[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].
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.
Wang
Yu-Zhou
YZ
Department of Hematology, Peking Union Medical College Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100730, China. wangyuzhou@hotmail.com
Wu
Yong-Ji
YJ
Zhang
Zhi-Nan
ZN
D000328:Adult; D000368:Aged; D018952:Antigens, CD34 / Q000097:blood; D001854:Bone Marrow Cells / Q000276:immunology* / Q000473:pathology; D018957:CD59 Antigens / Q000097:blood; D002455:Cell Division / Q000187:drug effects; D018920:Coculture Techniques; D017077:Culture Media, Conditioned / Q000494:pharmacology; D003602:Cytotoxicity, Immunologic; D005260:Female; D006457:Hemoglobinuria, Paroxysmal / Q000097:blood* / Q000276:immunology; D006801:Humans; D007371:Interferon-gamma / Q000494:pharmacology; D007376:Interleukin-2 / Q000494:pharmacology; D020014:K562 Cells; D008214:Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000473:pathology; D008297:Male; D008875:Middle Aged
D018952:Antigens, CD34; D018957:CD59 Antigens; D017077:Culture Media, Conditioned; D007376:Interleukin-2; D007371:Interferon-gamma