PRODUCTION OF 2 HUMAN HYBRIDOMAS SECRETING ANTIBODIES TO HLA-DRW11 AND HLA-DRW8+W12 SPECIFICITIES

In this study we describe the production of two human monoclonal antibodies (mAbs) HMP12 and HMP14, that recognize polymorphic HLA-DR specificities. These mAbs have been produced by hybridization of antibody-secreting Epstein-Barr virus-transformed cells with SHM-D33 human-mouse heteromyeloma. By microcytotoxicity assay HMP12 mAb was found to react with all DRw11-positive cells and HMP14 mAb with all cells bearing the DRw8 or the DRw12 specificity. Cytotoxic activity of HMP14 was completely removed after absorption with DRw8- or DRw12-positive cells and unaffected by absorption with cells carrying different DR specificities. The HLA specificity was further analyzed by cytofluorometry on mouse transfectant cells. The reactivity of the two mAbs was correlated with the presence of a particular polymorphic amino acid residue in the DR beta chain and by this approach the epitopes possibly involved in the antibody binding sites were predicted.