Characterization of intracellular HLA-DR, DM and DO profile in K562 and HL-60 leukemic cells.

Surface class-II antigen expression fires-up the antigen presentation process and development of immune response. The absence of surface HLA-DR is used in various systems to avoid immune recognition. Most leukemic cells use such mechanism to escape immune surveillance. Here, K562 and HL-60 leukemic cells were examined as to intracellular HLA-DR, -DM and -DO expression, if any. Immunofluorescence scored by UV-microscopy, flow cytometry or confocal microscope analysis detected intracellular pools of HLA-DR, -DO and to a lesser degree HLA-DM, whereas sub-cellular fractionation localized these molecules within endosomes. RT-PCR experiments revealed the presence of HLA-DRalphabeta, HLA-DMalphabeta and HLA-DObeta but not HLA-DOalpha transcripts. Despite the absence of the HLA-DOalpha chain, stable transfectants of K562 with a full length HLA-DObeta-EGFP construct showed that DObeta chain could be translocated to endosomes and form stable complexes with HLA-DR. Such complexes could be responsible for arresting HLA-DR molecules within endosomes, maintaining their surface class-II negative state.