1.The mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test.
Lei HUANG ; Rong FU ; Li-juan LI ; Hui LIU ; Yi-hao WANG ; Hong-lei WANG ; Tian ZHANG ; Kai DING ; Shao-xue DING ; Er-bao RUAN ; Wen QU ; Jing GUAN ; Guo-jin WANG ; Jia SONG ; Hua-quan WANG ; Yu-hong WU ; Li-min XING ; Hong LIU ; Xiao-ming WANG ; Zong-hong SHAO
Chinese Journal of Hematology 2013;34(5):430-434
OBJECTIVETo investigate the mechanisms underlying bone marrow damage by iron overload in pancytopenic patients with positive BMMNC-Coombs test (IRP).
METHODSTwenty-one iron overloading, 26 non-iron overloading IRP patients and 10 normal controls were enrolled in this study. The expressions of ROS, Bcl-2, Caspase-3 and apoptosis of BMMNC were analyzed by flow cytometry (FCM). Antioxidants were added to iron overloading IRP BMMNC, and then the changes of indices above were detected by FCM. The number and apoptosis of T lymphocytes of IRP patients were also detected.
RESULTSROS and apoptosis of BMMNC, myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones and normal controls (P < 0.05). The expressions of Bcl-2 on BMMNC, erythrocytes and stem cells of iron overloading IRP patients were significantly lower than those of non-iron overloading IRP ones (P < 0.05). The levels of Caspase-3 on myelocytes, erythrocytes and stem cells of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones and normal controls (P < 0.05). After treatment with antioxidants, the expressions of ROS, Caspase-3 and apoptosis of iron overloading IRP BMMNC significantly decreased, but opposite for Bcl-2. The percentages of CD4(+) lymphocytes [ ( 40.86 ± 8.74)%] and CD4(+)/CD8(+) (1.44 ± 0.36) in PB of iron overloading IRP patients were significantly higher than that of non-iron overloading IRP ones [(35.96 ± 7.03)% and 1.14 ± 0.37] and normal controls [(28.00 ± 6.73)% and 0.79 ± 0.21], respectively (P < 0.05), as opposite for CD8(+) lymphocytes (P < 0.05). The apoptosis of CD8(+) lymphocytes [(27.35 ± 10.76)%] and the ratio of CD8(+) apoptosis/CD4(+) apoptosis (2.51 ± 0.81) in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP ones [(15.47 ± 8.99)%] and normal controls (1.39 ± 0.47), respectively (P < 0.05). The apoptosis of erythrocytes and stem cells coated with auto-antibodies in BM of iron overloading IRP patients were significantly higher than those of non-iron overloading IRP and normal controls.
CONCLUSIONMechanisms underlying bone marrow damage by iron overload might be through the follows: ①The increased ROS induced by excessive iron deposition affected the expressions of Caspase-3 and Bcl-2, which caused more BMMNC apoptosis; ②The abnormal number and ratio of T lymphocytes caused by iron overload aggravated the abnormality of immunity of IRP; ③Iron overload may increase the damage to erythrocytes and stem cells coated with auto-antibodies.
Adolescent ; Adult ; Aged ; Bone Marrow ; pathology ; Case-Control Studies ; Caspase 3 ; metabolism ; Coombs Test ; Female ; Humans ; Iron Overload ; Male ; Middle Aged ; Pancytopenia ; immunology ; pathology ; physiopathology ; Proto-Oncogene Proteins c-bcl-2 ; metabolism ; Reactive Oxygen Species ; metabolism ; Young Adult
2.EPOR and TPOR expressions on CD34+ CD59- and CD34+ CD59+ bone marrow cells from patients with paroxysmal nocturnal hemoglobinuria.
