In JoVE (2)
- Um cinética baseada em fluorescência Ca2 + mobilização ensaio para identificar agonistas do Receptor acoplado à proteína G, antagonistas e moduladores alostéricos
- Um ensaio de baseados em citometria de fluxo para identificar compostos que interrompem a ligação do ligante de Chemokine fluorescente-etiquetadas CXC 12 a CXC Chemokine Receptor 4
Other Publications (1)
Articles by Anneleen Van Hout in JoVE
Um cinética baseada em fluorescência Ca2 + mobilização ensaio para identificar agonistas do Receptor acoplado à proteína G, antagonistas e moduladores alostéricos Sandra Claes1, Thomas D'huys1, Anneleen Van Hout1, Dominique Schols1, Tom Van Loy1 1Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven O ensaio de celular descrito destina-se a identificação do receptor do chemokine do CXC 4 (CXCR4)-interação agentes que inibem ou estimulam, competitivos ou qual alostericamente, o intracelular Ca2 + lançamento iniciado pela ativação de CXCR4.
Um ensaio de baseados em citometria de fluxo para identificar compostos que interrompem a ligação do ligante de Chemokine fluorescente-etiquetadas CXC 12 a CXC Chemokine Receptor 4 Geert Schoofs1, Anneleen Van Hout1, Thomas D'huys1, Dominique Schols1, Tom Van Loy1 1Laboratory of Virology and Chemotherapy, Department of Microbiology and Immunology, Rega Institute for Medical Research, KU Leuven Um fluxo de ensaio de ligação celular baseados em citometria é descrito que é usado principalmente como uma ferramenta de triagem para identificar compostos que inibem a ligação de um ligante de chemokine CXC fluorescente etiquetada 12 (CXCL12) para o receptor do chemokine CXC 4 (CXCR4).
Other articles by Anneleen Van Hout on PubMed
Comparison of Cell-based Assays for the Identification and Evaluation of Competitive CXCR4 Inhibitors PloS One. | Pubmed ID: 28410420 The chemokine receptor CXCR4 is activated by its unique chemokine ligand CXCL12 and regulates many physiological and developmental processes such as hematopoietic cell trafficking. CXCR4 is also one of the main co-receptors for human immunodeficiency virus (HIV) entry. Dysfunction of the CXCL12/CXCR4 axis contributes to several human pathologies, including cancer and inflammatory diseases. Consequently, inhibition of CXCR4 activation is recognized as an attractive target for therapeutic intervention. In this regard, numerous agents modifying CXCR4 activity have been evaluated in in vitro experimental studies and pre-clinical models. Here, we evaluated a CXCL12 competition binding assay for its potential as a valuable initial screen for functional and competitive CXCR4 inhibitors. In total, 11 structurally diverse compounds were included in a side-by-side comparison of in vitro CXCR4 cell-based assays, such as CXCL12 competition binding, CXCL12-induced calcium signaling, CXCR4 internalization, CXCL12-guided cell migration and CXCR4-specific HIV-1 replication experiments. Our data indicated that agents that inhibit CXCL12 binding, i.e. the anti-CXCR4 peptide analogs T22, T140 and TC14012 and the small molecule antagonists AMD3100, AMD3465, AMD11070 and IT1t showed inhibitory activity with consistent relative potencies in all further applied CXCR4-related assays. Accordingly, agents exerting no or very weak receptor binding (i.e., CTCE-9908, WZ811, Me6TREN and gambogic acid) showed no or very poor anti-CXCR4 inhibitory activity. Thus, CXCL12 competition binding studies were proven to be highly valuable as an initial screening assay and indicative for the pharmacological and functional profile of competitive CXCR4 antagonists, which will help the design of new potent CXCR4 inhibitors.