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Primary mediastinal large B-cell lymphoma (PMBCL) is an aggressive large B-cell lymphoma originating in the mediastinum. Because of its distinct clinical and histological features, PMBCL has been reclassified as a separate entity by the World Health Organization classification of lymphoid neoplasms. The diagnosis of PMBCL mainly depends on the pathological features, imaging examination and clinical features. Currently, there are many therapeutic schemes for PMBCL, the most commonly used schemes are R-CHOP and R-EPOCH regimens. Radiotherapy is beneficial in some patients, but it can also lead to long-term toxicity. Research and development of new drugs are ongoing, including chimeric antigen receptor T-cell therapy, anti-programmed death receptor 1 drugs, etc. PET-CT is mainly used to assess the curative effect after treatment and to guide the next treatment strategy.
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Aplastic anemia (AA) is a bone marrow failure syndrome characterized by pancytopenia. Decreased numbers of hematopoietic stem cells and impaired bone marrow microenvironment caused by abnormal immune function describe the major pathogenesis of AA. Hematopoietic stem cell transplantation and immunesuppressive therapy are the first-line treatments for AA. Porcine anti-lymphocyte globulin (p-ALG) is a new product developed in China. Several studies have shown that p-ALG exhibited good therapeutic effects in AA.
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Anemia Aplásica , Globulinas , Trasplante de Células Madre Hematopoyéticas , Anemia Aplásica/tratamiento farmacológico , Animales , Células Madre Hematopoyéticas , Inmunosupresores , Linfocitos , PorcinosRESUMEN
Objective@#To investigate the levels of NK cells and their relevant cytokines (IL-10, TGF-β and IFN-γ) in patients with primary immune thrombocytopenia (ITP) .@*Methods@#All samples were obtained from 42 patients (22 newly diagnosed and 20 in remission) and 20 healthy volunteers. The levels of IL-10 and IFN-γ in blood serum were detected by enzyme-linked immunosorbent assay (ELISA) . The percentage of CD3- CD56+ NK cell, CD3- CD56bright CD16- NK cell, CD3- CD56dim CD16+ NK cell in peripheral blood lymphocyte were detected by flow cytometry. The NK cells were isolated by immunomagnetic microbeads. The mRNA expression levels of IL-10, TGF-β, and IFN-γ in NK cells were detected by real-time fluorescent quantitative PCR. Correlation between the above measured results was analyzed.@*Results@#① The blood serum level of IFN-γ in newly diagnosed ITP patients [ (653.0±221.6) ng/L] was higher than that in remission ITP patients [ (484.4±219.5) ng/L] and healthy control [ (390.9±253.5) ng/L] (P=0.022, P=0.001) . The blood serum level of IL-10 in newly diagnosed ITP patients was lower than that in healthy control [ (52.09±26.66) ng/L vs (79.44±38.43) ng/L, P=0.007]. ②The percentage of NK cell in newly diagnosed and remission ITP patients [ (9.53±3.93) %, (9.03±3.78) %] were significantly lower than that in healthy control [ (13.72±7.42) %] (P=0.013, P=0.007) . The ratio of CD3- CD56bright CD16- NK cell/total NK cells in newly diagnosed ITP patients was higher than that in healthy control [ (6.85±4.43) % vs (4.05±2.81) %, P=0.032]. The ratio of CD3-CD56dim CD16- NK cell/total NK cells in newly diagnosed ITP patients was lower than that in healthy control [ (93.14±4.43) % vs (95.94±2.81) %, P=0.032]. ③ There was no significant difference in the mRNA expression level of IFN-γ in NK cells of ITP patients and healthy control (all P>0.05) . The mRNA expression levels of IL-10 and TGF-β in NK cells in newly diagnosed ITP patients were significantly higher than that in healthy control (1.82±1.32 vs 1.02±1.03, P=0.023; 2.80±2.31 vs 1.46±1.37, P=0.028) . The ratio of CD3-CD56bright CD16- NK cell/total NK cells was positively correlated with the mRNA expression levels of IL-10, TGF-β in NK cells (r=0.424, P=0.001; r=0.432, P<0.001) .@*Conclusion@#NK cells may compensate for the deficiency of the number by enhancing the secretion of negative regulation cytokines, acting as "protective" roles in the disease.