Dan WANG ; Rong FU ; Er-Bao RUAN ; Wen QU ; Yong LIANG ; Hua-Quan WANG ; Jun WANG ; Li-Juan LI ; Hui LIU ; Hong-Lei WANG ; Tian ZHANG ; Hong LIU ; Yu-Hong WU ; Li-Min XING ; Guo-Jin WANG ; Xiao-Ming WANG ; Jia SONG ; Jing GUAN ; Zong-Hong SHA
Chinese Journal of Hematology 2011;32(8):543-547
OBJECTIVETo study the expressions of erythropoietin receptor (EPOR) and thrombopoietin receptor (TPOR) on CD34+ CD59- and CD34+ CD59+ bone marrow (BM) cells from patients with paroxysmal nocturnal hemoglobinuria (PNH).
METHODS(1) The expressions of EPOR and TPOR on CD34+ CD59- and CD34+ CD59- BM cells from 26 PNH patients and 16 normal controls were examined by flow cytometry (FCM). (2) The mRNA expression of the EPOR and the TPOR in BM mononuclear cells (BMMNC) from 25 PNH patients and 13 normal controls were examined by RT-PCR.
RESULTS(1) The percentage of EPOR positive cells in PNH CD34+ CD59+ BMMNC [(30.67 +/- 18.30)%] was significantly higher than that in PNH CD34+ CD59- BMMNC [(8.05 +/- 3.51)%] (P < 0.01) and than that in control CD34+ CD59+ BMMNC [(8.24 +/- 6.51)%] (P < 0.01), but there was no obvious difference between the CD34+ CD59-BMMNC in PNH and CD34+ CD59+ BMMNC in control. (2) The percentage of TPOR positive cells in PNH CD34+ CD59+ BMMNC [(28.15 +/- 17.75)%] was significantly higher than that in PNH CD34+ CD59-BMMNC [(15.65 +/- 14.45)%] (P < 0.05) and than that in control CD34+ CD59+ BMMNC [(10.77 +/- .39)%] (P < 0.01), but there was no obvious difference between the CD34+ CD59- BMMNC in PNH and CD34+ CD59+ BMMNC in control. (3) There was no statistic difference in EPOR mRNA and TPOR mRNA expressions in BMMNCs between PNH patients group [(0.41 +/- 0.37) and (0.32 +/- 0.19), respectively] and control group [(0.47 +/- 0.33) and (0.40 +/- 0.29), respectively].
CONCLUSIONThe expression of EPOR and TPOR of PNH patients on BM CD34+ CD59+ cells are significantly higher than those on BM CD34+ CD59- cells. The difference may be due to abnormal transcription of both receptor coding genes.
Adult ; Bone Marrow Cells ; metabolism ; CD59 Antigens ; metabolism ; Case-Control Studies ; Cells, Cultured ; Female ; Flow Cytometry ; Hemoglobinuria, Paroxysmal ; metabolism ; Humans ; Male ; Middle Aged ; Receptors, Erythropoietin ; metabolism ; Receptors, Thrombopoietin ; metabolism ; Young Adult
3.Effect of CD8+ effector T cells on the hematopoiesis pathway damage in the patients with severe aplastic anemia.
Le FENG ; Rong FU ; Hua-quan WANG ; Jun WANG ; Chun-yan LIU ; Li-juan LI ; Hui LIU ; Hong-lei WANG ; Tian ZHANG ; Er-bao RUAN ; Yong LIANG ; Wen QU ; Guo-jin WANG ; Yu-hong WU ; Hong LIU ; Xiao-ming WANG ; Jia SONG ; Jing GUAN ; Li-min XING ; Zong-hong SHAO
Chinese Journal of Hematology 2011;32(9):597-601
OBJECTIVETo investigate the quantity and the pathway to damage hematopoietic cells of CD8+CD25+ and CD8+ HLA-DR+ effector T cells in peripheral blood (PB) of severe aplastic anemia(SAA) patients and explore the immunopathogenesis of SAA.
METHODSThe quantity of CD8+ CD25+ and CD8+ HLA-DR+ cells in PB and the expressions of perforin, granzyme B, tumor necrosis factor-beta (TNF-beta) and FasL in 29 SAA (14 untreated and 15 recovered) patients and 12 normal controls were analyzed by flow cytometry.
