Objective The objectives of the proposed study include: 1- To isolate a set of yeast artificial chromosome (YAC) clones encompassing a region of human chromosome 12 containing several families of known and potentially novel NK cell receptor genes 2- To isolate representative members of each family, map their chromosomal location and analyze their expression in subsets of NK cells at the gene and protein level 3- To study the role of selected members of the individual families in NK cytotoxicity assays and to analyze the intracellular signalling pathways triggered by the receptors 4- To determine the structures on target cells to which these receptors are binding, especially to which extent specific carbohydrates, glycoproteins and/or MHCI haplotypes are the ligands Brief description: A significant fraction of blood leucocytes are large granular lymphocytes. Because of their ability to spontaneously kill certain tumour cells and virally infected cells without prior sensitization, these cells are known as natural killer (NK) cells. They appear to be important for the resistance to growth and metastatic spread of tumours and for natural immunity against viral infections. In addition they have been implicated in bone marrow graft rejection Whereas the antigen-recognising receptors and the corresponding genes of B and T Lymphocytes have been well characterised within the last decade, the precise nature and function of the NK cell receptors mediating the recognition of aberrant cells remains to be elucidated. Several laboratories, including our have recently described and analysed candidate proteins in humans and the rodent system. Most of these potential NK cell receptors can be grouped in several sma families and share common structural features, including a type ll transmembran orientation and a lectin domain in the external part. It is our hypothesis that the expression of a specific array of these lectin type molecules is one of the determinants of functional subsets of NK cells in humans and is also responsible for the observed NK activity in certain T Lymphocyte subsets. This project proposes to delineate first the repertoire of human NK cell receptors from genomic mapping analysis and sequencing efforts. It has already been demonstrated that a region containing several human lectin-type receptor genes locates to chromosome 12 of humans. The available data suggest that similarly to the situation in mice, where homologous families have been shown to map to the isotenic murine chromosome 6, this genomic region contains in addition to the human NKG2 also the recently isolated CD94 and several additional lectin-type gene families. We propose to isolate representative members of the different families, map their location on the chromosome and analyze their expression and function together with the recently described forms of NKG2 and CD94. The expression of selected families or isoforms in NK cell subsets, the role of individual proteins for NK cell effector functions and their ligands on target cells and normal cells will be studied. This should be achieved by the collaboration of six complementary laboratories: One group being involved in the analysis of the NKG2 family (U.Wien) one has recently identified the CD94 gene (U.Madrid) another group will collaborate to isolate the necessary basis of genomic YAC clones and will contribute sophisticated mapping and screening procedures necessary for the intended studies (MPI/Genetik), one laboratory is specialised in carbohydrate biology with a track record in discovering novel saccharide ligands (MRC/Glyco), one will collaborate to test for MHCI recognition by the lectintype molecules contributing its extensive experience in MHCI/NK cell interactions (Karolinska) and one will study intracellular signalling pathways triggered by selected receptors (U.Sapienza). We believe that this partnership is a unique combination of different expertises that will contribute to European competitiveness in the newly developing area of NK cell function and the results obtained could provide the basis for the development of novel approaches to (cellular) immunotherapy of cancer and viral diseases and for the prevention of certain forms of transplant rejection. Fields of science natural sciencesbiological sciencescell biologycell signalingnatural sciencesbiological sciencesbiochemistrybiomoleculesproteinsmedical and health sciencesclinical medicineoncologymedical and health sciencesbasic medicineimmunologyimmunotherapynatural sciencesbiological sciencesgeneticschromosomes Programme(s) FP4-BIOTECH 2 - Specific research, technological development and demonstration programme in the field of biotechnology, 1994-1998 Topic(s) 0501 - Immunology and immunotechnology Call for proposal Data not available Funding Scheme CSC - Cost-sharing contracts Coordinator Universität Wien EU contribution No data Address 59,Brunner Strasse 1235 Wien Austria See on map Total cost No data Participants (5) Sort alphabetically Sort by EU Contribution Expand all Collapse all Imperial College of Science Technology and Medicine United Kingdom EU contribution No data Address Norfolk Place W2 1PG London See on map Total cost No data KAROLINSKA INSTITUTE Sweden EU contribution No data Address 13,Nobels väg 16 171 77 Stockholm See on map Total cost No data MAX-PLANCK-GESELLSCHAFT ZUR FOERDERUNG DER WISSENSCHAFTEN E.V. Germany EU contribution No data Address Ihnestrasse 73 14195 BERLIN See on map Total cost No data UNIVERSIDAD AUTONOMA DE MADRID Spain EU contribution No data Address 62,Diego de Leon 62 28006 MADRID See on map Total cost No data Università degli Studi di Roma "La Sapienza" Italy EU contribution No data Address 324,Viale Regina Elena 00161 Roma See on map Total cost No data
