Document Type : Original Article
IMMUNOLOGY DEPARTMENT FACULITY OF MEDICINE, TARBIAT MODARRES UNIVERSITY, TEHRAN, IRAN
Introduction: Enrichment of intact peripheral blood dendritic cells (DCs) has been done using rosette and immunomagnetic depletion techniques.
Material and Methods: Peripheral blood mononuclear cells were separated using ficoll density gradient centrifugation and T lymphocytes were depleted through reaction with AET- treated sheep red blood cells. The remaining cells were depleted from lineage marker positive cells using monoclonal antibodies against CD3, CD11b, CD14, CD16, CD19, CD56 and anti- mouse IgG magnetic beads. Flowcytometric analysis was used to determine the purity of the obtained DCs through HLA-DR expression and lack of lineage markers.
Results: The obtained results showed that a significant population of isolated cells (58.5% ± 4.5, n=5) fail to react with anti-lineage marker monoclonal antibodies but express the HLA-DR antigen.
Conclusion: Immunomagnetic depletion is a new method for DCs separation that unlike classic methods, can isolate DCs in intact form. According to our information, this is the first report of DCs separation in Iran and purity of the obtained DCs is comparable with similar studies in other countries. Significant enrichment of intact DCs could facilitate the study of DCs natural functions in different fields such as autoimmunity, cancers and infectious diseases.