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  • 2
    Keywords: PEPTIDE ; CELLS ; EXPRESSION ; IN-VITRO ; proliferation ; BLOOD ; CELL ; human ; IN-VIVO ; VITRO ; VIVO ; SITE ; SITES ; DISTINCT ; TISSUE ; LINES ; LIGAND ; RESPONSES ; CUTTING EDGE ; IFN-GAMMA ; INFECTION ; FAMILY ; TISSUES ; CONTRAST ; DENDRITIC CELLS ; T cell ; T cells ; T-CELL ; T-CELLS ; cytokines ; IMMUNE-RESPONSES ; STIMULATION ; EQUIVALENT ; HUMANS ; DIFFERENCE ; ESCHERICHIA-COLI ; MEDIATORS ; LINE ; STIMULI ; COLON-CANCER ; PEPTIDES ; MIGRATION ; PHENOTYPE ; leukocyte ; allogeneic ; immune response ; IMMUNE-RESPONSE ; IMMUNITY ; T lymphocyte ; PERIPHERAL-BLOOD ; PRECURSORS ; AUSTRALIA ; CD34(+) PROGENITOR CELLS ; chemokine ; CTL ; HUMAN DENDRITIC CELLS ; HUMAN PERIPHERAL-BLOOD ; INTERFERON-GAMMA ; LANGERHANS CELLS ; MONOCYTE ; OF-FUNCTION ; SUBSETS ; TOLL-LIKE RECEPTORS
    Abstract: Dendritic cells (DCs) are a family of leukocytes that initiate T- and B-cell immunity against pathogens. Migration of antigen-loaded DCs from sites of infection into draining lymphoid tissues is fundamental to the priming of T-cell immune responses. In humans, the major peripheral blood DC (PBDC) types, CD1c(+) DCs and interleukin 3 receptor-positive (IL-3R(+)) plasmacytoid DCs, are significantly expanded in vivo with the use of Flt3 ligand (FL). DC-like cells can also be generated from monocyte precursors (MoDCs). A detailed comparison of the functional potential of these types of DCs (in an autologous setting) has yet to be reported. Here, we compared the functional capacity of FL-expanded CD1c(+) PBDCs with autologous MoDCs in response to 3 different classes of stimuli: (1) proinflammatory mediators, (2) soluble CD40 ligand trimer (CD40L), and (3) intact bacteria (Escherichia coli). Significant differences in functional capacities were found with respect to changes in phenotype, migratory capacity, cytokine secretion, and T-cell stimulation. MoDCs required specific stimuli for the expression of functions. They responded vigorously to CD40L or E coli, expressing cytokines known to regulate interferon-gamma (IFN-gamma) in T cells (IL-12p70, IL-18, and IL-23), but required prostaglandin E-2 (PGE(2)) during stimulation to migrate to chemokines. In contrast, PBDCs matured in response to minimal stimulation, rapidly acquired migratory function in the absence of PGE(2)-containIng stimuli, and were low cytokine producers. Interestingly, both types of DCs were equivalent with respect to stimulation of allogeneic T-cell proliferation and presentation of peptides to cytotoxic T lymphocyte (CTL) lines. These distinct differences are of particular importance when considering the choice of DC types for clinical applications
    Type of Publication: Journal article published
    PubMed ID: 12738673
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  • 3
    Keywords: brain ; RECEPTOR ; CELLS ; EXPRESSION ; CELL ; Germany ; MODEL ; MODELS ; SYSTEM ; VISUALIZATION ; liver ; GENE ; PROTEIN ; EFFICIENCY ; TISSUE ; MICE ; MECHANISM ; recombination ; TISSUES ; mechanisms ; FREQUENCY ; BONE-MARROW ; ACID ; NUCLEIC-ACIDS ; MOUSE ; IDENTIFICATION ; PROGRESSION ; PROMOTER ; ELEMENTS ; MUTATION ; FUSION ; WILD-TYPE ; MUTATIONS ; IMMUNITY ; MOUSE MODEL ; HEMATOPOIETIC PROGENITOR CELLS ; REGULATORY ELEMENTS ; EMBRYONIC STEM-CELLS ; CRE-MEDIATED RECOMBINATION ; HIGH-AFFINITY RECEPTORS ; HUMAN ERYTHROPOIETIN RECEPTOR ; REPORTER STRAIN