RESULTSThe fraction of CD8+ CD25+ T cells in CD8+ T cells was (3.67 +/- 2.58)% in untreated SAA patients, (5.19 +/- 4. 29)% in recovered patients and (4.84 +/- 2.31)% in normal controls, and that of CD8+ CD25+ T cells in CD3+ cells in the three groups was (2.25 +/- 1.35)%, (2.98 +/- 1.35)% and (2.11 +/- 1.88)%, respectively. They had no statistic difference among the 3 groups (P >0.05). The fraction of CD8+ HLA-DR+ T cells in CD8+ T cells was (39.30 +/- 8.13)% in untreated patients, which was significantly higher than that in recovered patients [(20.65 +/- 5.38)%] and controls [(18.34 +/- 6.68)%] (P<0.001), while there was no statistic difference between the latter two groups (P>0.05). CD8+ HLA-DR+ T cells in CD3+ cells was (27.81 +/- 7.10)% in untreated group, which was significantly higher than that of recovered group [(12.02 +/- 3.03)%] and controls [(8.50 +/-2.33)%] (P<0.01). And that in recovered group was higher than that in control group (P<0.05). The expressions of perforin, granzyme B, TNF-beta and FasL of CD8+ HLA-DR+ T cells in untreated group were 8.51%, 96.08%, 72.11% and 94.25% respectively, which were higher than those in recovered group (1.78%, 85.20%, 34.38% and 51.20%) and controls (1.86%, 82.09% ,17.92% and 32.91%). There was no statistic difference between recovered patients and controls (P>0.05).
CONCLUSIONThere were elevated quantity of CD8+ HLA-DR+ T cells and high expressions of perforin, granzyme B, TNF-beta and FasL in SAA, which might contribute to the bone marrow failure.
Adolescent ; Adult ; Anemia, Aplastic ; blood ; metabolism ; pathology ; CD8-Positive T-Lymphocytes ; cytology ; Case-Control Studies ; Child ; Fas Ligand Protein ; metabolism ; Female ; Granzymes ; metabolism ; Humans ; Lymphocyte Count ; Lymphotoxin-alpha ; metabolism ; Male ; Middle Aged ; Perforin ; metabolism ; Young Adult
4.Clinical analysis of 102 blood disease patients with gram positive cocci infection treated with Linezolid.
Jing GUAN ; Rong FU ; Er-Bao RUAN ; Yong LIANG ; Wen QU ; Guo-Jin WANG ; Xiao-Ming WANG ; Hong LIU ; Yu-Hong WU ; Jia SONG ; Hua-Quan WANG ; Li-Min XING ; Zong-Hong SHAO
Chinese Journal of Hematology 2010;31(8):527-530
OBJECTIVETo observe the efficacy and safety of linezolid for the treatment of gram positive coccus infections in hematological disease patients.
METHODSOne hundred and two hematological disease patients with suspected or proven gram positive coccus bacteria infection were enrolled in this study. Linezolid was given at a dosage of 600 mg, iv, q12h. The mean treatment period was (10.82 ± 5.12) days (1 to 51 days) with 74.5% over 7 d and 51.0% over 10 d.