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Objective@#To investigate the change of autophagy level of bone marrow nucleated red blood cell (RBC) in patients with myelodysplastic syndromes (MDS) .@*Methods@#Fifty-four MDS patients and thirty-three controls were enrolled in this study. The mitophagy were observed by transmission electron microscopy (TEM) . The level of autophagy-associated protein LC3B in GlycoA+ nucleated RBC was measured by flow cytometry. The expressions of ULK1 and mTOR mRNA in GlycoA+ nucleated RBC were measured by real-time PCR. The expression of the mitochondrial outer membrane protein TOM20 in GlycoA+ nucleated RBC was detected by Western blot.@*Results@#Autophagosomes or autolysosomes were scarcely observed by TEM in MDS patients. The expression of LC3B in GlycoA+ nucleated RBC in high-risk MDS patients (0.22±0.12) was significantly lower than that in normal controls (0.43±0.22, P<0.001) , and lower than that in low-risk MDS patients (0.40±0.16, P=0.001) . The expression of AMPK [0.26 (0.60) ] in GlycoA+ nucleated RBC in high-risk MDS patients was significantly lower than that in controls [1.00 (2.07) , P<0.017) . The expression of ULK1 mRNA in GlycoA+ nucleated RBC in high-risk MDS patients [0.27 (3.31) ] was significantly lower than that in controls [1.07 (4.41) , P<0.017]. The level of mTOR mRNA in GlycoA+ nucleated RBC in high-risk MDS patients [1.82 (3.74) ] was significantly higher than that in controls [1.26 (1.38) , P<0.017]. The level of LC3B in GlycoA+ nucleated RBC was negatively correlated with the HGB (r=0.529, P=0.009) in high-risk MDS patients. The expression of mitochondrial outer membrane protein TOM20 in high-risk MDS patients was 9.42±4.42.@*Conclusion@#Autophagy is impaired in nucleated RBC of MDS patients.
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Objective@#To explore characteristic and function of peripheral blood mononuclear cells (PBMNC) -induced macrophages in patients with myelodysplastic syndrome (MDS) to couple with its progression.@*Methods@#A total of 24 MDS patients (11 low-risk patients and 13 high-risk group patients) referred to Department of Hematology of Tianjin Medical University General Hospital and normal controls were enrolled from September 2014 to December 2015. PBMNC was stimulated with GM-CSF to transform to macrophages. The morphology of macrophages was observed by microscope. The quantity of macrophages, CD206 and SIRPα on surface of macrophages were detected by flow cytometry. The phagocytic function of macrophages was analyzed by fluorescence microscopy and flow cytometry.@*Results@#The morphology of macrophages from MDS patients was abnormal. The percentage of transformed macrophages was (5.17±3.47) % in patients with MDS, which was lower than that in controls significantly[ (66.18±13.43) %, t=3.529, P=0.001]. The expression of CD206 on macrophages from MDS patients was significantly lower than that of controls[ (9.73±2.59) % vs (51.15±10.82) %, t=4.551, P<0.001]. The SIRPα level of macrophages from MDS patients was significantly lower than that of controls [ (0.51±0.09) % vs (0.77±0.06) %, t=2.102, P=0.043]. The phagocytic index and the percentage of phagocytic of macrophages from MDS patients were significantly lower than those of macrophages from normal controls[0.45±0.08 vs 0.92±0.07, t=-6.253, P=0.008; (23.69±3.22) % vs (42.75±2.13) %, t=-6.982, P=0.006 respectively]by flow cytometry. The phagocytic index of MDS patients was significantly lower than that of controls (0.24±0.04 vs 0.48±0.96, t=3.464, P=0.001) by fluorescence microscopy.@*Conclusion@#The quantity, recognization receptors and phagocytosis of PBMNC-induced macrophages decreased in MDS patients.
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BACKGROUND: Prolymphocytic leukemia (PLL) is a generalized malignancy of the lymphoid tissue, usually of B cell type. Auer rod-like inclusions in prolymphocytic leukemia cells are an extremely rare event; the inclusions are very similar to the Auer rods morphologically. METHODS: We describe a case of B-cell PLL presenting with Auer rod-like inclusions. The diagnosis was eventually proven by the morphology, cytochemical staining, immunophenotypes, and electron microscopy. RESULTS: Auer rod-like inclusions are pathological changes of mitochondria with increasing density of matrix and disappearing internal instructure seen through a scanning electronic microscope. CONCLUSIONS: Auer rod-like inclusions can present in pathologically changed prolymphocytic leukemia cells.
