Cell sorting for immunological applications.

Project Details

Description

This proposal is joint-application of the Laboratory of Physiology and of Immunology. Both labs need cell-sorting capacity in a number of projects. The Laboratory of Physiology a developed the adeno- and lentiviral transduction of dendritic cells with chlmeric constructs resulting in the presentation of tumor-antigen-derived epitopes in both the context of HLA class I and II. This allows the identification of new class I and class II restricted epitopes recognized by CD8+ and CD4+ T-cells. To clone, expand and test the CD8+ T cells for cytotoxic activity we developed a system of retroviral transduction of EBV-B cells with bicistronic constructs encoding the tumor antigen and a surface expressed marker (ANGFR). This marker allows the enrichment of the transduced cells to homogeneity. The monitoring of the immune response induced by dendritic cell therapy or by protein + adjuvant immunization can be performed by the detection of intracellular cytokines expressed after in vitro restimulation of the in vivo primed T cells. This detection requires also an upgraded flow cytometry facility. In the Laboratoy of Immunology is the need for flow cytometry and sorting essential in a number of studies aiming at a better understanding of the immunobiology of a murine model of multiple myeloma. In a number of studies it has been shown that this model closely mimics the human disease. The role of the expression of IGF1-receptor, clearly demonstrated in a previous study will be further analyzed by separating the IGF1-Rhigh and IGF1-Rlow subpopulations. Also the heterogeneous expression of CD45 (as also observed in human MM) will be studied in detail, as well as the role of metalloproteinases expressed by the tumor cells in their homing characteristics and in the angeogenesis observed in the bone marrow of diseased animals.
AcronymOZR488
StatusFinished
Effective start/end date1/01/0031/12/01

Flemish discipline codes

  • Basic sciences
  • Biological sciences

Keywords

  • multiple myeloma
  • immunotherapy
  • immunobiology