Showing papers by "Steffen Goletz published in 2004"
TL;DR: A tumor-specific MUC1 epitope is defined comprising the ...PDTRP... sequence in a particular conformation essentially determined by O-glycosylation at its threonine with either GalNAcalpha1 or a related short glycan.
Abstract: Glycosylation determines essential biological functions of epithelial mucins in health and disease. We report on the influence of glycosylation of the immunodominant DTR motif of MUC1 on its antigenicity. Sets of novel glycopeptides were synthesized that enabled us to examine sole and combined effects of peptide length (number of repeats) and O-glycosylation with GalNAc at the DTR motif on the binding patterns of 22 monoclonal antibodies recognizing this motif. In case of unglycosylated peptides almost all antibodies bound better to multiple MUC1 tandem repeats. Glycosylation at the DTR led to enhanced binding in 11 cases, whereas 10 antibodies were not influenced in binding, and one was inhibited. In nine of the former cases both length and DTR glycosylation were additive in their influence on antibody binding, suggesting that both effects are different. Improved binding to the glycosylated DTR motif was exclusively found with antibodies generated against tumor-derived MUC1. Based on these data a tumor-specific MUC1 epitope is defined comprising the ...PDTRP... sequence in a particular conformation essentially determined by O-glycosylation at its threonine with either GalNAcalpha1 or a related short glycan. The results can find application in the field of MUC1-based immunotherapy.
98 citations
TL;DR: It is postulate that scFv phagemid library formats with short linkers may, in general, be advantageous in selections for the generation of scFvs against carbohydrate epitopes or other epitopes associated with low intrinsic affinity per binding site), and expect that they will be superior in applications for diagnosis or therapy.
44 citations
TL;DR: The present communication provides a fast, cheap, and efficient separation method by precipitating the DNA from a phosphate buffered solution with manganese chloride, especially valuable for antibodies which lose bioactivity by interactions with chromatographic matrices.
5 citations
Journal Article•
TL;DR: The data indicate that NMDC represent a ready and standardized source of semi-allogeneic DC for immunotherapeutic approaches aiming at augmented CTL functions and can be primed by NMDC with equivalent efficiency to autologous MoDC.
Abstract: 1285 We have recently shown that dendritic cells can be generated from the HLA-A2+ CD34+ human acute myeloid leukemia cell line MUTZ-3. Upon stimulation with GM-CSF, IL-4 and TNF-α, MUTZ-3 precursor cells acquire a dendritic cell phenotype consistent with conventional interstitial DCs, displaying the full range of functional antigen processing and presentation pathways. These DC are hereafter referred to as NMDC. In our current experiments, the capacity of NMDC to induce tumor-specific CTL in vitro was examined and compared to the induction capacity of autologous monocyte-derived dendritic cells (MoDC). To this end, HLA-A*0201+/CD8β+ CTL precursors from healthy donors were weekly stimulated with either autologous MoDC or NMDC, both pulsed with the tumor-associated A2-binding peptides hTERT988Y or CEA571-579 (Cap-1). Equivalent CTL priming efficiencies were observed for MoDC and NMDC. After isolation by CD8/tetramer-guided flow sorting and cloning by limiting dilution, the functional capacity of the obtained CTL clones generated was determined. CTL clones could be identified with CEA571-579 specific cytolytic activity. With an intracellular IFN-γ staining procedure we were able to show that these clones recognized the naturally processed epitope on KATO-3, an HLA-A*0201+/CEA+ tumor cell line. We conclude that functional tumor-specific CTL can be primed by NMDC with equivalent efficiency to autologous MoDC. Our data indicate that NMDC represent a ready and standardized source of semi-allogeneic DC for immunotherapeutic approaches aiming at augmented CTL functions.
1 citations
Patent•
23 Jan 2004
TL;DR: The authors concerne des molecules de reconnaissance dirigees contre les tumeurs, les compositions pharmaceutiques contenant ces molecules de re reconnaissance, des procedes de production of ces de reconnaissance and l'utilisation de ces dernieres for le diagnostic and la therapie de maladies tumorales.
Abstract: L'invention concerne des molecules de reconnaissance dirigees contre les tumeurs, les compositions pharmaceutiques contenant ces molecules de reconnaissance, des procedes de production de ces molecules de reconnaissance et l'utilisation de ces dernieres pour le diagnostic et la therapie de maladies tumorales.
Patent•
18 Aug 2004
TL;DR: The authors concerne egalement des compositions, de preference des compositions pharmaceutiques ou vaccinales qui comprennent les lignees cellulaires, le lysat, les molecules, le melange de molecules ou les cellules dendritiques of the present patent.
Abstract: La presente invention concerne une lignee cellulaire selectionnee dans le groupe comprenant : (a) une lignee cellulaire appelee NM-F9 presentant le numero d'ordre DSMZ, DSM ACC2606 ; (b) une lignee cellulaire appelee NM-D4 presentant le numero d'ordre DSMZ, DSM ACC2605 ; et des sous-clones de (a) ou (b). En outre, la presente invention concerne un lysat des lignees cellulaires ou une molecule ou un melange de molecules obtenu a partir desdites lignees cellulaires ainsi que des cellules dendritiques chargees dudit lysat, co-cultivees ou fusionnees avec des cellules desdites lignees cellulaires, ou une molecule ou un melange de molecules obtenu a partir desdites lignees cellulaires de la presente invention. L'invention concerne egalement des compositions, de preference des compositions pharmaceutiques ou vaccinales qui comprennent les lignees cellulaires, le lysat, les molecules, le melange de molecules ou les cellules dendritiques de la presente invention. Dans un autre mode de realisation, la presente invention concerne des methodes de production des compositions susmentionnees. En outre, l'invention concerne des methodes et des utilisations de la vaccination dirigee contre ou pour le traitement ou la prevention de cancers et/ou de maladies tumorales.