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Linfocitos B/ultraestructura , Leucemia Prolinfocítica Tipo Células B/patología , Mitocondrias/ultraestructura , Linfocitos B/inmunología , Biomarcadores de Tumor/análisis , Examen de la Médula Ósea , Humanos , Inmunofenotipificación , Leucemia Prolinfocítica Tipo Células B/inmunología , Masculino , Microscopía Electrónica de Rastreo , Persona de Mediana Edad , Mitocondrias/inmunología , Valor Predictivo de las PruebasRESUMEN
<p><b>OBJECTIVE</b>To investigate the change of autophagy level of bone marrow mononuclear cells(BMMNCs)in patients with myelodysplastic syndromes(MDS).</p><p><b>METHODS</b>Thirty- eight patients with MDS and 26 megaloblastic anemia patients were enrolled in this study. The autophagic vacuoles were observed by transmission electron microscopy (TEM) and the quantity of autophagic vacuoles was detected by monodansylcadaverine (MDC) staining. The LC3 protein positive cells were counted by immunofluorescence assays. The expression of Beclin 1, LC3A, mTOR mRNA were measured by real time PCR. The expression of Beclin 1 proteins were detected by Western blotting.</p><p><b>RESULTS</b>The autophgic vacuoles of double membrane that surrounds lysosomes appeared in MDS patients. The percentage of MDC positive cells was significantly higher in MDS patients[(9.75±2.63)%]than that of controls[(2.90± 0.89)%, P<0.05). The percentage of LC3 protein cells was also increased in MDS patients(6.13±1.03)% vs(1.5±0.58)%, P<0.05). The expression of Beclin 1 and LC3A mRNA in low-risk and intermediate-1 MDS were higher compared with controls (3.61 ± 3.02 vs 1.55 ± 1.03 and 6.56 ± 3.97 vs 1.21 ± 0.95 respectively, both P<0.05). The expression of mTOR mRNA was down- regulated in low- risk and intermediate-1 MDS compared with controls(0.39±0.37 vs 1.50±1.03, P<0.05). There were no significant difference in expression of Beclin 1, LC3 and mTOR mRNA among intermediate-2 and high-risk MDS and controls. Beclin 1 protein expression was higher in low- risk and intermediate- 1 MDS patients(1.257 ± 0.197)than that of controls(0.528±0.086)and inermediate-2 and high-risk MDS patients(0.622±0.118).</p><p><b>CONCLUSION</b>The autophagy levels were increased in low- risk and intermediate- 1 MDS, while not enhanced in intermediate-2 MDS. Autophagy might be considered as a cell protective mechanism in MDS. The relatively defective autophagy in intermediate- 2 and high- risk MDS might contribute to disease's progression.</p>
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Humanos , Proteínas Reguladoras de la Apoptosis , Metabolismo , Autofagia , Beclina-1 , Células de la Médula Ósea , Biología Celular , Proteínas de la Membrana , Metabolismo , Microscopía Electrónica de Transmisión , Proteínas Asociadas a Microtúbulos , Metabolismo , Síndromes Mielodisplásicos , Patología , Serina-Treonina Quinasas TOR , Metabolismo , VacuolasRESUMEN
<p><b>OBJECTIVE</b>To study the expression of CD70 and the methylation level of CD70 promoter in immuno-related pancytopenia(IRP)patients, and explore the role of CD70 in the pathogenesis of IRP.</p><p><b>METHODS</b>Thirty- five IRP patients and fifteen healthy donors were enrolled in this study. Peripheral blood mononuclear cells were isolated from venous blood by density gradient centrifugation, and CD4(+) T cells were isolated by immunomagnetic beads. The mRNA level and the percentage of CD70 of CD4(+) T cells were measured by real- time quantitative polymerase chain reaction(RT- PCR)and flow cytometry respectively. Methylation- Specific PCR(MSP)was performed to determine the methylation level of CD70 promoter.</p><p><b>RESULTS</b>The percentage of CD70 expression on CD4(+) T cells of untreated IRP patients[(7.46±1.51)%]was significantly higher than that of recovered ones[(5.95±1.34)%]and normal controls[(1.83 ± 0.60)%], and that of recovered IRP patients was significantly higher than of normal controls(P<0.05). The relative expressions of CD70 mRNA in CD4(+) T cells were 2.314(0.200-6.084), 1.021(0.135-3.434), 0.353(0.008-2.258)in three groups respectively. The differences among untreated IRP patients, recovered IRP patients and normal controls were significant(P<0.05). The CD70 promoter methylation level in CD4(+) T cells of all IRP patients was significantly lower than that of normal controls (P<0.05). The expression of CD70 positively correlated to the ratio of CD5(+) B cells(r=0.533, P<0.01).</p><p><b>CONCLUSION</b>The overexpression of CD70 may lead to immunologic disarrangement of patients, which may play important role in the pathogenesis of IRP.</p>
