TY  - THES
A1  - Schreiber, J.
A1  - Stahn, R.
A1  - Schenk, Jörg A.
A1  - Karsten, U.
A1  - Pecher, Gabriele
T1  - Binding of tumor antigen mucin (MUC1) derived peptides to the heat shock protein DnaK
Y1  - 2000
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  - 
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  -