Subtypes of B lymphocytes in patients with autoimmune hemocytopenia.

OBJECTIVE: To investigate the quantities of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia and the relationship between quantities of CD5+ B lymphocytes and clinical or laboratorial parameters. METHODS: Quantities of CD5+ B lymphocytes in the bone marrow of 14 patients with autoimmune hemolytic anemia (AIHA) or Evans syndrome, 22 immunorelated pancytopenia (IRP) patients, and 10 normal controls were assayed by flow cytometry. The correlation between their clinical or laboratorial parameters and CD5+ B lymphocytes was analyzed. RESULTS: The quantity of CD5+ B lymphocytes of AIHA/Evans syndrome (34.64% +/- 19.81%) or IRP patients (35.81% +/- 16.83%) was significantly higher than that of normal controls (12.00% +/- 1.97%, P < 0.05). However, there was no significant difference between AIHA/Evans syndrome and IRP patients (P > 0.05). In all hemocytopenic patients, the quantity of bone marrow CD5+ B lymphocytes showed significantly negative correlation with serum complement C3 level (r = -0.416, P < 0.05). In the patients with AIHA/Evans syndrome, the quantity of bone marrow CD5+ B lymphocytes showed significantly positive correlation with serum indirect bilirubin level (r = 1.00, P < 0.05). In Evans syndrome patients, the quantity of CD5+ B lymphocytes in bone marrow showed significantly positive correlation with platelet-associated immunoglobulin G (r = 0.761, P < 0.05) and platelet-associated immunoglobulin M ( r = 0.925, P < 0.05). The quantity of CD5+ B lymphocytes in bone marrow of all hemocytopenic patients showed significantly negative correlation with treatment response (tau-b = -0.289, P < 0.05) , but had no correlation with colony forming unit-erythroid (r = -0.205, P > 0.05) or colony forming unit-granulocyte-macrophage colonies (r = -0.214, P > 0.05). CONCLUSIONS: The quantity of bone marrow CD5+ B lymphocytes in the patients with autoimmune hemocytopenia significantly increases and is correlated with disease severity and clinical response, which suggest that CD5+ B lymphocytes might play an important role in the pathogenesis of autoimmune hemocytopenia.

Burden of abnormal hematopoietic clone in patients with myelodysplastic syndromes.

OBJECTIVE: To investigate the role of the burden of abnormal hematopoietic clone in the development of myelodysplastic syndromes (MDS). METHODS: The ratio of the bone marrow cells with abnormal chromosomes to the total counted bone marrow cells was regarded as the index of MDS clone burden. The disease severity related parameters including white blood cell count, hemoglobin, platelet count, lactate dehydrogenase level, bone marrow blast, myeloid differentiation index, micromegakaryocyte, transfusion, interleukin-2, tumor necrosis factor (TNF), CD4+ and CD8+ T cells of MDS patients were assayed, and the correlations between those parameters and MDS clone burden were also analyzed. RESULTS: The clone burden of MDS patients was 67.4% +/- 36.2%. MDS clone burden positively correlated with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin, and erythrocytes in high clone burden (> 50%) and low clone burden ( < or = 50%) groups were 7.78% +/- 5.51% and 3.45% +/- 3.34%, 56.06 +/- 14.28 g/L and 76.40 +/- 24.44 g/L, (1.82 +/- 0.48) x 10(12)/L and (2.32 +/- 0.66) x 10(12)/L, respectively (all P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (0.274 +/- 0.719) x 10(9)/L and (0.455 +/- 0.206) x 10(9)/L, respectively (P < 0.05). CD8+ T lymphocytes of MDS patients and normal controls were (0.240 +/- 0.150) x 10(9)/L and (0.305 +/- 0.145) x 10(9)/L, respectively. The serum level of interleukin-2 of MDS patients (6.29 +/- 3.58 ng/mL) was significantly higher than normal control (3.11 +/- 1.40 ng/mL, P < 0.05). The serum level of TNF of MDS patients and normal control group were 2.42 +/- 1.79 ng/mL and 1.68 +/- 0.69 ng/mL, respectively. The ratio of CD4 to CD8 was higher in high clone burden MDS patients (1.90 +/- 0.52) than that in low clone burden patients (0.97 +/- 0.44, P < 0.05). CONCLUSION: The quantitive clonal karyotype abnormalities and deficient T cell immunity are important parameters for evaluating MDS severity and predicting its progression.

[The prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes].

