Objective To observe the expression of inlfammatory molecules in bone marrow immune cells of patients with immune-related hematocytopenia (IRH), and to investigate the immune mechanism and clinical signiifcance of the...Objective To observe the expression of inlfammatory molecules in bone marrow immune cells of patients with immune-related hematocytopenia (IRH), and to investigate the immune mechanism and clinical signiifcance of the disease. Methods Total of 36 IRH patients were selected as observation group and 30 healthy people were taken as control group. Serum cytokines levels, activity of immunocytes and expression of HLA-DR were detected. Immune lfuorescence was applied to observe the expression state of immunologic molecules and cytokines in IRH patients. Results Serum cytokines were elevated in various degrees in observation group. Compared with the control group, the cytokines levels were significantly higher (P < 0.05). After treatement with immunosuppressive drugs, the serum levels of cytokines in observation group reduced to a level close to the control group. HLA-DR were upregulated in activated tissue basophils, eosinophils, dendritic cells (DC) and macrophages of bone marrow in IRH patients, and POX activity in these immunocytes of IRH was higher than that of the control group. Immune molecules were highly expressed in eosinophils, DC and macrophages. Conclusions It is demonstrated that antibodies or self-reactive lymphocytes were produced in IRH marrow, which would cause lesions of hemocytes, and lead to pathological process ifnally. Structure of hematopoietic cells mutated and these cells might be acted as target cells of immunocytes in the pathological process. Immunocytes could secrete inlfammatory factors and lead to immunologic injury of hemocyte.展开更多
Background Immune-related hematocytopenia （IRH） is considered to be related with the production of autoantibody, as well as the activation of humoral immunity which is stimulated by B lymphocyte. This study aimed to...Background Immune-related hematocytopenia （IRH） is considered to be related with the production of autoantibody, as well as the activation of humoral immunity which is stimulated by B lymphocyte. This study aimed to observe the levels of various cytokines in the blood serum and the in situ active state of macrophage （Me） in the medullary hematopoietic microenvironment of IRH patients, and to probe into the immune mechanism and clinical significance of Me in hematopoietic cell injury. Methods ELISA is used to detect the IL-4, IL-6, IL-12, IL-17, and IFN-y levels in the peripheral blood serum of 376 patients in pre- and post-therapy. Cytochemistry and cell immunochemistry methods are used to observe the peroxidase （POX）, nonspecific esterase （NSE）, hemosiderin granules, and HLA-DR activity of Me in the bone marrow of patients. Immunofluorescence is used to observe the expression of hemocyte antihuman globulin IgG antibody, lymphocytes CD4 molecule, Me membrane Fcyllreceptor （FcyllR）, mannitose receptor （MR）, IFN-y, ICAM-1, IL-12, and IL-17A and the formation mechanism of antibody-dependent cell-mediated cytotoxicity （ADCC） hematopoietic cell islands （HI） in the medullary hematopoietic microenvironment of patients. Glucocorticoid is used for treatment on the basis of anti-infection therapy, and gamma globulin stoss therapy is used for the appearance of ADCC-type HI or serious Me bloodthirsty phenomenon; if necessary, association of Cyclosporine A （CsA） should be used and chalybeate should be supplemented. Results In the patient group, the levels of IL-4, IL-6, IL-12, IL-17, and IFN-y were increased. After treatment, the cytokine levels gradually became normal. The activated Me in the marrow highly expressed NSE and POX, and Me swallowed more hemosiderin particles, but the iron in the cytoplasm of immature erythrocytes