RESULTSAmong 102 patients, 57 were male, 45 female aged 11 to 81 years, with a mean of (45.26 ± 19.15) years. Ninety four cases were nosocomial infection (92.2%) and 8 community infection (7.8%); There were pneumonia in 80 (78.4%), septicemia in 11 (10.8%), and infection of other organsin 11 (10.8%); Forty five cases were proven gram positive coccus bacteria infection, and 57 were suspected infection; Fifty one bacteria strains were isolated from cultivated samples of proven patients, in which 22 were staphylococcus aureus with 19 methicillin resistant 13 hemolytic streptococcus, 9 staphylococcus epidermidis with 7 methicillin resistant 6 enterococcus faecom, and 1 enterococcus hirae. Seven cases were mixed with one kind gram negative bacillus infection, 4 mixed with two kinds of gram negative bacillus infection, and 12 mixed with fungal infection; Total clinical response rates by ITT (intention to treatment) analysis was 69.6%, in which 40 (39.2%) were curative and 31 (30.4%) obviously effective; PP (per-protocol) analysis was 70.9%, in which 39 (41.9%) were curative and 27 (29.0%) obviously effective. Bacteria clearance rate was 70.6%, and in this group the clinical effective rate was 88.9%; Adverse effect rate was 2.9%, being transient thrombocytopenia and increased transaminase.
CONCLUSIONLinezolid is a safe and effective antibiotic used in hematological disease patients complicated with infections of gram positive coccus.
Acetamides ; Anti-Bacterial Agents ; therapeutic use ; Cross Infection ; microbiology ; Gram-Positive Bacterial Infections ; drug therapy ; Gram-Positive Cocci ; Hematologic Diseases ; drug therapy ; Humans ; Linezolid ; Oxazolidinones ; Staphylococcus aureus
5.STAT5 phosphorylation in CD34(+)CD38(-)CD123(+) bone marrow cells of the patients with myelodysplastic syndrome.
Bing-nan LIU ; Rong FU ; Hua-quan WANG ; Li-juan LI ; Lan-zhu YUE ; Er-bao RUAN ; Wen QU ; Yong LIANG ; Guo-jin WANG ; Xiao-ming WANG ; Hong LIU ; Yu-hong WU ; Jia SONG ; Li-min XING ; Jing GUAN ; Jun WANG ; Zong-hong SHAO
Chinese Journal of Hematology 2012;33(6):480-483
OBJECTIVETo investigate the expressions of STAT5 phosphorylation in CD34(+)CD38(-)CD123(+) bone marrow cells of the patients with myelodysplastic syndromes (MDS), and then evaluate the level of activation of STAT5 associated with cell proliferation in MDS clone cells.
METHODSThe bone marrow mononuclear cells (BMMNC) were extracted from 36 MDS patients and 14 normal controls. The mean fluorescence intensities (MFI) of phosphorylated STAT5(P-STAT5) in CD34(+)CD38(-)CD123(+) and CD34(+)CD38(-)CD123(-)cells, with or without the stimulation of 10 U/ml EPO, were examined by flow cytometry (FCM).
RESULTSWithout stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 113.71 ± 67.22/173.05 ± 102.78, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (58.84 ± 27.51/68.99 ± 50.42, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (63.06 ± 21.06, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; With the EPO stimulation, the P-STAT5 MFI in CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients was 144.04 ± 58.11/239.45 ± 152.05, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (68.41 ± 25, 10/64.21 ± 23.43, P < 0.01) and the normal controls CD34(+)CD38(-)CD123(-) cells (75.21 ± 27.02, P < 0.01), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal control CD34(+)CD38(-)CD123(-) cells; The P-STAT5 MFI in the CD34(+)CD38(-)CD123(+) cells of low/high risk MDS patients with or without EPO stimulation were 21.80/28.86, which was significantly higher than that of CD34(+)CD38(-)CD123(-) cells (7.42/5.50, P < 0.01, P < 0.05) and the normal controls CD34(+)CD38(-)CD123(-) cells (6.39, P < 0.05), there was no significant difference between the CD34(+)CD38(-)CD123(-) cells of MDS patients and the normal controls CD34(+)CD38(-)CD123(-) cells; There was no significant difference of P-STAT5 MFI with or without EPO stimulation and the increased P-STAT5 MFI between the CD34(+)CD38(-)CD123(+) cells of low and high risk MDS.
CONCLUSIONSTAT5 associated with cell proliferation was activated in CD34(+)CD38(-)CD123(+) bone marrow cells in MDS, which had more significant reactions to EPO than CD34(+)CD38(-)CD123(-) cells, indicating that CD34(+)CD38(-)CD123(+) bone marrow cells might be the real malignant MDS clone cells in MDS.