OBJECTIVE: To investigate the prognostic value of quantitative chromosomal abnormality in myelodysplastic syndromes (MDS). METHODS: Chromosomal karyotypes in seventy-one MDS patients' were analyzed quantitatively. Based on the number of abnormal metaphase in 20 counted metaphases, the patients were divided into three groups: no abnormal karyotypes, abnormal metaphases less than or equal to five, and that more than five. The leukemia transformation rate, death rate and survival time between these three groups were compared. RESULTS: Forty-four cases (62.0%) had abnormal karyotypes. The incidences of abnormal karyotypes in RA, RCMD and RAEB were 76.9%, 55.8% and 75.0%, respectively, being no significant difference (P > 0.05). Among the abnormal karyotypes, complex abnormality with two or more abnormal karyotypes was most common and accounted for 47.7%. The frequencies of trisomy 8, monosomy 7 and del 20q were 18.2%, 4.5% and 4.5%, respectively. Other kinds of abnormal karyotypes totally accounted for 25%. There were 27 cases of group 1, 28 of group 2 and 16 of group 3. Eighteen cases (25.4%) transformed to acute leukemia. The incidences of leukemia transformation in group 1, 2 and 3 were 18.5%, 25% and 37.5%, and the death rates were 29.6%, 42.9% and 56.3%, respectively. The median survival times were 60, 47 and 24 months respectively. CONCLUSION: The quantitative chromosome abnormality has prognostic value in MDS.

[Study on the peripheral blood dendritic cells subtypes and the expression of co-stimulating molecules on dendritic cells and B cells in severe aplastic anemia patients].

OBJECTIVE: To investigate the quantities of monocyte-derived dendritic cell precursors (pDC1) and plasmacytoid dendritic cell precursors (pDC2) in peripheral blood mononuclear cells (PBMC) of severe aplastic anemia (SAA) patients before and after immune suppressive therapy (IST), the ratio of the pDC1 to pDC2, and the expression of co-stimulating molecules (CD80, CD86, CD40) on dendritic cells (DC) and B cells in SAA patients. METHODS: By means of three color monoclonal antibody labeling technology, the quantities and ratio of pDC1 and pDC2 in PBMC were detected in 26 SAA patients at active phase, 13 at recovery phase and 15 normal controls respectively. The aforementioned parameters of 10 SAA patients were tested before and 2 months after IST. The expression of CD80, CD86 and CD40 on DC and B lymphocytes were detected in 16 SAA patients and 15 normal controls. RESULTS: The percentages of pDC1 and the ratio of pDC1/pDC2 of controls were (0.41 +/- 0.05)% and 1.58 +/- 0.18 respectively, and those of SAA patients at active phase were (0.67 +/- 0.13)% and 2.70 +/- 0.32 respectively, [pDC1 (P < 0.05); pDC1/ pDC2 ratio (P < 0.01)]. The aforementioned parameters in convalescent SAA patients decreased to (0.43 +/- 0.10)%, and 1.78 +/- 0.36 respectively, being no difference from those of normal controls. The percentages of pDC1 and pDC2 in 10 SAA patients were (0.87 +/- 0.31)%, and (0.35 +/- 0.09)%, before IST, and (0.24 +/- 0.09)%, (0.14 +/- 0.04)%, after IST, being significantly decreased (P < 0.05). The percentages of CD86 expression on DC of controls was (11.97 +/- 4.31)%, and that of SAA patients was (29.84 +/- 3.02) % (P < 0.05). The percentages of CD80, CD40 and CD86 expression on lymphocytes of controls were (2.57 +/- 0.44)%, (7.34 +/- 1.22)% and (1.86 +/- 1.11)%, respectively, and those of SAA patients were (5.17 +/- 0.68)%, (8.85 +/- 2.94)% and (5.98 +/- 0.96)% respectively (P < 0.05, P < 0.01). The percentage of CD86 expression on B lymphocytes in controls was 8.04 +/- 0.66%, and in SAA patients was (20.46 +/- 2.78)%, (P < 0.05). CONCLUSION: The pDC subtypes were abnormal and the percentage of pDC1 is increased in SAA patients, which are associated with stage of this disease. DC and B Lymphocytes in SAA patients upregulated expression of costimulatory molecules (CD86) which cause the T lymphocyte abnormally activated.

[Study on the burden of abnormal hematopoietic clone of the patients with myelodysplastic syndromes and its implications].