decreased. The activated Me expressed HLA-DR, MR, ICAM-1, IFN-y, and IL-12. For patients with humoral immunity activation and bacterial infection, Me weakly expressed IL-17A but highly expressed FcyIIR, and the phenomenon that ADCC-type HI broke pathological blood corpuscles often occurred; for the cellular immune activation along with virus infection, the white blood count （WBC） significantly reduced, Me weakly expressed FcyIIR, secretory highly expressed IL-17A, and the phenomena that Me adhered to, captured and swallowed blood cell often occurred. After four weeks of anti-infective and immunosuppressive therapy, nuclear apoptosis of Me occurred in the bone marrow of patients, HI and bloodthirsty phenomenon disappeared, and the peripheral blood picture started to improve. Conclusions Me is an important antigen presenting cell in the IRH marrow for hematopoiesis destruction and an immune effector cell of hematopoietic injury; infection can promote the activation of Me, upregulate the impression of immune molecule and receptors, form ADCC HI. aeGravate hematoBoietic iniurv, and accelerate the destruction on hematoDoietic cell.展开更多
目的:观察免疫相关性血细胞减少综合征(Immune related hematocytopenia syndrome,IRHS)患者骨髓造血微环境免疫分子表达状态,对骨髓单个核细胞(BMMNC)端粒酶活性(TA)的影响,探讨对造血细胞破坏的免疫损伤机制及临床意义。方法:①端粒...目的:观察免疫相关性血细胞减少综合征(Immune related hematocytopenia syndrome,IRHS)患者骨髓造血微环境免疫分子表达状态,对骨髓单个核细胞(BMMNC)端粒酶活性(TA)的影响,探讨对造血细胞破坏的免疫损伤机制及临床意义。方法:①端粒重复序列扩增-酶联免疫吸附法(TRAP-PCR-ELISA)检测366例IRHS患者治疗前后BMMNC的TA。②免疫化学染色和免疫荧光染色,观察患者骨髓细胞和造血微环境中HLA-DR、抗人IgG、FcγⅡ受体(FcγⅡR)、甘露糖受体(MR)、白细胞介素17A(IL-17A)及受体表达。③FACS技术检测患者外周血淋巴细胞CD3+CD4+T细胞、CD3+CD8+T细胞、CD3-CD16+/CD56+NK细胞和CD19+B细胞比例。60例健康查体者外周血淋巴细胞亚群作为正常对照,30例缺铁性贫血患者骨髓作为疾病对照。结果:①IRHS患者治疗前端粒酶活性为0.2617±0.0216,高于疾病对照组(0.0616±0.0313,P<0.01);其中HLA-B27+组高于HLA-B27-组(0.3013±0.0206比0.1923±0.0129,P<0.05),HLA-B27+IgG升高的重度IRP患者显著高于HLA-B27-IgG升高组(0.4016±0.0172比0.2211±0.0110,P<0.01)。②HLA-B27+IgG+组骨髓免疫细胞和造血微环境呈现细胞免疫活化和体液免疫均紊乱状态,高表达HLA-DR、FcγⅡR、IL-17A、IL-17RA和MR等免疫分子,外周血CD3+CD4+T细胞、CD3+CD8+T细胞和CD3-CD16+/CD56+NK和CD19+B细胞比例呈不同程度升高,造血破坏呈现多样性。③而HLAB27-IgG+组骨髓免疫细胞和造血微环境呈现体液免疫活化优势状态,高表达FcγⅡR,IL-17A等炎性因子不表达或弱表达,外周血仅CD19+B细胞比例升高,骨髓造血破坏方式主要为抗体依赖性细胞毒性(ADCC)效应。经糖皮质激素联合环孢素A等药物治疗后,患者血象恢复时BMMC的端粒酶重新失活,TA为0。结论:IRHS骨髓单个核细胞端粒酶活化,与患者骨髓造血微环境免疫状态和病变血细胞病理性损害程度密切相关。病毒感染和细胞免疫活化,多种炎性因子参与IRHS造血免疫损伤,是TA增强的重要因素。展开更多
文摘Objective To observe the expression of inlfammatory molecules in bone marrow immune cells of patients with immune-related hematocytopenia (IRH), and to investigate the immune mechanism and clinical signiifcance of the disease. Methods Total of 36 IRH patients were selected as observation group and 30 healthy people were taken as control group. Serum cytokines levels, activity of immunocytes and expression of HLA-DR were detected. Immune lfuorescence was applied to observe the expression state of immunologic molecules and cytokines in IRH patients. Results Serum cytokines were elevated in various degrees in observation group. Compared with the control group, the cytokines levels were significantly higher (P < 0.05). After treatement with immunosuppressive drugs, the serum levels of cytokines in observation group reduced to a level close to the control group. HLA-DR were upregulated in activated tissue basophils, eosinophils, dendritic cells (DC) and macrophages of bone marrow in IRH patients, and POX activity in these immunocytes of IRH was higher than that of the control group. Immune molecules were highly expressed in eosinophils, DC and macrophages. Conclusions It is demonstrated that antibodies or self-reactive lymphocytes were produced in IRH marrow, which would cause lesions of hemocytes, and lead to pathological process ifnally. Structure of hematopoietic cells mutated and these cells might be acted as target cells of immunocytes in the pathological process. Immunocytes could secrete inlfammatory factors and lead to immunologic injury of hemocyte.
