OBJECTIVETo explore the mechanism of 'erythroblast island (EI)' formation in the bone marrow of patients with immun-related hemocytopenia (IRP).

METHODSThe category of BM-auto antibody (au Ab) in 48 patients with IRP was detected with FCM. The BM-au Ab in the 'EI' of these cases were explored with immuonhistofluorescence (IF). Clinical and laboratory characteristics of these cases were also analyzed retrospectively.

RESULTSIgG could be detected in the 'EI' on the BM smear of 14 cases (29.17%), BM-au Ab mainly deposited at the edge/membranes between macrophage and erythroblasts rather than cyto plasm. Positive reaction were seen in all the cases with GlycoAIgG. The red blood cell count [(1.8 ± 0.5) × 10(12)/L] and hemoglobin level [(59.6 ± 16.2)g/L] were significantly lower than that in the IF(-) group [(2.5 ± 0.9) × 10(12)/L and (83.4 ± 25.0) g/L] (P < 0.05). The percentage of reticulocyte [(2.0 ± 0.8)%], serum level of IBIL [(9.4 ± 4.7) µmol/L], percentage of erythroblats in sternum BM (0.441 ± 0.139) and response rate to therapy (85.7%) in IF(+) group were significantly higher than that in IF(-)group [(1.3 ± 1.0)%, (6.6 ± 6.7)µmol/L, 0.298 ± 0.082, 61.3%, respectively] (P < 0.05).

CONCLUSIONMacrophage was connected with erythroblasts through autologous IgG in the 'EI's of some patients with IRP. 'EI' were the places where macrophages devoured and destroyed erythroblasts rather than erythroid development and differentiation. The pathogenetic mechanism of IRP might be associated with macrophages phagocytosing and destroying BM hematopoietic cells.

Blood Cell Count ; Bone Marrow ; Bone Marrow Cells ; immunology ; Coombs Test ; Erythroblasts ; Humans

Angiopoietin2( ANGPT2 ) plays an important role in tumor angiopoiesis. ANGPT2 antagonises ANGPT1 resulting in an effect on the stability of blood vessels, which promotes tumor growth, invasion, proliferation as well as relating to tumor vascular density. A lot of researches published papers about anti-ANGPT2 for the treatment of tumor, and have made some progresses. In this review, the role of ANGPT2 in the pathogenesis of acute myelogenous leukemia (AML), including its effects on proliferation of leukemia cells, bone marrow angiopoiesis, tumor invasion and metastasis are briefly summarised in order to provide the basis for targeted ANGPT2 in treatment of AML.
Angiopoietin-1 ; immunology ; Antibodies ; immunology ; Bone Marrow ; Humans ; Leukemia, Myeloid, Acute ; immunology ; Neoplasm Invasiveness ; Neoplasm Metastasis

OBJECTIVETo explore immunoregulatory mechanism of mesenchymal stem cells (MSCs) in H-2 haploidentical bone marrow cells transplantation mice.

METHODSBALB/c female mice irradiated with 8Gy 60Co gamma-rays were divided into two groups: MSCs group, infused cm-DiI labeled MSCs from female CB6F1 mice and monocytes from the bone marrow and spleen of male CB6F1; Control group, only infused monocytes from the bone marrow and spleen of male CB6F1. T-lymphocyte subpopulation of peripheral blood cells, T and B cells proliferation stimulated by ConA and LPS, mixed lymphocyte reaction between donor and recipient and third part, the sry-gene chimerism of bone marrow, spleen and thymus of the recipient, the distribution of MSCs in the recipient, the incidence rate of GVHD and survival were observed.

RESULTSThe CD3 at +90 d the percent of CD3+ CD4+ cells, and CD4/CD8 at +30 d in the MSCs group were higher than that in control post-transplantation, respectively (P < 0.05). The proliferation activity of B cells recovered more rapidly and that of T cells recovered comparably in MSCs group as compared with that in control group. The result of MLR between donor and recipient was lower in MSCs group than that in the control; and that between recipient and the third part had no difference. The sry-gene chimerism of bone marrow and spleen of the recipient was higher in MSCs group than in control at +30 d. The MSCs mainly distributed in intestine, thymus, bone marrow, liver, heart of the recipient after transplantation. The incidence of acute GVHD was higher and the survival rate was lower in MSCs group than that in control group (P < 0.05). Chronic GVHD occurred in the control group at +90 d, while in the MSCs group at +120 d.