OBJECTIVE: To investigate the abnormal hematopoietic clone burden of the patients with myelodysplastic syndromes (MDS) and its clinical implication. METHODS: The ratio of the metaphase with abnormal karyotypes to the total was regarded as the index of MDS clonal burden. Thirteen parameters were assayed and the correlations between these parameters and MDS clone burden were analysed. RESULTS: The clonal burden of MDS patients was (67.4 +/- 36.2)%. It correlated positively with bone marrow blasts (r = 0.483, P < 0.05), negatively with hemoglobin level (r = -0.445, P < 0.05). The number of blasts, hemoglobin and erythrocytes in high clonal burden (>50%) and low clonal burden (< or = 50%) groups were significantly different (P < 0.05). CD4+ T lymphocytes of MDS patients and normal controls were (274.18 +/-71.85) x 10(6)/L and (454.82 +/- 205.88) x 10(6)/L (P < 0.05) respectively. CD8+ T lymphocytes between MDS patients and normal controls had no difference. The serum level of IL-2 of MDS patients and normal control groups were (6.29 +/- 3.58) g/L and (3.11 +/- 1.40) microg/L (P < 0.05) respectively; but no difference in the serum level of TNF between MDS and control groups. The ratio of CD4+ to CD8+ in high clonal burden patients was 1.90 + 0.52, and in low clonal burden patients was 0.97 +/- 0.44 (P < 0.05). CONCLUSION: The clonal burden and deficient T cell immunity are the indicators for predicting MDS patients clinical progression.

[The response of bone marrow hematopoietic cells to G-CSF in paroxysmal nocturnal hemoglobinuria patients].

OBJECTIVE: To study the response of hematopoietic cells (HSC) to granulocyte colony stimulating factor (G-CSF) in paroxysmal nocturnal hemoglobinuria (PNH) patients. METHODS: (1) Bone marrow mononuclear cells (BMMNC) from 17 PNH patients and 12 normal subjects were inoculated into semisolid culture media containing or not G-CSF (50 ng/ml). The cluster/colony forming unit-granulocyte/monocyte (CFU/cFU-GM) were counted and compared. (2) BMMNC of 20 PNH patients and 12 normal controls were triply stained for CD34, CD59 and G-CSF receptor CD114/stem cell factor receptor (C-KIT) CD117 and assessed by FCM. The CD34(+) cells were identified as CD34(+)/CD59(+) and CD34(+)/CD59(-). Percentage of CD114 and CD117 expression in each cell population was calculated. RESULTS: (1) PNH cFU-GM without G-CSF were (112.41 +/- 22.74)/10(5) BMMNC, while with G-CSF: (133.82 +/- 25.85)/10(5) BMMNC and normal cFU-GM were (190.33 +/- 36.05)/10(5) BMMNC, (309.42 +/- 92.94)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC formed less cFU-GM than control did, both of the two kinds of BMMNC responded to G-CSF well (P < 0.05), but the increment of PNH cFU-GM yields was less than that of the normal control (P < 0.05). CFU-GM yields of PNH BMMNC without G-CSF were (24.29 +/- 9.05)/10(5) BMMNC, with G-CSF were (27.53 +/- 10.65)/10(5) BMMNC, while normal control were (77.42 +/- 36.01)/10(5) BMMNC and (98.00 +/- 43.14)/10(5) BMMNC, respectively. Whether with or without G-CSF, PNH BMMNC showed less CFU-GM yields than that of control (P < 0.05). (2) The percentage of CD114 positive cells in PNH CD34(+)CD59(+) BMMNC was (73.34 +/- 29.40)% and that in PNH CD34(+)CD59(-) BMMNC and in control CD34(+)CD59(+) BMMNC were (32.70 +/- 6.89)% and (58.52 +/- 29.99)%, respectively. The percentage of CD114 expression in PNH CD34(+) CD59(-) BMMNC was less than that in the other two groups (P < 0.05). The percentages of CD117 positivities on the PNH CD34(+)CD59(+) BMMNC were (76.90 +/- 22.08)%, PNH CD34(+) CD59(-) (36.03 +/- 7.69)% and control CD34(+) CD59(+) (80.28 +/- 13.36)%, respectively (P < 0.01). CONCLUSION: In vitro, BMMNC of normal control grow better, and respond better to G-CSF than PNH BMMNC do. PNH CD34(+)CD59(-) BMMNC express less G-CSF receptor and C-KIT than PNH CD34(+)CD59(+) and normal CD34(+)CD59(+) BMMNC do, which may be the reason that abnormal PNH clone grow worse than the normal clones do.

[The clinical features of hepatitis associated aplastic anemia].

