Role of complement regulatory proteins and adherent factors on the differentiation from the stem cells in paroxysmal nocturnal hemoglobinuria

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Expressions of complement regulatory protein, HRF20 have been investigated on the cell membrances of hematopoietic stem cells and their progeny cells in bone marrows from patients with paroxysmal nocturnal hemoglobinuria(PNH). PNH nonphagocytic mononuclear marrow cells were labeled with anti-HRF20 monoclonal antibody and subsequently with FITC-conjugated secondary antibody and then sorted into HRF20-negative and -positive fractions with a FACS, followed by the culturing with them in methylcellulose. CFU-GM colonies and BFU-E bursts were formed in the cultures 10 to 14 days later. These colonies/bursts were plucked up and their progeny cells were analyzed with respect to expression of HRF20. The expression was negative on all of the progeny cells from HRF20-negative progenitors, while the cells from HRF20-positive progenitors revealed both negative and positive expressions of HRF20 on their cell membranes.