 5-20% of DC transfected with liposomes Lipofectin and 20-40% of DC transduced with adenovirus expressed the relevant mucin epitopes. The expression of mucin on DC was similar to the expression of mucin found on carcinoma cells. After antigen uptake, DC specifically stimulated autologous PBL. CONCLUSION: We have shown that cationic liposomal gene transfer into human DC was feasible. We could obtain antigen specific stimulation of PBL at a similar rate as with adenoviral MUC1-transduced DC. Y1 - 2001 SN - 0250-7005 ER -