OBJECTIVE: To analyse the proportion of hepatitis associated aplastic anemia (HAAA) in severe aplastic anemia (SAA) and its clinical features of HAAA. METHODS: All newly diagnosed SAA cases in our department in the recent 5 years were analyzed. A case-control study was undertaken to investigate the differences of clinical and laboratory features between HAAA and non-hepatitis associated SAA (non-HASAA) patients. RESULTS: The proportion of HAAA in SAA was 3.3%. There was no significant difference in PB cell counts, bone marrow hematopoiesis status and the amount of blood transfusion between HAAA and non-HASAA patients. Sera from 13 patients with HAAA were tested for antibodies to hepatitis viruses A, B, and C and hepatitis B surface antigen. Twelve (92.3%) of them had negative serologic results for the tests and only one (7.7%) had a positive result for HBsAg and HBeAg. Serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were decreased prior to the diagnosis in twelve (92.3%) of the 13 HAAA patients. The percentage of CD4(+) cells in HAAA patients was significantly lower than that in non-HASAA patients (P < 0.05). HAAA patients had higher percentages of CD8(+) cells (P < 0.05) and lower ratios of CD4(+)/CD8(+) (P < 0.05). The early infection rate of the HAAA patients was significantly higher than that of non-HASAA patients (84.6% vs 42.3%, P < 0.05), with different mortalities (61.5% vs 15.4%, P < 0.05). The 2-year survival rate of HAAA patients was significantly lower than that of non-HASAA patients (16.6% vs 83.2%, P < 0.01). CONCLUSION: The proportion of HAAA in SAA was 3.3%. Most of HAAA were associated with non-A, non-B and non-C hepatitis virus. Compared with that of non-HASAA, the abnormality of T cell immunity of HAAA was more severe, with a higher frequency of early infection and a higher mortality rate.

[Apoptosis and proliferation of CD34 positive bone marrow cells in patients with polycythemia vera].

OBJECTIVE: To study the apoptosis and proliferation of CD(34) positive (CD(34)(+)) bone marrow cells (BMC) in patients with polycythemia vera (PV). METHODS: The expression of Annexin V and Ki67 of the CD(34)(+) BMC in 20 PV patients and control cases [10 essential thrombocythemia (ET), 12 normal persons] were assessed by bicolor flow cytometry (FCM), and the correlation between apoptosis and clinical situation was analysed in PV patients. RESULTS: The Annexin V expressions of CD(34)(+) BMC were (15.96 +/- 1.45)% in PV patients and (15.53 +/- 1.76)% in ET patients which were lower than that in normal subjects [(23.61 +/- 3.89)%, (P < 0.05)]. The Ki67 expression of CD(34)(+) BMC was (48.79 +/- 11.68)% in PV patients and (49.60 +/- 9.98)% in ET patients, which were significantly higher than that in normal controls (33.87 +/- 6.82)%. The ratio of apoptosis/proliferation in PV patients was 0.33 +/- 0.10 and in ET patients 0.32 +/- 0.02 which were significantly lower than that in normal controls 0.72 +/- 0.11 (P < 0.01). The apoptosis of CD(34)(+) BMC was negatively correlated with the hemoglobin (Hb) levels (r = -0.481, P = 0.037), white blood cells (WBC) (r = -0.538, P = 0.026) and the numbers of endogenous erythroid colony (EEC) (r = -0.632, P = 0.50), and the ratio of apoptosis/proliferation was negatively correlated with the Hb (r = -0.537, P = 0.018) and WBC (r = -0.667, P = 0.003) in PV patients. CONCLUSION: There were lower apoptosis and higher proliferation in CD(34)(+) BMC of PV patients. Lower apoptosis was correlated with the severity of the disease.

[Preliminary study of DA or HA regimen chemotherapy for the treatment of refractory and relapsed paroxysmal nocturnal hemoglobinuria].

OBJECTIVE: To observe the efficacy and side effect of DA/HA regimen chemotherapy for the treatment of refractory and relapsed paroxysmal nocturnal hemoglobinuria (PNH). METHODS: Eight patients with refractory and relapsed PNH were treated with DA/HA regimen chemotherapy. Three patients were treated with DA (DNR 40 mg/d, i.v.drip, the first and the second day; 20 mg/d, i.v.drip, the third day; Ara-C 100 mg/d, i.v.drip, for 5 days) and 5 patients with HA (HHT 2 - 3 mg/d, i.v.drip, for 5 days; Ara-C 100 mg/d, i.v.drip, for 5 days). RESULTS: All the 8 patients responded well: the PNH clone was diminished in five patients. Hemolysis was remitted in 6 cases. Five patients showed improvement in hematological parameters. The dosage of corticosteroid was decreased in all of them. No serious side effect was revealed. CONCLUSION: DA/HA regimen chemotherapy was safe and effective for refractory and relapsed PNH patients.