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Humanos , Linfocitos B , Ligando CD27 , Linfocitos T CD4-Positivos , Citometría de Flujo , Pancitopenia , Regiones Promotoras Genéticas , ARN Mensajero , Reacción en Cadena en Tiempo Real de la PolimerasaRESUMEN
<p><b>OBJECTIVE</b>To explore the expression levels of terminal complement complex (C5b-9) and CD62p on platelets and the soluble C5b-9 (sC5b-9) level in serum in patients with PNH or PNH-aplastic anemia (AA).</p><p><b>METHODS</b>Serum levels of sC5b-9, complement C3 and C4 were detected by using ELISA in 25 patients with PNH/PNH-AA. The quantities of C5b-9 and CD62p on the membrane of platelets were detected by flow cytometry.</p><p><b>RESULTS</b>①In PNH/PNH-AA group, the serum sC5b-9 level [390.27(265.73-676.87) μg/L] was lower than that in control group [540.39(344.20-1 576.78) μg/L] (P<0.01). ②The platelet PNH clone (CD59⁻CD61⁺/CD61⁺) size [50.58(23.29-81.60)%] was bigger in the PNH/PNH-AA group than that [23.57(15.58-29.02)%] in control group (P<0.01). The percentages of C5b-9 deposition (C5b-9⁺CD61⁺/CD61⁺) were higher on the PNH clone platelets (CD59⁻CD61⁺) in the PNH/PNH-AA group [(17.53 ± 6.27)%] than those on the normal platelets (CD59⁺CD61⁺) in PNH patients 11.33±5.03)%] and control [(10.88±3.58)%] group (P<0.01). ③ The expression of CD62p (CD62p⁺CD61⁺/CD61⁺) on PNH clone platelets in PNH patients [(61.98 ± 11.71)%] was higher than that on the normal platelets in PNH patients [(43.76±11.30)%] and control group [(38.23±18.07)%] (P<0.01). In addition, the expression of CD62p on normal platelets was higher in PNH patients than control (P<0.05). ④The deposition of C5b-9 positively correlated with the expression of CD62p on the platelets (r=0.559, P=0.002).</p><p><b>CONCLUSION</b>Deficiency of CD59 antigen on platelets in PNH patients may lead to the deposition of C5b-9 on its membrane and its dysfunction, which may contribute to thrombosis events in PNH.</p>
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Humanos , Anemia Aplásica , Plaquetas , Células Clonales , Complejo de Ataque a Membrana del Sistema Complemento , Citometría de Flujo , Hemoglobinuria Paroxística , Selectina-P , TrombosisRESUMEN
Abnormal epigenetics play important roles in the pathogenesis of myelodysplastic syndromes (MDS). DNA hypermeth-ylation is the most common epigenetic abnormality in MDS. Demethylating DNA hypermethylation may improve the quality of life of MDS patients and prolong their overall survival. Azacitidine and decitabine are the demethylating drugs approved for MDS treatment. These drugs showed clinical effects on all subgroups of MDS patients.
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<p><b>OBJECTIVE</b>To detect memory B lymphocyte (Bm) in peripheral blood (PB) of immune-related pancytopenia (IRP).</p><p><b>METHODS</b>86 patients with IRP and 11 health volunteers were enrolled in this study. Bm (CD5⁺ CD19⁺ CD27⁺) and bone marrow mononucleated cell antibodies (BMMNC-Ab) were determined via fluorescence-activated cell sorting, and clinical outcomes of these patients were analyzed.</p><p><b>RESULTS</b>(1)43 initial patients achieved obvious remission in all 52 initial cases after conventional immunosuppression therapy. 16 relapsed patients with IRP received Rituximab (RTX) and 14 cases achieved obvious remission, among which 7 cases were refractory to conventional immunosuppression therapy, 5 cases exhibited obvious remission, and 2 cases did not respond. Other 18 relapsed cases received conventional immunosuppression therapy and 13 cases achieved obvious remission. (1)The level of Bm in PB in 52 initial patients with IRP was(1.81 ± 0.97)%, and no significant difference was observed between the initial patients and health volunteers (1.75 ± 0.55)% (P>0.05). The level of Bm in PB in 34 relapsed patients with IRP was obviously higher than that in the initial IRP patients and health volunteers (P<0.05). Significant difference was observed in the level of Bm in PB in 16 relapsed IRP patients between pre-therapy and post-therapy with RTX (P<0.05). No statistical difference was found between the remission and no-response groups in relapsed patients treated with RTX. RTX regimen produced more effective outcome than conventional immunosuppression therapy, which better eliminated Bm than the latter (P<0.05). Initial patients with IRP who relapsed within a two-year follow-up period had a lower level of Bm in PB compared with un-relapsed patients (P<0.05). Majority of BMMNC- Ab antibodies in relapsed patients were IgG (82.4%) and IgM (69.2%) autoantibodies in patients with initial IRP.