Aged ; CD59 Antigens ; immunology ; Female ; Granulocytes ; immunology ; Humans ; Myelodysplastic Syndromes ; immunology ; pathology

The myelodysplastic syndromes (MDS) are a group of heterogeneous hematological disorders and characterized by abnormalities of bone marrow myeloblasts in morphology and count, ineffective hematopoiesis, decrease of peripheral blood cells and high risk of transforming into acute myeloid leukemia. In this review, the morphological examination of bone marrow, bone marrow biopsy, karyotyping analysis of chromosome, and the significance of flow cytometric immunophenotyping for diagnosis and prognosis of MDS patients are discussed.
Bone Marrow ; pathology ; Humans ; Immunophenotyping ; Karyotyping ; Myelodysplastic Syndromes ; genetics ; immunology ; pathology

This study was aimed to evaluate whether mesenchymal stem cells (MSCs) obtained from patients with myelodysplastic syndrome possess immunosuppressive effect. MSCs from bone marrow samples of MDS patients were isolated, cultured and expanded. MSCs were morphologically analyzed and their immunophenotype were determined by flow cytometry. Various amounts of MSCs were added into one-way mixed lymphocyte reaction. MSCs from MDS patients were tested for their ability to suppress in vitro proliferation of autologous and allogeneic peripheral blood lymphocytes (PBLs). The results showed that 3 x 10(3 - 1) x 10(5) MSCs from MDS patients could inhibit autologuous PBLs proliferation to (66.9 +/- 20.1)% - (30.2 +/- 5.9)% of maximal response, as well as inhibit allogeneic PBLs proliferation to (56.6 +/- 14.7)% - (20.5% +/- 9.7)% of maximal response, as compared with inhibitory ability of MSCs from healthy donors, there was no significant difference (P>0.05). It is concluded MSCs from patients with myelodysplastic syndrome also possess immunosuppressive activity.
Bone Marrow Cells ; immunology ; pathology ; Cell Proliferation ; Cells, Cultured ; Humans ; Immune Tolerance ; Lymphocyte Culture Test, Mixed ; Lymphocytes ; cytology ; Mesenchymal Stromal Cells ; immunology ; pathology ; Myelodysplastic Syndromes ; immunology ; pathology

The study was aimed to compare the effects of T-cell suppression mediated by mesenchymal stem cells (MSC) from normal individuals and myelodysplastic syndromes (MDS) patients. MSC were cultured from the bone marrow of 12 healthy volunteers and 12 MDS patients, the morphology, surface markers and expression of several cytokines of MSC from normal individuals and MDS patients were compared, and the effects of T-cell suppression were tested in the following assays: phytohemaglutinin (PHA)-primed cultures, mixed lymphocyte reaction (MLR), cell cycle of T-cell after PHA-primed cultures and apoptosis of T-cell as well. The results showed that the MSC from normal individuals and MDS patients were similar in morphology, proliferation and surface markers. The suppressions of T-cell proliferation induced by PHA and alloantigens mediated by MDS-MSC were significantly lower than that of normal MSC. More T-cells were arrested in G0/G1 phase by normal MSC, while the effects were deficient by MDS-MSC. The suppression of T-cell activation mediated by MDS-MSC was also lower than that of normal MSC, but suppression effect on T-cell apoptosis increased. The cytokines TGF-beta1, 3, FasL expressed by MDS-MSC were reduced as compared with normal MSC, but TGF-beta2 expression increased in MDS-MSC. It is concluded that although the morphology, proliferation and cell surface markers of MDS-MSC are normal, the T-cell suppression mediated by MDS-MSC is deficient as compared with normal controls. Whether these abnormalities are relevant to the pathogenesis of aplastic anemia remains to be determined.
Adult ; Aged ; Bone Marrow Cells ; cytology ; physiology ; Female ; Humans ; Immune Tolerance ; immunology ; physiology ; Male ; Mesenchymal Stromal Cells ; immunology ; physiology ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology ; T-Lymphocytes ; immunology