Adult ; Aged ; Aged, 80 and over ; Antigens, CD34 ; metabolism ; Bone Marrow Cells ; cytology ; metabolism ; Cell Proliferation ; Cells, Cultured ; Female ; Flow Cytometry ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; metabolism ; pathology ; Phosphorylation ; STAT5 Transcription Factor ; metabolism
6.Expression and clinical significance of Notch1 on the membrane of bone marrow CD38(+)CD138(+) plasma cells in the patients with multiple myeloma.
Yi-ran ZHAO ; Rong FU ; Jing GUAN ; Shan GAO ; Hui LIU ; Er-bao RUAN ; Wen QU ; Yong LIANG ; Guo-jin WANG ; Xiao-ming WANG ; Hong LIU ; Yu-hong WU ; Jia SONG ; Hua-quan WANG ; Li-min XING ; Jun WANG ; Li-juan LI ; Zong-hong SHAO
Chinese Journal of Hematology 2012;33(4):274-277
OBJECTIVETo investigate the expression of Notch1 on the membrane of bone marrow CD38(+)CD138(+) plasma cells in the patients with multiple myeloma (MM), and explore the importance of Notch signaling pathway in the formation and progression of MM.
METHODSThirty three MM patients and 15 healthy controls were enrolled in this study. The expression of Notch1 on the membrane of bone marrow CD38(+)CD138(+) and CD38(+)CD138(-) plasma cells were analyzed by flow cytometry. The clinical data of MM patients were also analyzed.
RESULTSThe ratio of Notch1 on the membrane of CD38(+)CD138(+) plasma cells of MM patients was (60.21 ± 25.06)% which was significantly higher than those of CD38(+)CD138(-) plasma cells of MM patients (39.84 ± 18.94)% (P = 0.000) and controls (38.34 ± 19.39)% (P = 0.004). There was no statistical difference between the two latter groups (P > 0.05). The expression of Notch1 on CD38(+)CD138(+)plasma cells from 24 newly diagnosed MM patients was correlated to the level of malignant plasma cells in there bone marrow (r = 0.914, P = 0.000), serum level of lactate dehydrogenase (LDH) (r = 0.754, P = 0.007), and β(2)-MG(r = 0.716, P = 0.013). The ratio of Notch1 on the membrane of CD38(+)CD138(+) plasma cells of MM patients who had renal dysfunction was correlated to their abnormal serum creatinine levels. The expression of Notch1 on CD38(+)CD138(+) plasma cells from 17 MM patients who received VD (bortezamib and dexamethasone) chemotherapy was correlated to the ratio of plasma cell reduction after the first VD chemotherapy (r = 0.842, P = 0.000).
CONCLUSIONThe expression of Notch1 on the membrane of CD38(+)CD138(+) plasma cells of MM patients was significantly higher than those of CD38(+)CD138(-) plasma cells of MM patients and controls. Notch1 overexpressed plasma cells were sensitive to the early VD therapy, and correlated to the progression and long term outcome of MM.
ADP-ribosyl Cyclase 1 ; immunology ; Adult ; Aged ; Aged, 80 and over ; Bone Marrow ; metabolism ; Case-Control Studies ; Cell Count ; Female ; Humans ; Male ; Middle Aged ; Multiple Myeloma ; immunology ; metabolism ; Plasma Cells ; immunology ; metabolism ; Prognosis ; Receptor, Notch1 ; metabolism ; Syndecan-1 ; immunology
7.Quantity and subtypes of dendritic cells in patients with immune related pancytopenia and their clinical significance.
