Proliferation and anti-tumor activity of lymphocytes and their effect on normal phenotype of autologous bone marrow hematopoietic cells in patients with paroxysmal nocturnal hemoglobinuria / 中国实验血液学杂志

ABSTRACT

There is a hypothesis that paroxysmal nocturnal hemoglobinuria (PNH) hematopoitic stem cells are resistant to the cytotoxic effect of T cells because they lack glycosylphosphatidylinositol (GPI)-linked proteins. The aim of this study is to investigate proliferation and anti-tumor activity of lymphocytes in patients with PNH, and also to assay the effect on normal cell (CD59(+)) phenotype in bone marrow of PNH patient by autologous lymphocytes in vitro. MTT assay for detection of lymphocytes proliferation and its anti-tumor effect was driven to delineate T cell reactive function. The CD34(-) and CD34(+) bone marrow cells (selected by means of immunomagnetic method) as well as unsorted marrow cells in PNH patients were cultured together with autologous CD59(+) or CD59(-) lymphocytes, their cultured supernatant, extrinsic IFN-gamma and IL-2 in a liquid culture system. CD59(+) cells were counted by flow cytometry after 10 day culture in vitro. The results showed that there were no differences in the proliferation ability of lymphocytes between each group controls 1.42 +/- 0.46, unsorted PNH lymphocytes 1.40 +/- 0.35, CD59(-) 1.30 +/- 0.40, and CD59(+) PNH lymphocytes 1.40 +/- 0.42. Anti-tumor effect of lymphocytes declined in PNH patients when compared with control [(50.00 +/- 28.67)% vs. (76.13 +/- 10.15)%]. The proportion of CD59(+) cells diminished significantly after culture with autologous lymphocytes, their supernatant, extrinsic IFN-gamma or IL-2 (P < 0.01) in groups of unsorted, CD34(+) and CD34(-) bone marrow cells. No significant difference was found between groups of CD59(-) and CD59(+) lymphocytes, or CD34(-) and CD34(+) marrow cells. It is concluded that turbulences of immune regulations may be involved in pathogenesis of PNH.