    Abstract: Hematologic disorders can be caused by sporadic or inherited mutations. However, the molecular mechanisms that lead to pathogenicity are only partially understood. An accurate method to generate mouse models is conditional gene manipulation facilitated by the Cre-loxP recombination system. To enable identification and genomic manipulation of erythroid progenitor cells, we established a knock-in mouse model (ErGFPcre) that expresses an improved GFPcre fusion protein controlled by the endogenous erythropoietin receptor (EpoR) promoter. We show that ErGFPcre mice enable the identification of GFP-positive erythroid progenitor cells and the highly specific genomic manipulation of the erythroid lineage. Analysis of GFP-positive erythroid progenitor cells suggests a developmental switch in lineage progression from the hematopoietic stem cell compartment to early erythroid progenitor cells that are stem cell antigen-1-negative (Sca-(1-)) and c-kit(high). Within the hematopoietic system, Cre-mediated recombination is limited to erythroid progenitor cells and occurs in the adult bone marrow at a frequency of up to 80% and in the fetal liver with an efficiency close to 100%. Differential transcriptional activity of the wild-type and the knock-in locus was observed in nonhematopoietic tissues. Thus, our ErGFPcre mouse model could promote the identification of regulatory elements controlling nonhematopoietic EpoR expression and facilitates the characterization and genomic manipulation of erythroid progenitor cells
    Type of Publication: Journal article published
    PubMed ID: 15090451
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  • 4
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    Blood 104 (9), 2988-2989 
    Keywords: CELLS ; EXPRESSION ; TISSUE ; MICE ; MOUSE ; TRANSGENE EXPRESSION ; LINEAGE
    Type of Publication: Journal article published
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  • 5
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    Blood 106 (7), 2228-2229 
    Keywords: CELLS ; EXPRESSION ; BLOOD ; INFECTION ; ANTIGEN ; DENDRITIC CELLS ; T-CELL ; T-CELLS ; IMMUNITY ; CTL ; CAPACITY ; ENZYME ; function ; INDOLEAMINE 2,3-DIOXYGENASE ; TRYPTOPHAN CATABOLISM
    Abstract: Indoleamine 2,3-dioxygenase, an enzyme induced in antigen presenting cells by specific proinflammatory mediators such as prostaglandin E-2 (PGE(2)) or viral infection, can inactivate specific T-cell functions
    Type of Publication: Journal article published
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  • 6
    Keywords: CELLS ; BLOOD ; CELL ; Germany ; human ; POPULATION ; MICE ; PATIENT ; TRANSPLANTATION ; FLOW ; INJECTION ; BONE-MARROW ; MOUSE ; leukemia ; STEM-CELLS ; PHENOTYPE ; REVEALS ; PERIPHERAL-BLOOD ; HEMATOPOIETIC STEM-CELLS ; RE ; CAPACITY ; stem cells ; RECOVERY ; REPOPULATING CELLS ; peripheral blood ; stem cell ; STEM-CELL ; FETAL LIVER ; INTRAFEMORAL TRANSPLANTATION ; LYMPHOID PROGENITOR ; RECONSTITUTION ; SHORT-TERM ; UMBILICAL-CORD BLOOD
    Abstract: Clinical use of purified hematopoietic stem cells in myeloablated patients requires co-transplantation of short-term repopulating cells (STRCs) to ensure timely count recovery. Here, we investigated the flow fluorescence-based side population (SP) phenotype of mobilized human peripheral blood (mPB) cells that rapidly repopulate the highly permissive nonobese diabetic/severe combined immunodeficient (NOD/SCID)-beta 2 microglobulin(-/-) mouse. No SP cells from this source regenerated detectable progeny in these mice before 8 weeks, although by 12 weeks human B-lymphoid cells were seen in some recipients of SP mPB cells. All myeloid reconstituting activity, including that seen within 3 weeks after transplantation, was associated with the non-SP fraction. Isolation of SP cells depletes human mPB of the rapid myeloid reconstitution capacity provided by myeloid-restricted STRCs which are vital for early hematologic recovery in clinical transplant recipients