Endogenous erythroid colony assay in patients with polycythemia vera and its clinical significance.

BACKGROUND: Polycythemia vera (PV) is a malignant disorder of hemaopoietic stem cells which is characterized by clonal hyperproliferation and a low rate of apoptosis. This study was to assess endogenous erythroid colony (EEC) formation in the bone marrow of PV patients and determine its clinical significance. METHODS: The bone marrow mononuclear cells of 26 patients with PV, 2 patients with secondary erythrocytosis (SE), and 19 normal controls were cultured by Marsh's method for EEC evaluation, and the clinical significance was evaluated. RESULTS: EECs appeared in 25 patients with PV but not in 2 patients with SE and 19 normal controls. The number of EECs and the EEC ratio [EEC/erythropoietin (EPO)-dependent colony forming unit-erythroid (CFU-E)] in PV patients positively correlated with hemoglobin (Hb) levels. Their EEC number did not correlate with white blood cell (WBC) counts, platelet (PLT) counts, or leukocyte alkaline phosphatase (LAP) scores. Their EEC did not correlate with serum EPO levels. Fifteen patients with PV were treated with hydroxyurea (Hu) and/or interferon-alpha (IFN-alpha). Their EEC ratio before treatment positively correlated with the treatment time required for complete remission (CR) and negatively correlated with the time before relapse. The EEC numbers of 7 PV patients treated with Hu/IFN-alpha decreased after the blood cell counts dropped to normal levels. There was a positive correlation between the EEC ratio and the incidence of attacks of vascular thrombosis in PV patients. The numbers of apoptosised bone marrow mononuclear cells in PV patients were lower than those in normal controls. The EEC numbers of PV patients negatively correlated with the rate of apoptosis of bone marrow mononuclear cells. CONCLUSIONS: EEC formation is characteristic in PV patients. EEC number in PV patients positively correlates with Hb levels, the time required for CR, and the incidence of attacks of vascular thrombosis. EEC number negatively correlates with the time before relapse. Bone marrow suppressive treatment might decrease EEC number. Thus, EEC number is a sensitive and specific parameter reflecting the abnormal hematopoietic clone burden induced by polycythemia vera. EEC number is an important diagnostic parameter for PV patients.

[Expression of apoptosis related proteins in CD34 positive bone marrow cells of patients with polycythemia vera].

OBJECTIVE: To investigate the expression of apoptosis receptor FAS (CD95) and apoptosis related protein Bcl-2 and Bax in CD34 positive bone marrow cells of the patients with polycythemia vera (PV). METHODS: The expressions of apoptosis receptor FAS (CD95) and apoptosis related protein Bcl-2 and Bax in bone marrow CD34(+) cells from 21 PV patients, 8 essential thrombocythemia (ET) and 11 normal persons were assessed by bicolor flow cytometry (FCM), and the expressions of Bcl-2 and Bax mRNA were assessed by RT-PCR, and their correlation was analysed. RESULTS: There was no difference between the expressions of CD95 in CD34(+) bone marrow cells of PV patients (42.65 +/- 15.56)%, and that of ET patients (45.31 +/- 17.62)% and of normal person (37.55 +/- 15.19)% (P > 0.05). There was no difference between the expression of Bax in CD34(+) bone marrow cells of PV patients (35.83 +/- 9.33)% and of normal persons (41.65 +/- 9.04)% (P > 0.05). The expression of Bcl-2 in CD34(+) bone marrow cells of PV patients (79.35 +/- 14.43)% was significantly higher than that of normal controls (55.84 +/- 13.43)% (P < 0.01). The ratio of Bax/Bcl-2 of PV patients (0.47 +/- 0.14) was significantly lower than that in normal controls (0.76 +/- 0.24) (P < 0.01). The expression of Bcl-2 mRNA in PV patients' bone marrow hematopoietic cells was higher than that of normal controls (P < 0.01). There was no difference between the expression of Bax mRNA in bone marrow hematopoietic cells of PV patients and that of normal controls. Bcl-2 expression was negatively correlated with Annexin V expression in CD34(+) bone marrow cells of PV patients. CONCLUSION: Over-expression of Bcl-2, one of anti-apoptosis genes, in CD34(+) bone marrow cells might be involved in the lower apoptosis of PV patients.