CONCLUSIONSMSCs might improve stem cell engraftment, promote lymphocyte and humoral immunity recovery, decrease incidence of GVHD and increase survival by inducing specific immunologic tolerance and repairing organs injuries.

Animals ; Bone Marrow Cells ; immunology ; Bone Marrow Transplantation ; immunology ; Female ; Graft vs Host Disease ; immunology ; Male ; Mesenchymal Stromal Cells ; immunology ; Mice ; Mice, Inbred BALB C

The purpose of this study was to detect the distribution of Treg and Th17 cells in bone marrow and to investigate the relationship of Treg/Th17 imbalance with the pathogenesis and progression of multiple myeloma (MM). The Bone marrow was collected from 37 MM patients and 12 healthy volunteers, the ratio of Treg and Th17 cells was detected by flow cytometry. The expression of Treg and Th17 cells simultaneously was examined in peripheral blood of 19 MM patients with same method. The results indicated that the frequency of Treg cells was higher in MM patients than that in control group (P < 0.05), there was a trend of increasing of Treg cell number in the ISS stage from I+II to III (P < 0.05). Furthermore, in the patients with MM, the Treg cell number in bone marrow was higher than that in peripheral blood (P < 0.05). Th17 cell rate was not statistically different between MM patients and control group (P > 0.05), and at different ISS stage (P > 0.05). Th17 cell number between bone marrow and peripheral blood was not significantly different (P > 0.05).The ratio of Treg/Th17 in patients with MM was higher than that in control group (P < 0.05), and increased gradually from ISS stage I+II to stage III (P < 0.05). It is concluded that the Treg/Th17 immune imbalance is presenced in bone marrow of patients with MM, this imbalance may promote the progression of MM.
Bone Marrow ; immunology ; Cell Count ; Disease Progression ; Flow Cytometry ; Humans ; Multiple Myeloma ; immunology ; T-Lymphocytes, Regulatory ; immunology ; Th17 Cells ; immunology

OBJECTIVETo explore the pathogenetic mechanism of cytopenia in the patients with systemic lupus erythematosus (SLE).

METHODSTwenty-one patients with SLE and 10 healthy controls were enrolled in this study. Bone marrow mononuclear cells (BMMNC) Coombs test was used to determine the autoantibodies and in vitro CFU-E, BFU-E and CFU-GM yields were used to determine the proliferation function of hematopoietic stem/progenitor cell (HSC).

RESULTSPositive results of BMMNC-Coombs test were observed in 12 patients with SLE (57.1%), among them, 10 with (58.2%) and 2 without blood cytopenia. The yields of CFU-GM (49 +/- 40), and CFU-E (49 +/- 35) in SLE patients without cytopenia were higher than that in normal controls, but lower than that in cytopenia SLE patients. The BFU-E yield in SLE patients (3 +/- 4) was significantly lower than that in normal controls (36 +/- 12).

CONCLUSIONThere were auto-antibodies on BMMNC in patients with SLE. The blood cytopenia in SLE patients maybe resulted from the destructions of bone marrow hematopoietic cells by the autoantibodies. The capacities of bone marrow HSC in SLE patients for proliferation compensation of hematopoietic cell destruction were not damaged.

Adolescent ; Adult ; Autoantibodies ; blood ; Bone Marrow ; immunology ; pathology ; Bone Marrow Cells ; immunology ; pathology ; Case-Control Studies ; Female ; Humans ; Lupus Erythematosus, Systemic ; immunology ; pathology ; Male ; Middle Aged ; Young Adult

OBJECTIVETo explore the hematopoietic stem cell distribution and lymphocyte proliferation and differentiation in recipient mice after allogeneic bone marrow transplantation (allo-BMT).

METHODSBALB/c (H-2(d)) mice were total body irradiated 5.5 Gy x 2 by (137)Cs and then transplanted with bone marrow cells from GFP transgenic C57BL/6J (H-2(b)) mice. The femur, spleen, Peyer patches, thymus, liver and peripheral blood of the host were collected on days 3, 7, 21, 35 and 70 post transplantation, and their sections were observed by fluorescent microscopy. The green fluorescent cells were counted with FACS. The phycoerythrin (PE) labeled antibodies to CD4, CD8 and B220 were used for sorting T and B lymphocytes.