</p><p><b>CONCLUSION</b>The level of Bm in PB was associated with relapsed patients with IRP. Bm did not respond to conventional immunosuppression therapy,but responded to RTX.</p>
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Adolescente , Adulto , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Anticuerpos Monoclonales de Origen Murino , Usos Terapéuticos , Subgrupos de Linfocitos B , Alergia e Inmunología , Memoria Inmunológica , Terapia de Inmunosupresión , Pancitopenia , Alergia e Inmunología , Terapéutica , Recurrencia , Rituximab , Resultado del TratamientoRESUMEN
<p><b>OBJECTIVE</b>To culture osteoblast in vitro and evaluate CCL3 receptor CCR1 expression in patients with multiple myeloma (MM).</p><p><b>METHODS</b>Bone marrow osteoblasts from MM patients were cultured in vitro with dexamethasone, β-sodium glycerophosphate and vitamin C, which were identified by alkaline phosphatase staining, Von Kossa's staining. The CCL3 receptor expression was evaluated by flow cytometry. The morphology and quantity of osteoblast were observed after exposure to CCL3.</p><p><b>RESULTS</b>Bone marrow osteoblasts from MM patients could be cultured in vitro and be identified by positive staining of alkaline phosphatase and Von Kossa's. MM-derived osteoblasts expressed higher levels of CCR1 (74.48 ± 7.31)%, compared with normal controls (48.35 ± 8.81)%. Calcium deposition of osteoblasts after exposure to CCL3 was less than that of controls.</p><p><b>CONCLUSION</b>Bone marrow osteoblasts could be cultured in vitro from MM Patients. CCL3 may contribute to the development of myeloma bone disease.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Células Cultivadas , Quimiocina CCL3 , Farmacología , Mieloma Múltiple , Patología , Osteoblastos , Biología Celular , Metabolismo , Receptores CCR1 , MetabolismoRESUMEN
<p><b>OBJECTIVE</b>To investigate the changes of relative telomere length (RTL) of peripheral blood (PB) CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺T lymphocytes, CD19⁺B lymphocytes and bone marrow (BM) CD34⁺ cells and its association with disease severity in untreated patients with immuno-related pancytopenia (IRP).</p><p><b>METHODS</b>The PB CD3⁺ , CD3⁺ CD4⁺ , CD3⁺ CD8⁺ T lymphocytes, CD19⁺ B lymphocytes, and BM CD34⁺ cells were purified by magnetic activated cell sorting (MACS), and RTL were measured with flow-fluorescence in situ hybridization (FLOW-FISH).</p><p><b>RESULTS</b>The RTL of CD3⁺, CD3⁺CD4⁺ , and CD3⁺CD8⁺T lymphocytes in untreated IRP patients were (27.754 ± 16.323)%, (7.526 ± 3.745)% and (25.854 ± 14.789)%, respectivly, which were significantly shorter than those in healthy-controls (54.555 ± 19.782)%, (12.096 ± 2.805)%, and (38.367 ± 4.626)% (P<0.05). The RTL of CD19⁺ lymphocytes in untreated IRP patients was (22.136 ± 16.142)%, which was significantly shorter than that in healthy controls (42.846 ± 16.353)% (P<0.01). There was no significant difference of BM CD34⁺ cells RTL between the untreated IRP patients (22.528 ± 21.601)% and the healthy controls (23.936 ± 19.822)% (P>0.05). There were significantly positive correlations between the RTL of B lymphocytes and the count of white blood cell (r=0.706, P=0.015). There were negative correlations between RTL of B lymphocytes and the clinical symptoms (r=-0.613, P=0.045) and positive correlations with therapeutic effect (r=0.775, P=0.005).</p><p><b>CONCLUSION</b>The shorter RTL of CD3⁺, CD3⁺CD4⁺, CD3⁺CD8⁺, CD19⁺ lymphocytes, and the normal RTL of BM CD34⁺ cells in untreated IRP patients were identified, which might imply that IRP is a type of acquired autoimmune diseases.</p>
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Adolescente , Adulto , Niño , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Subgrupos de Linfocitos B , Alergia e Inmunología , Linfocitos , Pancitopenia , Alergia e Inmunología , Patología , Subgrupos de Linfocitos T , Alergia e Inmunología , TelómeroRESUMEN