Although it has been shown that the percentage of bone marrow blasts in myelodysplastic syndrome (MDS) constitute the only independent determinant of survival and progression to acute leukemia, the great variability in survival among patients with MDS of similar percentage of blasts has prompted us to investigate new objective, independent prognostic parameters for the selection of high-risk patients. It was suggested that CD34 antigen expression adversely affected the prognosis of acute myelogenous leukemia. However, no study has been published so far on clinical and prognostic significance of CD34 antigen expression in MDS. Bone marrow biopsies from 58 patients diagnosed as primary MDS were studied using QBEND/10, a monoclonal antibody which recognized the human progenitor CD34 antigen on routine aldehyde-fixed, paraffin-embedded samples. The high percentage of CD34-positive cells (above 3% of total bone marrow nucleated cells) was predominantly observed in cases with RAEB-T, CMML, and to a lesser degree in RAEB. But neither age, hemograms, bone marrow findings including percentage of blasts, ALIP, nor leukemic transformation correlated with the percentage of CD34-positive cells. The median actuarial survival time in the high positive group was significantly shorter (12.0 months) than that of the low group (30.0 months; p = 0.028). The high CD34 aggregate (> or = 3) was selectively found in cases with RAEB, RAEB-T, and CMML. The percentage of bone marrow blasts (p = 0.007) and ALIP (p = 0.030) significantly correlated with number of CD34 aggregates.
Adult ; Aged ; Antigens, CD/*analysis ; Antigens, CD34 ; Biopsy ; Bone Marrow/*immunology/*pathology ; Human ; Immunohistochemistry/methods ; Middle Age ; Myelodysplastic Syndromes/*immunology/*pathology ; Staining and Labeling ; Tumor Markers, Biological

OBJECTIVETo investigate the abnormal hematopoietic clone burden of the patients with myelodysplastic syndromes (MDS) and its clinical implication.

METHODSThe 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.

RESULTSThe 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).

CONCLUSIONThe clonal burden and deficient T cell immunity are the indicators for predicting MDS patients clinical progression.

Adolescent ; Adult ; Aged ; Bone Marrow Cells ; pathology ; Chromosome Aberrations ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; pathology ; T-Lymphocytes ; immunology

This study was aimed at exploring the immunophenotypic features of blasts in patients with myelodysplastic syndromes (MDS). Four-color flow cytometry using conventional and secondary gating strategies was used to immunophenotype blasts and the CD34 positive cells in bone marrow nucleated cells of 29 patients with MDS. The results showed: (1) with progression of MDS from RA/RAS, RAEB to RAEB-T, the proportion of CD34(+) cells were gradually increased from 8.0%, 46.4% to 76.8% (P < 0.05); (2) using CD45 vs SSC gating strategy, with the transformation of RA/RAS, RAEB to RAEB-T, the expression of HLA-DR, CD13, CD33, CD117 were also gradually increased (P < 0.05), and the expression of CD15 was gradually decreased (P < 0.05); (3) using CD45 vs CD34 gating strategy, the expression of HLA-DR, CD13, CD33, CD117 on blasts were higher by secondary gating method than those by conventional gating (P < 0.05). However, there were no significant difference (P > 0.05) in the expression of above-mentioned antigens on CD34(+) cells among different MDS subtypes. It is concluded that conventional gating method can reflect MDS progression from RA/RAS, RAEB to RAEB-T, and secondary gating strategy may accurately reflect the biological features of blasts in MDS. Abnormal expression of CD34 is related to the immaturity level and heterogeneity of blast cells, which is beneficial to the diagnosis of clinically suspected MDS incapable of diagnosing with morphology and cytogenetics.
Adult ; Aged ; Antigens, CD34 ; analysis ; Bone Marrow Cells ; immunology ; pathology ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology

This study was purposed to analyse the characteristics of blast immunophenotype in patients with refractory anemia with excess blasts, type 1 (RAEB-1) and refractory anemia with excess blasts, type 2 (RAEB-2) by flow cytometry and investigate its diagnostic significance, as well as to compare the blast rate detected by FCM and bone marrow smear (BMS). FCM was used to count the blast rate and detect the expression of its antigens in 29 cases of MDS. The result indicated that: (1) The rate of the blasts detected by FCM and BMS was not statistically significant between patients with RAEB-1 and with RAEB-2 (P > 0.05); (2) Out of 13 patients with RAEB-1, the blasts of 10 cases, 12 cases, 8 cases, 11 cases, 11 cases, 3 cases, 7 cases, 0 case, 0 case, 3 cases and 2 cases patients had positive expressions of CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, separately. The blasts of 12 cases, 13 cases, 3 cases, 12 cases, 15 cases, 7 cases, 5 cases, 0 case, 1 case, 4 cases and 2 cases among 16 patients with RAEB-2 positively expressed CD34, HLA-DR, CD117, CD33, CD13, CD15, CD11b, CD3, CD19, CD7 and CD56, respectively. However, there was no significant difference in the expression of antigens in blasts of the bone marrow between the patients with RAEB-1 and with RAEB-2 (P > 0.05); (3) The positive expression for CD15 and CD11b in blasts was found in 10 cases and 12 cases, respectively; (4) The positive expression for CD19, CD7 and CD56 was observed in 1, 7 and 4 cases, respectively. None of the 29 cases of MDS was positive for CD3. It is concluded that FCM can reveal the characteristics of immunophenotypic abnormalities of the blasts in patients with MDS and provide the important information for diagnosis and differential diagnosis of MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; pathology ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; diagnosis ; immunology ; pathology ; Prognosis ; Young Adult