RESULTS(1) On day 3 and day 7 after allo-BMT, there were (1.06 +/- 0.02)% and (76.60 +/- 1.80)% of donor's green bone marrow cells in host's spleen respectively, whereas only (0.37 +/- 0.06)% and (39.70 +/- 5.38)% in the bone marrow, respectively. (2) In bone marrow and other organs of 21 day-old chimerism mice, over 60% cells were of donor origin. (3) There were (0.36 +/- 0.04)% donor's bone marrow cells lodging at host's Peyer patches, similar to that in bone marrow.

CONCLUSION(1) The engrafted allogeneic hematopoietic stem cell can move into spleen, bone marrow, Peyer patches and thymus. The spleen is the main lodging place of the engrafted cells early after all-BMT. (2) The majority of cells in chimerism mice immunologic organs were of donor origin. (3) Peyer patches is another lodging place early after allo-BMT.

Animals ; Bone Marrow Cells ; immunology ; Bone Marrow Transplantation ; immunology ; Lymphocytes ; immunology ; Mice ; Mice, Inbred BALB C ; Mice, Inbred C57BL ; Tissue Donors

Bone Marrow ; immunology ; pathology ; Drugs, Chinese Herbal ; therapeutic use ; Hematopoietic System ; immunology ; pathology ; Humans ; Immunosuppression ; Phytotherapy

This study was purpose to investigate the biological characteristics of B lymphoblastic leukemia (B-ALL) between CD34 positive CD38 positive (CD34(+)CD38(+)) and CD34(+)CD38(low/-) subgroups and their clinical significance. Immunophenotyping of B cells in bone marrow of 54 patients with newly diagnosed CD34(+)B-ALL were analyzed by 4 color multiparametric flow cytometry (FCM). According to the different expression of CD38, the newly diagnosed patients with B-ALL were divided into two groups: CD34(+)CD38(+) subgroup and CD34(+)CD38(low/-) subgroup. BCR-ABL, TEL-AML1 fusion genes and WT1 gene were detected by real time RT-PCR simultaneously. After chemotherapy, minimal residual disease (MRD) was monitored by one tube of 7 color FCM. The average follow-up time was 12 months (range 1 - 28), the average follow-up interval was 2 months (range 1 - 5). The results showed that there was no significant differences such as WBC, Plt count and Hb level between the two groups at diagnosis, the positive rate of BCR-ABL, TEL-AML1 and WT1 gene was also no significantly different. After clinical complete remission (CR), MRD positive (MDR(+)) case rates were 28.57% (10/35) in CD34(+)CD38(+) subgroup and 68.42% (13/19) in CD34(+)CD38(low/-) subgroup (P < 0.01). The relapse rate between the two groups was 5.71% (2/35) in CD34(+)CD38(+) subgroup (relapse time at 94 and 245 d respectively) and 36.84% (7/19) in CD34(+)CD38(low/-) group [median relapse time was 263 d (range 46 - 468), P < 0.01]. The age distribution was analyzed in these two subgroups (> 16 or ≤ 16 years old), there was 8 (8/35) adult patients (> 16 years old) in CD34(+)CD38(+)group and 10 (10/19) adult patients in CD34(+)CD38(low/-) group (P < 0.05). It is concluded that CD34(+)CD38(low/-) phenotype is more often presented in adult patients and the CD34(+)CD38(low/-) patients with B-ALL are more likely to have MRD(+)and relapse after treatment.
ADP-ribosyl Cyclase 1 ; immunology ; Adolescent ; Adult ; Antigens, CD34 ; immunology ; Bone Marrow ; immunology ; Bone Marrow Cells ; immunology ; Child ; Child, Preschool ; Female ; Flow Cytometry ; Humans ; Immunophenotyping ; Infant ; Leukemia, B-Cell ; immunology ; Male ; Middle Aged ; Neoplasm, Residual ; immunology ; Young Adult

OBJECTIVETo study the quantity and ratio of Th1, Th2 cells in the bone marrow of myelodysplastic syndromes (MDS) patients, and to evaluate the correlation between the ratio of the blast cells and the number of the Th1 cells in the bone marrow of MDS patients.