<p><b>OBJECTIVE</b>To explore the pathogenesis of abnormal WT1 expression in paroxysmal nocturnal hemoglobinuria (PNH).</p><p><b>METHODS</b>The expression of WT1 mRNA in CD59⁻ and CD59⁺ bone marrow mononuclear cells (BMMNC) were measured by semi-quantitative reverse transcription PCR. After WT1 gene silence by RNA interference (RNAi) technology, biological characteristics of BMMNC were investigated by flow cytometry.</p><p><b>RESULTS</b>The relative expression of WT1 mRNA in PNH CD59⁻ BMMNC (1.06 ± 0.12) was significantly higher than that in PNH CD59⁺ BMMNC (0.90 ± 0.12) and normal BMMNC (0.86 ± 0.05, P<0.05), but there was no significant difference between PNH CD59⁺ BMMNC and normal BMMNC (P>0.05). WT1 mRNA expression in PNH was positively correlated with the proportion of CD59⁻ cells (r²=0.490, P=0.016), but had no relationship with the proportion of CD59⁺ cells. After WT1 gene silence by siRNA in PNH CD59⁻ BMMNC, WT1 mRNA expression was decreased. The proportions of G0/G1 phase in PNH CD59⁻ cell blank control group and siRNA-scr transfected group were (92.73 ± 3.71)% and (93.06 ± 4.14)%, and the proportions of S phase were (6.99 ± 3.61)% and (6.73 ± 4.08)%, respectively. The proportions of G0/G1 and S phase in siRNA-WT1 transfected group was (94.46 ± 3.71)% and (5.40 ± 3.55)%, respectively. There were significant differences in the proportions of G0/G1 phase and S phase among the controls, siRNA-WT1 transfected group and siRNA-scr transfected group (P<0.05). The rate of apoptosis in siRNA-WT1 transfected group [(35.91 ± 22.36)%] was significantly higher than those in controls [(26.12 ± 17.10)%] and siRNA-scr transfected group [(27.39 ± 18.99)%] (P<0.05).</p><p><b>CONCLUSION</b>siRNA-WT1 could effectively suppress the WT1 gene expression of CD59⁻ clone in PNH patients, inhibit its proliferation, and promote its apoptosis. WT1 gene expression might contribute to PNH clone proliferation.</p>
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Adolescente , Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Apoptosis , Células de la Médula Ósea , Metabolismo , Ciclo Celular , Hemoglobinuria Paroxística , Metabolismo , Patología , Leucocitos Mononucleares , Metabolismo , Interferencia de ARN , ARN Mensajero , Genética , Proteínas WT1 , Genética , MetabolismoRESUMEN
To investigate the role of CD4+, CD25+, and CD127low regulatory T cells (Tregs) in multiple myeloma (MM). Methods:Levels of CD4+T cells and Tregs, as well as expression of CTLA-4 and apoptosis-related proteins, such as CD95, bcl-2, and Caspase3 of Tregs in peripheral blood of 30 patients with newly diagnosed cases, 27 patients under of complete remission (CR) from multiple myeloma patients, and 25 healthy adults were analyzed by flow cytometry. Results:The percentage of CD4+T cells in the untreated group was significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was significantly higher than that of the CR group and control group (P<0.05), which in ISSⅢpatients of the untreated group was significantly higher than that in I/II(P<0.05). No significant difference of CD95 expression in Tregs was observed among the three groups. The expression of CTLA-4 in Tregs from the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls (P<0.05). The expression of bcl-2 in Tregs in the untreated group was significantly higher than that of the CR group (P<0.05) and control group (P<0.01), and so was in CR group than this in controls(P<0.05). The expression of Caspase3 in Tregs from the untreated group and CR group were all significantly lower than that of the control group (P<0.05). The percentage of Tregs in CD4+T cells in the untreated group was positively correlated with the proportion of bone marrow plasma cells (P<0.05). The percentage of Tregs in CD4+T cells from 15 MM patients who received bortezamib and dexamethasone (VD) chemotherapy was negatively correlated to the ratio of plasma cell reduction after the first VD chemotherapy (r=0.735, P<0.01). Conclusion:The level of Tregs in the peripheral blood of MM patients was positively correlated with tumor burden and progression of disease, but was negatively correlated with curative effect. The increased level of Tregs was associated with their strengthened anti-apoptosis function.
