Dendritic/antigen presenting cell mediated provision of T-cell receptor gamma delta (TCRγδ) expressing cells contributes to improving antileukemic reactions ex vivo

T-cell receptor gamma delta (TCRγδ) expressing T-cells are known to mediate an MHC-independent immune response and could therefore qualify for immune therapies. We examined the influence of dendritic cells(DC)/antigen presenting cell (APC) generated from blast-containing whole blood (WB) samples from AML and MDS patients on the provision of (leukemia-specific) TCRγδ expressing T-cells after mixed lymphocyte culture (MLC). Kit-M (granulocyte-macrophage colony-stimulating factor (GM-CSF) + prostaglandin E1 (PGE1)) or Kit-I (GM-CSF + Picibanil) were used to generate leukemia derived APC/DC (DCleu)from WB, which were subsequently used to stimulate T-cell enriched MLC. Immune cell composition and functionality were analysed using degranulation- (DEG), intracellular cytokine- (INTCYT) and cytotoxicity fluorolysis- (CTX) assays. Flow cytometry was used for cell quantification. We found increased frequencies of APCs/DCs and their subtypes after Kit-treatment of healthy and patients´ WB compared to control, as well as an increased stimulation and activation of several types of immune reactive cells after MLC. Higher frequencies of TCRγδ expressing leukemia-specific degranulation and intracellularly cytokine producing T-cells were found. The effect of Kit-M-treatment on frequencies of TCRγδ expressing cells and their degranulation could be correlated with the Kit-M-mediated blast lysis compared to control. We also found higher frequencies of TCRγδ expressing T-cells in AML patients´ samples with an achieved remission (compared to blast persistence) after induction chemotherapy. This might point to APC/DC-mediated effects resulting in the provision of leukemia-specific TCRγδ expressing T-cells: Moreover a quantification of TCRγδ expressing T-cells might contribute to predict prognosis of AML/MDS patients

The diabetic metabolism and its influence to human CD34+-Progenitorcells

Die Arteriosklerose stellt einen wichtigen Mortalitätsfaktor für Diabetiker dar. Im Vergleich zu Gesunden beginnt die Arteriosklerose bei Diabetikern früher verläuft aggressiver. Ein wichtiger Teil des Pathomechanismus der Arteriosklerose ist die Schaumzellentstehung. In Vorarbeiten zu diesem Projekt konnte gezeigt werden, dass Interaktionen zwischen CD34+-Progenitorzellen und Thrombozyten in der Kokultur mit der Schaumzellbildung endenden. Im Rahmen dieses Projektes wurde untersucht, ob die Herkunft von Progenitorzellen oder von Thrombozyten aus dem diabetogenen Milieu einen proatherogenen Einfluss über eine gesteigerte Schaumzellbildung hat. Grundlage für diese Versuche war die Gewinnung von CD34+-Progenitorzellen gesunder und an Diabetes erkrankter Patienten. Verwendet wurden CD34+-Progenitorzellen aus dem Knochenmark und aus dem Nabelschnurblut, sowie Thrombozyten aus venösem Blut, die in Kokultur bis zur vollständigen Differenzierung der Schaumzellen inkubiert wurden. Zytokinetische Untersuchungen der Kokulturen zeigten eine um 1,6 Tage verlängerte Überlebenszeit der Schaumzellen aus Kulturen diabetischer CD34+-Progenitorzellen und gesunder Thrombozyten. Gleichzeitig konnte zymographisch nachgewiesen werden, dass Kokulturen im diabetogenen Milieu eine signifikant höhere MMP-9 Aktivität aufwiesen während die Oberflächen-Expression des Scavenger-Rezeptors CD68 auf den Schaumzellen der der gesunden Gruppe entsprach. In dieser Arbeit konnte gezeigt werden, dass CD34+-Progenitorzellen im diabetischen Patienten zu einer proatherogen Entwicklung über eine verstärkte und protrahierte Schaumzellbildung beitragen, die mit einer erhöhten inflammatorischen Aktivität des Arteriosklerosegeschehens einhergeht.Atherosclerosis represents an important mortality factor in diabetic patients. Compared to healthy individuals diabetic patients show an earlier onset and a more aggressive development in atherosclerotic lesions. An important part of the atherosclerotic pathomechanism is the genesis of foam cells. Preliminary experiments revealed that the interaction of CD34+-Progenitorcells and platelets results in the generation of foam cells. In this context we examined, whether the origin of progenitor cells or platelets from a diabetic milieu have a proatheroggemtic influence in terms of an enhanced foam cell formation. In this study CD34+-Progenitorcells from healthy and diabetic individuals were used. The CD34+-Progenitorcells used in the cell culture experiments were obtained from cord blood and bone marrow, whereas platelets originated from vein blood. Both cell groups were co-cultured until complete differentiation into foamcells. Cytokinetik analysis of co-cultures consisting of diabetic CD34+-Progenitorcells and healthy platelets showed a prolongation of the foamcell life span of 1.6 days compared to control experiments with healthy CD34+-Progenitorcells and healthy platelets. Additionally zymographic tests showed that co-cultured healthy CD34+-Progenitorcells cells in a diabetic milieu display a significantly enhanced MMP-9 activity while the expression of Scavenger-Rezeptor CD68 showed no difference to the control group. In this study we were able to show, that CD34+-Progenitorcells from diabetic patients show a proatherosclerotic behavior concerning a stronger and faster foamcell formation which leads to a enhanced inflammatory activity in atherosclerosis

CD133, Selectively Targeting the Root of Cancer

Cancer stem cells (CSC) are capable of promoting tumor initiation and self-renewal, two important hallmarks of carcinoma formation. This population comprises a small percentage of the tumor mass and is highly resistant to chemotherapy, causing the most difficult problem in the field of cancer research, drug refractory relapse. Many CSC markers have been reported. One of the most promising and perhaps least ubiquitous is CD133, a membrane-bound pentaspan glycoprotein that is frequently expressed on CSC. There is evidence that directly targeting CD133 with biological drugs might be the most effective way to eliminate CSC. We have investigated two entirely unrelated, but highly effective approaches for selectively targeting CD133. The first involves using a special anti-CD133 single chain variable fragment (scFv) to deliver a catalytic toxin. The second utilizes this same scFv to deliver components of the immune system. In this review, we discuss the development and current status of these CD133 associated biological agents. Together, they show exceptional promise by specific and efficient CSC elimination