Guang-Shuai TENG ; Rong FU ; Hui LIU ; Hong-Lei WANG ; Yi-Hao WANG ; Er-Bao RUAN ; Wen QÜ ; Yong LIANG ; Guo-Jin WANG ; Xiao-Ming WANG ; Hong LIU ; Yu-Hong WU ; Jia SONG ; Hua-Quan WANG ; Li-Min XING ; Jing GUAN ; Jun WANG ; Li-Juan LI ; Zong-Hong SHAO
Journal of Experimental Hematology 2012;20(3):722-726
This study was aimed to investigate the quantity and subtypes of dendritic cells (DC) in patients with immune related pancytopenia (IRP) and to explore the role of DC in pathogenesis of IRP. The quantity of plasmacytoid dendritic cells (pDC, Lin(-)HLA-DR(+) CD123(+) cells) and myeloid dendritic cells (mDC, Lin(-)HLA-DR(+) CD11c(+)cells) in peripheral blood of 65 patients with IRP (37 new diagnosed and 28 remitted) and 17 healthy controls were analyzed by flow cytometry. The results indicated that the ratio of pDC in peripheral blood mononuclear cells (PBMNC) was (0.91 ± 064)% in new diagnosed group, which was significantly higher than that in remission group (0.39 ± 0.11)% and control group (0.29 ± 0.13)% (P < 0.01), while this ratio of pDC in remission group was higher than that in control group (P < 0.05). The ratio of mDC in PBMNC was (0.21 ± 0.20)% in new diagnosed group and (0.34 ± 0.21)% in remission group respectively, there was no statistical difference as compared with control group (0.29 ± 0.09)% (P > 0.05). The ratio of pDC to mDC in new diagnosed group was 6.75 ± 7.11, which was significantly higher than that in remission group (1.55 ± 0.93) and control group (1.07 ± 0.43, P < 0.01), there was no statistical difference between the ratio of remission group and control group (P > 0.05). The ratio of pDC in PBMNC of IRP group negatively correlated to ratio of Th1/Th2 (r = -0.347, P < 0.05), and positively correlated to the ratio of auto-antibody on membrane of BMMNC (r = 0.606, P < 0.05) and to the quantity of CD5(+)B cells (r = 0.709, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.381, P < 0.01) and platelets (r = -0.343, P < 0.01). The ratio of mDC in PBMNC positively correlated to the ratio of Th1/Th2 (r = 0.595, P < 0.05) and the level of hemoglobin (r = 0.292, P < 0.05). The ratio of pDC/mDC negatively correlated to ratio of Th1/Th2 (r = -0.395, P < 0.05), it positively correlated to the level of antibody on membrane of BMMNC (r = 0.421, P < 0.05) and the quantity of CD5(+)B cells (r = 0.423, P < 0.05), while it negatively correlated to the levels of hemoglobin (r = -0.304, P < 0.05) and platelets (r = -0.287, P < 0.05). It is concluded that the quantity of pDC in peripheral blood of IRP patients increases, which may be related to the immunopathogenesis of IRP.
Adolescent
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Adult
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Blood Cell Count
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Case-Control Studies
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Child
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Child, Preschool
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Dendritic Cells
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cytology
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immunology
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Female
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Flow Cytometry
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Humans
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Male
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Middle Aged
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Pancytopenia
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blood
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immunology
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Young Adult
8.Application of bone marrow indirect Coombs test and its clinical significance in diagnosis of immuno-related pancytopenia.