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<p><b>OBJECTIVE</b>To investigate the biological characteristics of osteoblasts cultured in vitro from bone marrow (BM) of multiple myeloma (MM) patients and to explore their generation and osteogenic potential. Effects of some factors such as bortezomib and MM patient serum on the osteoblasts were observed.</p><p><b>METHODS</b>Twenty MM patients and 10 healthy donors as controls were enrolled in this study. Osteoblasts from MM patients'BM were cultured in vitro. The generation and osteogenic potential of osteoblasts from MM patients and normal subjects were compared. The changes of their osteogenic potential and biological characteristics were observed. The antigens (CD34,CD138,CD45) on osteoblasts were catalyzed by flow cytometry. The levels of IL-7 were measured by ELISA. The BMP2 mRNAs were measured by RT-PCR.</p><p><b>RESULTS</b>Osteoblasts from MM patients'BM could be cultured in vitro. The quantity of osteoblasts from MM patients (6.3±1.5) was less than normal subjects (8.2±2.6) (P<0.05). The osteoblasts cultured with MM patient serum (7.4±1.1) were less than those without patient serum (8.2±2.6) (P<0.05). Bortezomib increased those from MM patients after 6 days culture (8.9±2.1 vs 6.3±1.5) (P<0.05). Von Kossa staining showed that there were more calcium depositions in MM osteoblasts with bortezomib (8.8±1.9) than those without bortezomib (6.1±1.6) (P<0.05), but less than those from normal controls (15.8±2.2) (P<0.05). CD138, CD45, CD34 were not detected on the cultured cells. The level of IL-7 in MM patients'serum (2.07±0.71) was higher than that in normal controls (1.62±0.15) (P<0.05). The expression of BMP2 mRNA was seen in the normal osteoblasts and MM patients'osteoblasts cultured with bortezomib, but not be seen in those without bortezomib.</p><p><b>CONCLUSION</b>Osteoblasts could be cultured in vitro from MM patients' BM. The proliferation and osteogenic potential of osteoblasts from MM patients were decreased. Bortezomib was a positive regulatory of osteoblasts and MM patient serum was a negative one. They both could affect the proliferation and osteogenic potential of osteoblasts.</p>
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Adulto , Anciano , Femenino , Humanos , Masculino , Persona de Mediana Edad , Adulto Joven , Células de la Médula Ósea , Biología Celular , Proteína Morfogenética Ósea 2 , Metabolismo , Ácidos Borónicos , Farmacología , Bortezomib , Estudios de Casos y Controles , Diferenciación Celular , Mieloma Múltiple , Sangre , Osteoblastos , Biología Celular , Osteogénesis , Pirazinas , Farmacología , Suero , Células Tumorales CultivadasRESUMEN
Objective To assess the efficacy and safety of monoclonal antibody rituximab combined with cyclophosphamide (CTX) in the treatment of refractory and recurrent autoimmune hemolytic anemia.Methods Seven cases with refractory and recurrent autoimmune hemolytic anemia ( including 1 case of Evans syndrome) were recruited during January,2007 to December,2010.Treatment regimens were as follows:rituximab:375 mg/m2,1 time/week,2-6 courses; CTX:1 g,1/10 d,2-7 courses; combined with intravenous immunoglobulin (IVIG) 5 g,1 time/week,given 1 day after rituximab administration.The efficacy and safety of this regimen were assessed during follow-up.Results All the patients showed good responses (7/7).Six patients achieved complete remission (6/7) and one achieved partial remission ( 1/7 ).Average follow-up time for the patients was 27 months.All patients remained in remission during the 12-month follow-up visits.Two patients showed elevated indirect bilirubin and increased reticulocyte counts within 24 months.One patient achieved complete remission after additional rituximab therapy,and another patient remained partial remission after cyclosporine therapy.At the time of 36-month follow-up visit,the patient relapsed and was retreated with 3 courses of rituximab combined with CTX and eventually achieved partial remission.All patients tolerated the treatment well with few mild side effects.Conclusions Rituximab combined with CTX is effective and relatively safe in patients with refractory and recurrent autoimmune hemolytic anemia.Additional treatment to relapse patients about 12-24 months after drug withdrawal continues to be effective.
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Objective To investigate the expression of TET2 and DLK1 mRNA in bone marrow CD3+ T cells of patients with myelodysplastic syndrome (MDS) and their clinical significance and to explore the potential mechanism of abnormal cell-mediated immunity.Methods CD3+ T cells were sorted by magnetic activated cell-sorting system.The expressions of TET2 and DLK1 mRNA in bone marrow CD3+ T cells from 26 MDS patients and 16 healthy controls were detected by fluorescence quantitative PCR.Results The expression of TET2 mRNA in CD3+ T cells was down-regulated in the MDS patients by (0.16 ±0.15) fold compared with the controls ( P < 0.05 ).The expression of TET2 mRNA in CD3+ T cells of MDS patients was positively correlated with serum complement C3 ( r =0.404,P < 0.05 ).The expression of DLK1 mRNA in CD3+ T cells was up-regulated in the MDS patients by (1.61 ±0.88) folds compared with the controls (P<0.05).Grouped by the chromosomes,the patients with chromosome abnormalities presented significantly higher DLK1 mRNA level than those with normal chromosomes [ ( 1.45 ± 0.44 ) folds,P <0.05 ].The expression of DLK1 mRNA in CD3+ T cells of MDS patients was positively correlated with the proportion of bone marrow blasts ( r =0.343,P < 0.05 ).Conclusions The mRNA expression of TET2 in CD3+ T cells of MDS patients was decreased while the mRNA expression of DLK1 was increased,which might decline the immune surveillance function.The findings would be useful for exploring the mechanism of immune tolerance.
