Poster Group 1 - Genomic Analysis of MHC and Related Genes

In vitro characterisation of alloreactivity induced by isolated HLA antigens of interest is an important tool for the dissection of permissive and non-permissive HLA mismatches, one of the challenges of modern transplantation biology. The use of autologous antigen presenting cells (APCs) transfected with the HLA allele of interest is hampered by the scarce transfectability of professional APCs. In this study, we have circumvented this problem by use of autologous, PHA-activated T cell blasts as APCs. These cells are readily transfected with HLA alleles of interest by retroviral vector mediated gene transfer and subsequent purification of transduced cells by immunobead selection for the cell surface marker NGFr. These T cell blasts were capable of efficient and specific stimulation of alloresponses in mixed lymphocyte cultures The system is currently being used to study in vitro alloreactivity induced by permissive or non-permissive HLA DP 1 a lleles. Another application will be induction of single alloantigen specific regulatory T cells that could be useful for tolerance induction in mismatched haematological stem cell transplantation. A NOVEL METHOD TO DETECT SOLUBLE HLA-G SECRETING CELLS