Study On The Proliferation Of CFU-GM And Related Regulatory Cytokines In AML Patients

Objective: Both of the CFU-GM assay and MIC technique have prognostic values for untreated acute myelogenous leukemia (AML) patients, but we don't know the relevancy of both prognostic systems and the mechanism on the inhibited growth of myeloid hematopoietic progenitors in vivo in AML patients. In order to clarify these suspicions, we focused on the change of GM-CSF/GM-CSF a receptor and some negative regulators, combined CFU-GM growth characteristics in AML patients, to evaluate their relationship with MIC examination, to detect the expression of GM-CSF /GM-R a /TGF P j/TNF a /LIF and their relations each other, to analyse the effect to CFU-GM growth abnormality and leukemic apoptosis of these cytokines.Methods: In this study, 66 cases of untreated preliminary diagnosised AML patients and 18 cases of control were studied. Their bone marrow CFU-GM were assayed by in vitro hemopoietic progenitor culture, flow cytometry was applied to analyze the expression of GM-CSF a receptor (CD116 antigen) and Fas(CD95)antigen with CD45/SSC gating, ELISA kits were used to detect GM-CSF, solGM-Ra ?TGF ? TNF a and LIF hi patient's plasma.Results: 〤FU-GM growth were inhibited in 66 cases of untreated preliminary diagnosed AML patients, mean colony numbers were 5.8/2X 105MNC, cluster/colony rate reached 12.2 '. 1. Among CFU-GM growth patterns according to Moore's model, I type were 20 cases, CR rate was 25.0%, II type were 12 cases, CR rate was 75.0%, H[ type were 11 cases, CR rate was 27.3%, in IV type, IVA type were 11 cases, CR rate was 90.1%, IV B type were 12 cases, CR rate was 33.3%. The CR rate of IV A+II type CFU-GM growth patterns was higher significantly than that of I +ni+rVB (Pearson Chi-Square x2= 18.796, PO.01) , thus confirmed that CFU-GM growth patterns of AML patients had important value to judge prognosis. Meanwhile, Analysis showed that the prognoses value had good concordance with karyotype abnormal and immunotyping of leukemia.That was, those with high risk karyotypes nearly had nocolony growth and those with middle risk karyotypes had I +m+IVs CFU-GM growth patterns and those with low risk karyotypes had II +!VA CFU-GM growth patterns. In leukemia immunotyping, 75% patients with CD34+ had I +1H+IVB CFU-GM growth patterns, higher significantly than that of II +!VA (P=0.037), also, patients with highest CD13+antigen expression had the IV B CFU-GM patterns and 2/15 patients with CD2 antigen co-expression had the II + IV A CFU-GM patterns. (2) In the research of positive regulators, the expressions of GM-CSF/GM-R a /solGM-R a were all higher than the control. Patients with high levels of solGMR a presents with a distinct clinical picture, these patients had higher white blood cell counts at presentation including myeloid precursors and myeloblasts, also had higher expression of CD34-, CD95^ CD 116 antigen. The patient's plasma GM-CSF were highest in M3( 20.08 ?11.95pg/ml) and lowest (7.93?.10pg/ml) inM6 (PO.01) , and those with higher GM-CSF were CD34"and had low risk karyotypes. Contrast to other FAB subtypes,Ms patients had higher GM-R a (CD 116) expression (40.46 ?7.63%,P<0.01) ,and GM-R a expression was correlation with the expression of CD10> CD 14 antigen; Plasma solGM-R a was highest in M5 patients(9990.92?325.43pg/ml) and lowest in M3(3897.75?651.43 pg/ml) (PO.05) , The expression of GM-R a was correlation with plasma levels of solGM-R a , correlation coefficient is 0.371, PO.01. Among CFU-GM various growth patterns, GM-CSF was highest in IV A type and lowest in III type, solGM-R a was highest hi IV A pattern and lowest in IV B pattern. GM-CSF ^ CD116 and solGM-Ra had no correlation to colony number of CFU-GM. (3)In the research of negative regulators, plasma TGF~Pi> TNF-a and LIF were elevated in AML patients than the control, P value alKO.05. LIF was abnormally elevated in M5 patients; TNF-a was abnormally elevated in M4 and MS patients especially M4; in Me and Ma patients, TGF-pi was higher than others. The plasma levels of LIF in II+IVA type growth pattern is lower than those in I +HI+IVB...