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Aims: HER2 testing of invasive breast cancer by in-situ hybridization guides therapy decisions. Probing HER2 and centromere of chromosome 17 (cen17) simultaneously is supposed to reveal both a potential HER2 gene amplification and polysomy 17. However, a considerable number of breast cancer patients with quasi polysomy 17 are considered âequivocalâ, which is diagnostically meaningless. Moreover, patients with equivocal/false polysomic tumours are prevented from a potentially beneficial anti-HER2 treatment. Here we evaluated the RAI1, D17S122 and TP53 hybridization markers to indicate true polysomy reliably and to reclassify equivocal samples accurately as HER2-positive. Methods and results: Samples with (n = 82) and without (n = 52) increased cen17 copy numbers and 78 evidently HER2-amplified specimens were analysed using dual and triple marker hybridization probes. Selected putative polysomic samples were subjected to array-based comparative genomic hybridization (aCGH). We found that 37.8% samples with putative polysomy 17 did not show any gain in RAI1, D17S122 or TP53. Accordingly, aCGH revealed evidence for the presence of HER2/cen17 co-amplification rather than for true polysomy in those cases. Reflex testing using alternate 17p markers reclassified up to 56.3% equivocal cases as HER2-positive and the combined assessment of a 17p and 17q locus allows the differentiation of true versus false polysomy. Conclusions: An increased cen17 copy number does not necessarily reflect polysomy, and reflex testing facilitates the reclassification of 'equivocals'. Nevertheless, the prognostic and predictive value of a HER2/cen17 co-amplification versus HER2 gene amplification alone must still be evaluated prospectively.

Hintergrund und Fragestellung: Die vorliegende Studie untersucht, ob es möglich ist durch konventionelle histopathologische Parameter eine ausreichende Darstellung der Tumorbiologie des Mammakarzinoms zu erreichen und sich diese Subtypisierung auf das Überleben auswirkt. Methode: Studiengrundlage[zum vollständigen Text gelangen Sie über die oben angegebene URL], 15. Deutscher Kongress für Versorgungsforschung

Breast cancer is a highly inter- and intra-individual heterogeneous disease that comprises a number of subtypes for which no therapeutic antibodies are available yet. Hence we utilized the Humanized Tumor Mouse (HTM) model and evaluated its capability to generate tumor cell specific human antibodies directed against BC cells. HTM were generated by the co-transplantation of human hematopoietic stem cells and human Her2-positive breast cancer cells into neonatal NSG mice. These mice develop a human immune system in combination with human breast cancer growth. Due to the concurrent transplantation the transfer of MHC-mismatched tumor cells causes an activation of a variety of immune (CD4+, NK, myeloid) cells without evidence for rejection. Histological staining of the spleen of HTM revealed co-localization of human antigen-presenting cells together with human T- and B-cells enabling an MHC-dependent interaction and thereby the generation of T-cell dependent antibody production by B-cells. Overall, HTM were able to generate human tumor-specific antibodies which dose dependently inhibited tumor outgrowth. Western blot analyses revealed that the tumor-specific antibodies generated in HTM recognized antigens which were different from Her2. In conclusion, HTM offer a novel approach to generate complete human monoclonal antibodies that do not require further genetic manipulation (e. g. "humanization"). This might be of particular interest for cancer subtypes with no currently available antibody therapy.

Fragestellung: Aus Lipoaspirat oder Fettgewebe kann mittels enzymatischer Verdauung durch eine Kollagenase und anschließende Zentrifugation die stammzellreiche Stromal Vascular Fraction (SVF) gewonnen werden. Bisher hat sich jedoch weder ein einheitliches Extraktionsverfahren, noch eine gängige[zum vollständigen Text gelangen Sie über die oben angegebene URL], Deutscher Kongress für Orthopädie und Unfallchirurgie (DKOU 2015)

Monoclonal antibodies against the human homologue of mouse macrosialin, CD68, are generally commercialized as markers for human monocytes and macrophages. Indeed, CD68 is considered as a selective marker for human myeloid cells, although several previous immunohistochemical studies indicate that some antibody clones also react with other hematopoietic and non-hematopoietic cell types. The aim of our study was to verify these observations and to evaluate the reliability of CD68 as a macrophage marker.We investigated protein and RNA expression of CD68 in various fibroblast types and carcinoma cell lines as compared to monocytes and macrophages using immunohistochemistry, flow cytometry, and specific RT-PCR. Different monoclonal antibody clones against CD68 were applied including KP-1 and EBM11.As expected, the intensity of immunohistochemical and flow cytometric CD68 staining was dependent on both the antibody clone and the fixation procedure. However, fibroblasts isolated from normal skin, normal breast, breast tumor tissue, and osteoarthritis synovia clearly expressed CD68 protein at levels comparable to macrophages. The specificity of CD68 expression in fibroblasts was verified by RT-PCR which also showed some tumor cell types to express CD68 mRNA.Our findings clearly demonstrate that the expression of CD68 is not restricted to the macrophage lineage. This is highly relevant for experimental and diagnostic purposes, since anti CD68 antibodies cannot be accepted without reservations for the discrimination of myeloid cells and fibroblasts even in paraffin sections after formalin fixation.