    Type of Publication: Journal article published
    PubMed ID: 16735598
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  • 7
    Keywords: APOPTOSIS ; CELLS ; EXPRESSION ; SURVIVAL ; BLOOD ; CELL ; Germany ; human ; IN-VIVO ; VIVO ; DEATH ; POPULATION ; GENE ; microarray ; MONOCLONAL-ANTIBODY ; CUTTING EDGE ; MARKER ; CONTRAST ; DENDRITIC CELLS ; T cell ; T cells ; T-CELL ; T-CELLS ; FLOW ; MEMORY ; antibodies ; antibody ; HUMANS ; resistance ; CD95 ligand ; CELL-DEATH ; UP-REGULATION ; RATES ; MARKERS ; MONOCLONAL-ANTIBODIES ; POPULATIONS ; INDIVIDUALS ; CHILDREN ; FLOW-CYTOMETRY ; PERIPHERAL-BLOOD ; MULTIPLE-SCLEROSIS ; HIGH-LEVEL ; CD95 ; cord blood ; INCREASE ; FOXP3 ; LEVEL ; monoclonal antibodies ; RESISTANT ; in vivo ; regulatory T cells ; peripheral blood ; regulatory T cell ; CD4(+)CD25(+) ; CD95L ; CORD ; NEWBORN
    Abstract: Most CD4(+)CD25(hi)FOXP3(+) regulatory T cells (T-regs) from adult peripheral blood express high levels of CD45RO and CD95 and are prone to CD95L-mediated apoptosis in contrast to conventional T cells (T-convs). However, a T-reg subpopulation remained consistently apoptosis resistant. Gene microarray and 6-color flow cytometry analysis including FOXP3 revealed an increase in naive T-cell markers on the CD95L-resistant T-regs compared with most T-regs. In contrast to T-regs found in adult humans, most CD4(+)CD25(+)FOXP3(+) T cells found in cord blood are naive and exhibit low CD95 expression. Furthermore, most of these newborn T-reg are not sensitive toward CD95L similar to naive T-regs from adult individuals. After short stimulation with anti-CD3/CD28 monoclonal antibodies (mAbs), cord blood T-regs strongly up-regulated CD95 and were sensitized toward CD95L. This functional change was paralleled by a rapid up-regulation of memory T-cell markers on cord blood T-regs that are frequently found on adult memory T-reg. In summary, we show a clear functional difference between naive and memory Tregs that could result in different survival rates of those 2 cell populations in vivo. This new observation could be crucial for the planning of therapeutic application of T-reg
    Type of Publication: Journal article published
    PubMed ID: 16868256
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  • 8
    Keywords: SURVIVAL ; Germany ; MODEL ; HYBRIDIZATION ; PATIENT ; IMPACT ; INDUCTION ; treatment ; TRIAL ; IN-SITU ; cytogenetics ; AGE ; chemotherapy ; leukemia ; ABERRATIONS ; PROGNOSTIC-FACTORS ; HIGH-RISK ; PARAMETERS ; PROGNOSTIC-SIGNIFICANCE ; SELECTION ; ABNORMALITIES ; FLUORESCENCE ; ACUTE PROMYELOCYTIC LEUKEMIA ; POSTREMISSION THERAPY ; TRANS-RETINOIC ACID ; in situ hybridization ; PROGNOSTIC-FACTOR ; HIGH-DOSE CYTARABINE ; HISTONE ACETYLATION ; multivariate analysis ; SUBGROUPS ; COOPERATIVE-ONCOLOGY-GROUP ; CORE BINDING ; CUMULATIVE INCIDENCE ; GROUP-B
    Abstract: To assess the prognostic impact of cytogenetics in elderly patients with acute myeloid leukemia (AML) receiving intensive induction and consolidation treatment according to a single protocol specifically designed for patients above age 60, pretreatment samples from 361 patients registered for the AML HD98-B trial of the German-Austrian AML Study Group were analyzed by chromosome banding and fluorescence in situ hybridization, and cytogenetic findings were correlated with outcome. Using a proportional hazards model with backward selection, 3 prognostic subgroups were identified based on the influence of cytogenetic abnormalities on overall