METHODSBy FACS, the quantity and ratio of IFN-gamma producing CD4(+) T cell (Th1) and IL-4 producing CD4(+) T cell (Th2) cells in the bone marrow were detected in 21 MDS patients, 18 normal controls and 13 severe aplastic anemia (SAA) patients respectively. The karyotypes of 18 MDS patients and 15 normal controls were assayed. The correlation between the ratio of the blast cells in the bone marrow and the number of the Th1 cells in the MDS patients were analyzed.

RESULTSThe percentages of Th1 cells, Th2 cells and ratio of Th1/Th2 in the bone marrow of normal controls were (0.48 +/- 0.10)%, (0.24 +/- 0.19)% and 2.31 +/- 0.76 respectively, while those of the MDS patients were (0.36 +/- 0.11)%, (0.76 +/- 0.35)% and 0.51 +/- 0.13. The percentage of Th1 cells of patients with MDS was reduced and the Th1/Th2 ratio was significantly lower than that of normal controls (P < 0.01). Those of the patients with SAA were (4.75 +/- 0.49)%, (0.40 +/- 0.28)% and 26.5 +/- 8.79 respectively, their Th1 cells and Th1/Th2 ratio were markedly higher than those of normal controls (P < 0.01). In all of the 15 normal controls the karyotypes were normal, but that of MDS patients was (50.00 +/- 0.10)%. The lower ratio of the Th1 cells in the bone marrow of the patients with MDS and the AML which progressed from MDS was negatively correlated with the higher percentage of the blast cells (r = -0.563, P < 0.01).

CONCLUSIONS(1) The immune function of T lymphocytes in MDS is abnormal: the balance between Th1 and Th2 cells is broken. (2) With descending of the number of Th1 cells in the bone marrow of the MDS patients, the disease is progressing to leukemia.

Adult ; Aged ; Bone Marrow ; immunology ; Female ; Humans ; Karyotyping ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; immunology ; T-Lymphocytes, Helper-Inducer ; immunology

This study was purposed to explore the immunoregulatory effects of human bone marrow mesenchymal stem cells (MSCs) on active T lymphocytes in vitro and the new strategy to prevent graft-versus-host disease (GVHD) in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Mononuclear cells from human peripheral blood cells were isolated and cultured in the presence of phytohemagglutinin (PHA) (final concentration was 10 microg/ml) for different times. The ability of T lymphocyte proliferation and activation was measured by (3)H-Thyramine incorporation. The expressions of CD3(+)CD4(+), CD3(+)CD8(+), CD4(+)CD25(+) and CD4(+)CD152(+) on T cells were detected by FCM after coculture for 72 hours. Experiment was divided into 4 groups: A group as control (no added MSCs), B group (actived T cells + 2 x 10(4) MSCs), C group (actived T cells + 4 x 10(4) MSCs), D group (actived T cells + 8 x 10(4) MSCs). The results showed that the ability of T lymphocyte proliferation in the same PHA concentration increased with prolonging of time. ability of T lymphocyte proliferation was strongest when culturing for 48 hours (p < 0.01); the expressions of CD44, CD105, CD29 and FIK1 of MSCs were positive, expressions of CD33, CD34, CD45 and HLA-DR were negative. MSCs inhibited T lymphocyte proliferation and the inhibitory effect depended on the amount of MSCs. CD3(+)CD8(+), CD4(+)CD25(+) and CD4(+)CD152(+) T cells cocultured with MSCs increased obviously and CD3(+)CD4(+) expression significantly decreased, as compared with control group (p < 0.01). It is concluded that the MSCs inhibit T lymphocyte proliferation induced by mitogen (PHA), and perform their immunosuppressive function by up-regulation of CD3(+)CD8(+), CD4(+)CD25(+) and CD4(+)CD152(+) expressions and down-regulation of CD3(+)CD4(+) expression.
Bone Marrow Cells ; cytology ; Cell Separation ; Cells, Cultured ; Humans ; Lymphocyte Activation ; immunology ; Mesenchymal Stromal Cells ; cytology ; immunology ; T-Lymphocytes ; immunology