The study was aimed to investigate the abnormality of immunophenotypes in patients with myelodysplastic syndrome (MDS) and its role in the identification of MDS. The cell immunophenotypes of 136 patients with hypocytosis accompanied by abnormal hematopoiesis of bone marrow were detected by flow cytometry, the detected results were evaluated by flow cytometric scoring system (FCSS), and the sensitivity and specificity of positive results were determined by FCSS also. The correlation of results detected by FCSS to traditional diagnosis method was analysed. The results indicated that 111 out of 136 cases were diagnosed as MDS, and 25 were diagnosed as non-MDS. Among 111 MDS cases, 85 cases were FCSS positive, 18 cases were FCSS intermediate and 8 cases were FCSS negative, whereas in 25 non-MDS cases 24 cases were FCSS negative, 1 case was FCSS intermediate and no case was FCSS positive. The sensitivity of FCSS in identification of MDS was 76.6%, and the specificity of FCSS was 100%. There was a good correlation of FCSS to traditional method (R = 0.613, p = 0.000). It is concluded that the various abnormalities of immunophenotyping are found in patients with MDS, in which the main immunophenotype abnormality and the abnormality involving two cell lineages are key points to distinguish MDS from non-MDS.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; pathology ; Female ; Flow Cytometry ; methods ; Humans ; Immunophenotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; pathology ; Sensitivity and Specificity ; Young Adult

OBJECTIVETo explore polarization of T lymphocyte and its relationship with apoptosis of marrow cells in patients with myelodysplastic syndromes (MDS).

METHODSMeasurements of Th1, Th2, Tc1, Tc2 subsets in bone marrows from 34 patients with MDS and 13 normal controls were performed by flow cytometry. INF-gamma and TNF-alpha in marrow serum were determined by ELISA (Enzyme-linked immunosorbent assay). Apoptosis index of marrow cells was detected by TUNEL (TdT-mediated dUTP nick end labeling). Correlations between Th1, Th2, Tc1, Tc2 subsets and INF-gamma, TNF-alpha levels as well as apoptosis index were analyzed, and relationship between TNF-alpha, INF-gamma levels and apoptosis index was also investigated.

RESULTS(1) The percentage of Th1 cells [(10.1 +/- 1.6)%], Tc1 cells [(24.0 +/- 3.6)%] and Tc1/Tc2 ratio (50.0 +/- 11.1) was significantly increased in patients with MDS than in normal controls [(4.0 +/- 0.5)%, (5.8 +/- 0.6)% and 13.4 +/- 2.7, respectively]. Levels of INF-gamma [(58.6 +/- 21.7) microg/L] and TNF-alpha [(15.7 +/- 3.8) microg/L] in marrow serum of MDS patients was markedly elevated compared to normal controls [0 and (0.3 +/- 0.2) microg/L, respectively]. An increased apoptosis index of nucleated cells was observed in MDS patients [(7.8 +/- 1.5)%] as compared to controls [(2.1 +/- 0.3)%, P < 0.05]. The Th1 cell percentage showed a positive correlation with the levels of INF-gamma and TNF-alpha (r = 0.38, P < 0.05 and r = 0.39, P < 0.05, respectively), and with apoptotic index of nucleated marrow cells in MDS patients (r = 0.33, P < 0.05). Furthermore, a positive correlation was observed between INF-gamma, TNF-alpha levels and apoptotic index of marrow cells (r = 0.74, P < 0.01 and r = 0.73, P < 0.01, respectively). (2) Th1, Tc1 cells and Tc1/Tc2 ratio in MDS-RCMD patients was markedly elevated (P < 0.01) but did not in RCMD-RS, RAEB-1 and RAEB-2 patients as compared to normal controls. (3) An elevation in the percentages of Th1, Tcl and Tc1/ Tc2 ratio was detected in patients with IPSS lower-risk but did not in higher-risk group as compared to controls. (4) Increased Th1 and Tc1 percentages and Th1/Th2 and Tc1/Tc2 ratios were observed in RCMD patients with normal karyotype, but did not in those with abnormal karyotype. Conclusions Th1/Th2 and Tc1/Tc2 in bone marrow of MDS patients were unbalanced, polarizing to type I reaction especially in patients with RCMD subtype, IPSS lower-risk and normal karyotype. The increased Th1 cells in bone marrow may account for the increased apoptosis of nucleated marrow cells in MDS, through proapoptotic cytokines such as INF-gamma and TNF-alpha.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Apoptosis ; immunology ; Bone Marrow ; immunology ; metabolism ; pathology ; Female ; Hematopoietic System ; immunology ; Humans ; Interferon-gamma ; metabolism ; Male ; Middle Aged ; Myelodysplastic Syndromes ; immunology ; T-Lymphocytes ; immunology ; Tumor Necrosis Factor-alpha ; metabolism