survival (OS): low-risk, t(15;17), and inv(16) in 25 of 361 patients (7%); standard-risk, normal karyotype, t(8;21), t(11q23), +8 within a noncomplex karyotype, and +11 within a noncomplex karyotype in 208 of 361 patients (58%); high-risk, all other aberrations in 128 of 361 patients (35%). On multivariate analysis, high-risk cytogenetics (hazard ratio [HR], 2.24) and age above 70 years (HR, 2.34) were independent prognostic factors affecting OS, and stratification according to these parameters demonstrated that a large subgroup of patients (55%), characterized by age 70 or older or high-risk cytogenetics, or both, had very unfavorable treatment results despite intensive chemotherapy. Thus, karyotype and age are major determinants of outcome in elderly patients with AML
    Type of Publication: Journal article published
    PubMed ID: 16840728
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  • 9
    Keywords: RECEPTOR ; APOPTOSIS ; EXPRESSION ; SURVIVAL ; Germany ; DISTINCT ; GENE ; GENE-EXPRESSION ; GENES ; ANTIGEN ; cell cycle ; CYCLE ; PROGNOSTIC-FACTORS ; CD38 EXPRESSION ; SUBSET ; GENOMIC ABERRATIONS ; MUTATION STATUS ; ANTIGEN RECEPTORS ; B-CELL RECEPTOR ; EXPRESSION PATTERNS ; APOPTOSIS-ASSOCIATED GENES ; PATHOMECHANISMS
    Abstract: The mutation status and usage of specific VH genes such as V3-21 and V1-69 are potentially independent pathogenic and prognostic factors in chronic lymphocytic leukemia (CLL). To investigate the role of antigenic stimulation, we analyzed the expression of genes involved in B-cell receptor (BCR) signaling/activation, cell cycle, and apoptosis control in CLL using these specific VH genes compared to VH mutated (VH-MUT) and VH unmutated (VH-UM) CLL not using these VH genes. V3-21 cases showed characteristic expression differences compared to VH-MUT (up: ZAP70 [or ZAP-70]; down: CCND2, P27) and VH-UM (down: PI3K, CCND2, P27, CDK4, BAX) involving several BCR-related genes. Similarly, there was a marked difference between VH unmutated cases using the V1-69 gene and VH-UM (up: FOS; down: BLNK, SYK, CDK4, TP53). Therefore, usage of specific VH genes appears to have a strong influence on the gene expression pattern pointing to antigen recognition and ongoing BCR stimulation as a pathogenic factor in these CLL subgroups
    Type of Publication: Journal article published
    PubMed ID: 16322480
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  • 10
    Keywords: RECEPTOR ; APOPTOSIS ; Germany ; human ; PATHWAY ; PATHWAYS ; LINES ; ACTIVATION ; COMPLEX ; LIGAND ; COMPLEXES ; MECHANISM ; INDUCTION ; CELL-LINES ; CELL-DEATH ; NUMBER ; CELL-LINE ; LINE ; CANCER-CELLS ; CYTOCHROME-C ; cell lines ; DISC ; SIGNALING COMPLEX ; CD95 ; signaling ; PROGRAM ; RE ; FAS ; MEDIATED APOPTOSIS ; SIGNALING COMPLEXES ; ADAPTER MOLECULE
    Abstract: Caspase-2 was reported to be involved in a number of apoptotic pathways triggered by various stimuli. However, the molecular mechanism of procaspase-2 activation in the course of apoptosis remains poorly defined. In this report, we demonstrate that procaspase-2 is recruited to the CD95 (Fas/APO-1) death-inducing signaling complex (DISC) in human T- and B-cell lines. We show that procaspase-2 is activated at the DISC on CD95 stimulation. Despite its presence at the DISC, caspase-2 does not initiate apoptosis on CD95 stimulation in caspase-8-deficient cell lines. Taken together, our data reveal that caspase-2 is activated at the DISC but does not play an initiating role in the CD95-induced apoptosis
    Type of Publication: Journal article published
    PubMed ID: 16822901
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