[The clinical features of severe aplastic anemia patients with complication of infection].

OBJECTIVE: To study the clinical features of severe aplastic anemia (SAA) patients with complication of infection. METHODS: A retrospective analysis of prevalence of infection occurring in 229 SAA patients, their bacterial spectrum, and the effect of GM-CSF or G-CSF on the infection were done. RESULT: The prevalence of infection in SAA patients was 86.0%, among which 54.2% was infected with gram-positive organisms, 40.0% with gram-negative bacilli and 5.8% with fungal infections. Septicemia occurred mostly with E. coli and Pseudomonas infection. Patient's neutropenia was significantly related to the infection. The patients with neutrophil count less than 0.2 x 10(9)/L had more frequent and severe infection. Age, hemoglobin level, subtype of T lymphocytes and antithymocyte globulin therapy were not related to infection. Prophylaxis usage of floxacin could not reduce patient' gastrointestinal infection. The total mortality of SAA patients with infection was 23.1%. Pulmonary infection and septicemia increased mortality, and GM-CSF/G-CSF therapy reduce mortality. CONCLUSION: SAA patients were at high risk of infection which was significantly associated with severe neutropenia. GM-CSF or G-CSF therapy exerts an assistant role to antibiotics in controlling the infections.

[An analysis of relapse and risk factors of autoimmune hemolytic anemia and Evans syndrome].

OBJECTIVE: To analyse the relapse rate and risk factors of autoimmune hemolytic anemia (AIHA) and Evans syndrome. METHODS: Fifty two cases of AIHA and Evans syndrome in remission being followed up for 1 - 14 years (median time 3.8 years) were analysed for relapse rate. The risk factors of relapse were analysed by case-control study. RESULTS: The total relapse rate of these AIHA and Evans syndrome patients was 57.7%, and the median remission duration to the first relapse was 9 months. The relapse rates in patients with negative Coombs test, warm autoantibodies and both of warm and cold autoantibodies were 30.8% (4/13), 54.0% (13/24) and 86.7% (13/15), respectively. The relapse rate in patients with cold antibody was the highest (P < 0.05). The relapse rate in patients with antibody titer >or= 100 was 92.9% (13/14) and was higher than that in patients with antibody titer < 100 [59.5% (13/22)] (P < 0.05). Patients treated with prednisone and cyclosporin relapsed less than those treated with prednisone alone, and the relapse was related to the therapy course of prednisone and CsA. CONCLUSION: Because of the high relapse rate, AIHA and Evans syndrome should be treated according to the class of autoantibodies, and with longer course of prednisone and cyclosporin and prophylaxis of infection.

[Detection for endogenous erythroid colony in the patients with polycythemia vera and its clinical significance].

OBJECTIVE: To investigate the growth of endogenous erythroid colony (EEC) in polycythemia vera (PV) patients and its clinical significance. METHODS: Bone marrow mononuclear cells of 26 PV patients, 2 secondary erythrocytosis (SE) and 19 normal controls were cultured by Marsh's method for EEC. RESULTS: 1. EEC was present in 25/26 (96.2%) PV patients and was not found in 2 SE patients and 19 normal controls. 2. The number of EEC and the ratio of EEC/Epo-dependent CFU-E (EEC ratio) were positively correlated with the hemoglobin (Hb) levels (r = 0.608, P = 0.01) in PV patients, but did not correlate with white blood cell (WBC) counts, platelet counts and neutrophil alkaline phosphatase scores. 3. EEC did not correlate with PV patients' serum Epo levels (r = 0.518, P = 0.125). 4. Fifteen PV patients were treated with hydroxyurea and/or interferon-alpha. Their EEC ratio before treatment was correlated positively with the time required for complete remission (CR) (r = 0.651, P = 0.009) and negatively with the time before relapsing (r = -0.529, P < 0.02). 5. EECs of 7 PV patients treated with HU/IFN were decreased after their blood cell counts normalization. 6. There was a positive correlation between the EEC ratio and the attacks of vascular thrombosis (r = 0.524, P = 0.01). (7) The apoptosis of bone marrow mononuclear cells of PV patients was less than that of normal controls. PV patients' EEC was negatively correlated with the apoptosis of their bone marrow mononuclear cells (r = -0.192, P < 0.045). CONCLUSION: EEC is peculiarly present in PV patients, and is a sensitive parameter in reflecting the abnormal hematopoietic clone burden and in diagnosing and monitoring the disease.