Li-Juan BING ; Rong FU ; Hui LIU ; Hong-Lei WANG ; Er-Bao RUAN ; Wen QU ; Yong LIANG ; Guo-Jin WANG ; Xiao-Ming WNAG ; Hong LIU ; Yu-Hong WU ; Jia SONG ; Hua-Quan WANG ; Li-Min XING ; Li-Juan LI ; Zong-Hong SHAO
Journal of Experimental Hematology 2012;20(2):505-509
This study was purposed to applicate the bone marrow indirect Coombs test and investigate its clinical significancies in diagnosis of immuno-related pancytopenia (IRP). 30 patients with pancytopenia including 22 cases of IRP and 8 cases of idiopathic cytopenia of undetermined significance (ICUS), and 15 patients with iron-deficiency anemia as controls were enrolled in this study. After incubation of the bone marrow supernatant of IRP patients and bone marrow nucleated cell (BMNC) of controls was used as experiment group, while the incubation of BMNC and bone marrow supernatant of controls was used as control group. After incubation for 45 min, the positive rate of membrane antibodies in bone marrow hematopoietic cells (CD15(+), GlyCoA(+) and CD34(+)cells) was detected by flow cytometry, and correlation analysis of positive rate with clinical data of patients were analyzed. The results showed that among 30 patients with pancytopenia (16 positive and 14 negative for bone marrow direct Coombs test) 16 cases showed positive for bone marrow indirect Coombs test, with positive rate 53.33. In the experiment group, the median positive rate of CD15(+)IgM was 0.34, which was significantly higher than that in control group (0.20, P < 0.05); the median positive rates of CD34(+) IgG and IgM were 0.64 and 0.21 respectively, which were significantly higher than those in control group (0.00, P < 0.05) and (0.00, P < 0.05); the positive rates of GlyCoA(+)IgG and IgM were (0.83 ± 0.75) and (2.12 ± 1.98) respectively, which were significantly higher than those in control group [(0.47 ± 0.43), P < 0.05, (0.68 ± 0.64), P < 0.01]; the positive rates of CD15(+) IgG and IgM were positively correlated with the ratio of CD5(+)B cells. The positive rates of GlyCoA(+) IgG and IgM negatively correlated with the Hb level, percentage of reticulocytes, the ratio of bone marrow erythroid lineage and DC1/DC2 positively correlated with the ratio of CD5(+)B cells and indirect bilirubin level. It is concluded that antibodies (IgG or IgM) aiming at the bone marrow hematopoietic cells exist in the supernatant of some IRP and ICUS patients, and may act on the membrane protein of the normal BMNC. These antibodies correlate with the prognosis of IRP.
Adolescent
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Adult
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Aged
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Bone Marrow Cells
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immunology
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Child
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Coombs Test
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Female
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Humans
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Male
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Middle Aged
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Pancytopenia
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diagnosis
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immunology
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Young Adult
9.Quantity and function of T cell subsets in patients with paroxysmal nocturnal hemoglobinuria.
Tian ZHANG ; Yong LIANG ; Rong FU ; Li-Juan LI ; Jun WANG ; Hui LIU ; Hong-Lei WANG ; Er-Bao RUAN ; Wen QU ; Guo-Jin WANG ; Yu-Hong WU ; Hong LIU ; Hua-Quan WANG ; Xiao-Ming WANG ; Jia SONG ; Jing GUAN ; Li-Ming XING ; Zong-Hong SHAO
Journal of Experimental Hematology 2010;18(3):721-725
This study was purposed to investigate the immune state of T cells, the quantity and function of GPI(+) T cells and GPI(-) T cells in patients with paroxysmal nocturnal hemoglobinuria (PNH). 22 cases of PNH and 18 normal controls were enrolled in this study. Their T lymphocyte subsets, Th lymphocyte subsets were assayed by flow cytometry with the monoclonal antibodies concerned. The proportion of GPI(+) T cells or GPI(-) T cells in CD3(+) T cells, CD4(+) T cells, CD8(+) T cells and the expressions of CD69 on these T cells were also respectively assayed. The results showed that the proportion of CD4(+) T cells in CD3(+) T cells in PNH [(47.7670 +/- 13.91139)%] was lower than that in controls [(54.9592 +/- 7.11678)%] (p < 0.05). CD8(+) T cells in CD3(+) T cells of PNH cases [(52.2767 +/- 13.90395)%] were higher than that of controls [(45.2418 +/- 6.75306)%] (p < 0.05). The ratio of CD4(+) T cells to CD8(+) T cells was reverse in PNH. Those were more significantly in PNH-AA (0.77763 +/- 0.409153) (p < 0.05). The proportion of Th1 cells in PNH [(16.9136 +/- 6.78899)%], especially in PNH-AA [(22.8000 +/- 5.45244)%], was significantly higher than that in controls [(4.4600 +/- 1.81879)%] (p < 0.05). The proportion of Th2 cells in PNH [(4.7582 +/- 1.98441)%] had no difference from controls [(3.7960 +/- 1.13810)%]. The number of GPI(-) T cells in CD8(+) T cells and CD4(+) T cells were (14.6797 +/- 11.96718)% and (3.9241 +/- 2.46263)% respectively. The expression of CD69 on GPI(+) T cells or GPI(-) T cells in PNH [CD8(+) GPI(+) T cells (17.67881 +/- 8.562493)%, CD8(+) GPI(-) T cells (15.86575 +/- 7.279743)%, CD4(+) GPI(+) T cells (4.65431 +/- 1.984378)%, CD4(+) GPI(-) T cells (4.93181 +/- 1.730001)%]was significantly higher than that in normal controls [CD8(+) GPI(+) T cells (4.68038 +/- 1.216645)%, CD4(+) GPI(-) T cells (1.77339 +/- 0.645259)%] (p < 0.05), but the expression of CD69 on GPI(+) T cells was not different from that on GPI(-) T cells in PNH. It is concluded that high function of cytoimmunity in PNH may be responsible for bone marrow failure but not relates to the existence of PNH clone in T cell population.
Adolescent
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Adult
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Case-Control Studies
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Child
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Child, Preschool
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Female
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Flow Cytometry
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Hemoglobinuria, Paroxysmal
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immunology
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Humans
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Immunophenotyping
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Lymphocyte Count
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Male
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Middle Aged
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T-Lymphocyte Subsets
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immunology
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Young Adult
10.Perforin gene mutations in patients with acquired severe aplastic anemia.
Jian ZHANG ; Rong FU ; Jun WANG ; Li-Juan LI ; Jia SONG ; Wen QU ; Hua-Quan WANG ; Li-Min XING ; Hong LIU ; Yu-Hong WU ; Jin GUAN ; Guo-Jin WANG ; Xiao-Min WANG ; Yong LIANG ; Er-Bao RUAN ; Hui LIU ; Zong-Hong SHAO
Journal of Experimental Hematology 2011;19(2):431-434
This study was aimed to explore whether the perforin gene 1 (PRF1) mutation is the basis of genetic susceptibility to pathogenesis of acquired severe aplastic anemia (SAA). DNA exon2 and exon3 of PRF1 gene in peripheral blood mononuclear cells in 31 SAA patients and 15 normal controls were amplified by PCR; the sequencing was performed by using ABI pRISM 373OXL sequencer; the mutation loci were sought through checking sequences with GenBank-reported sequences; after the mutation sequences were found, those were cloned into M13 phage vector, then the corresponding sequences of gained 2 chromosomes were sequenced respectively to determine the distribution of different mutations on chromosomes. The results showed that (1) one homozygous mutation (822 C > T, synonymous mutation) and one heterozygous mutation (907 G > A, methionine 303 valine) were found in PRF1 coding region of 2 SAA patients. These mutations were not detected in normal controls. (2) 1 SNP (rs885822) in the coding region was detected in SAA patients and controls, and the heterozygosity rate between the 2 groups was different (p < 0.05). It is concluded that perforin gene mutation may be one risk factor in the aberrant proliferation and activation of cytotoxic T cells in pathogenesis of a part of patients with aplastic anemia.
Adolescent
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Adult
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Aged
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Anemia, Aplastic
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genetics
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Base Sequence
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Case-Control Studies
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Child
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Child, Preschool
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Female
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Genetic Predisposition to Disease
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Heterozygote
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Humans
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Male
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Middle Aged
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Mutation
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Perforin
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Pore Forming Cytotoxic Proteins
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genetics
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Young Adult