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The objectives of the study were to investigate the number of myeloid dendritic cells (mDC) and plasmacytoid dendritic cells (pDC) present in peripheral blood mononuclear cells (PBMC) from severe aplastic anemia (SAA) patients before and after intensive immunosuppressive therapy (IST) and to assess the expression of co-stimulatory molecules (CD80, CD86, and CD40) expressed by dendritic cells (DC) from SAA patients. The quantities of mDC and pDC and ratios of mDC to pDC in PBMC were measured in 38 SAA patients at active phase, 19 patients at recovery phase, and 17 normal controls. The surface expression of CD80, CD86, and CD40 on DCs and B lymphocytes was analyzed in 16 SAA patients and 15 normal controls. The percentages of mDC and the ratio of mDC:pDC of SAA patients at active phase increased compared to that of healthy controls [0.65% (range 0.10-2.19%) vs. 0.40% (range 0.11-1.54%), 2.64% (range 1.07-4.33%) vs. 1.56% (range 0.89-2.27), respectively (P < 0.05)]. The percentages of mDCs in recovered SAA patients decreased to 0.43% (range 0.06-0.80), and the ratio of mDC:pDC decreased to 1.78% (range 0.49-3.07). The percentages of mDC and pDC in 10 SAA patients were 0.87% (range 0.10-1.85) and 0.35% (range 0.05-0.65) before IST, which decreased to 0.24% (range 0.06-0.52) and 0.14% (range 0.01-0.28) after IST (P < 0.05). The percentages of CD86 expression on DC of SAA patients increased compared to that of healthy controls [29.84% (range 20.28-39.40) vs. 11.97% (range 0.02-24.15), respectively (P < 0.05)]. The number of mDCs increased in SAA patients, which was associated with stage of disease. The increased number of mDCs and the high expression of costimulatory molecules (CD86) on these DCs may contribute to abnormal activation of T lymphocytes in these patients and subsequent immune system-mediated bone marrow failure.
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Anemia Aplásica/metabolismo , Células Dendríticas/metabolismo , Anemia Aplásica/sangre , Anemia Aplásica/terapia , Antígeno B7-2/metabolismo , Humanos , Terapia de InmunosupresiónRESUMEN
Objectives To detect the abnormalities of CD34+ cells differentiation and bone marrow cell cycle in myelodysplastic syndrome (MDS). Methods Fifty newly diagnosed MDS ( 17 in low risk and 33 in high risk), 8 acute myeloid leukemia preceded by MDS (MDS-AML) and 25 normal controls were enrolled into this study. Their CD34+ CD38+, CD34+CD38- bone marrow cells and bone marrow cell cycle were measured with flow cytometry. Results The mean percentages of CD34+ cells in bone marrow karyocyte of high risk [ (2.29 ±2.17)% ] and MDS-AML groups [ ( 18.69 ± 17.47)% ] were significantly higher than that of control group [ ( 0.36 ± 0.49 )%, P < 0.05 ]. The mean percentages of CD34+CD38+ cells were significantly lower in low risk, high risk and MDS-AML groups [ ( 86.09 ± 7.79 )%, ( 81.68 ± 11.82)% and (82.88 ±2.60)%, respectively] than that in control group [ (92.21 ±3.85)%, P<0.05], thus the percentages of CD34+CD38- cells were significantly higher in either MDS (low risk and high risk) or MDS-AML groups [ (13.91 ±7.79)%, (18.32 ±11.82)% or (17.13 ±2.60)%, respectively] than that in control group [ (7.79 ± 3.85 )%, P < 0.05 ]. The percentages of CD+34 CD-38 cells of MDS cases correlated directly with their International Prognostic Scoring System (IPSS) (r =0.493, P =0.001 ) and WHO Adapted Prognostic Scoring System (WPSS) ( r = 0.586, P = 0.000 ) scores. The percentages of bone marrow mononuclea cells (BMMNCs) in G0/G1 phase of in low risk, high risk and MDS-AML groups [ (94.52 ±4.32)%, (96.07 ± 3.88 )% and (94.65 ± 4.55 )%, respectively ] were significantly higher than that in control group[ (88.94 ±7.30)%, P <0.01 ], thus the percentages of BMMNCs in S and G2/M phase were significantly lower in either MDS (low risk and high risk) or MDS-AML groups than that in control group (P<0.05). MDS patients with low percentages of CD34+CD38- cells presented higher therapeutic efficacy than those with high percentages of CD34+CD38- cells, while without significant differences ( P > 0.05 ) .Conclusions There are abnormalities of differentiation of CD34+ bone marrow cells and high proportion of G0/G1 cells which indicates a G1 phase arrest in MDS that might be involved in the pathogenesis of MDS. So the examination of CD34+ bone marrow cells and cell cycle might be helpful for MDS diagnosis and assessment of prognosis and therapeutic effects.