文摘Background Immune-related hematocytopenia （IRH） is considered to be related with the production of autoantibody, as well as the activation of humoral immunity which is stimulated by B lymphocyte. This study aimed to observe the levels of various cytokines in the blood serum and the in situ active state of macrophage （Me） in the medullary hematopoietic microenvironment of IRH patients, and to probe into the immune mechanism and clinical significance of Me in hematopoietic cell injury. Methods ELISA is used to detect the IL-4, IL-6, IL-12, IL-17, and IFN-y levels in the peripheral blood serum of 376 patients in pre- and post-therapy. Cytochemistry and cell immunochemistry methods are used to observe the peroxidase （POX）, nonspecific esterase （NSE）, hemosiderin granules, and HLA-DR activity of Me in the bone marrow of patients. Immunofluorescence is used to observe the expression of hemocyte antihuman globulin IgG antibody, lymphocytes CD4 molecule, Me membrane Fcyllreceptor （FcyllR）, mannitose receptor （MR）, IFN-y, ICAM-1, IL-12, and IL-17A and the formation mechanism of antibody-dependent cell-mediated cytotoxicity （ADCC） hematopoietic cell islands （HI） in the medullary hematopoietic microenvironment of patients. Glucocorticoid is used for treatment on the basis of anti-infection therapy, and gamma globulin stoss therapy is used for the appearance of ADCC-type HI or serious Me bloodthirsty phenomenon; if necessary, association of Cyclosporine A （CsA） should be used and chalybeate should be supplemented. Results In the patient group, the levels of IL-4, IL-6, IL-12, IL-17, and IFN-y were increased. After treatment, the cytokine levels gradually became normal. The activated Me in the marrow highly expressed NSE and POX, and Me swallowed more hemosiderin particles, but the iron in the cytoplasm of immature erythrocytes decreased. The activated Me expressed HLA-DR, MR, ICAM-1, IFN-y, and IL-12. For patients with humoral immunity activation and bacterial infection, Me weakly expressed IL-17A but highly expressed FcyIIR, and the phenomenon that ADCC-type HI broke pathological blood corpuscles often occurred; for the cellular immune activation along with virus infection, the white blood count （WBC） significantly reduced, Me weakly expressed FcyIIR, secretory highly expressed IL-17A, and the phenomena that Me adhered to, captured and swallowed blood cell often occurred. After four weeks of anti-infective and immunosuppressive therapy, nuclear apoptosis of Me occurred in the bone marrow of patients, HI and bloodthirsty phenomenon disappeared, and the peripheral blood picture started to improve. Conclusions Me is an important antigen presenting cell in the IRH marrow for hematopoiesis destruction and an immune effector cell of hematopoietic injury; infection can promote the activation of Me, upregulate the impression of immune molecule and receptors, form ADCC HI. aeGravate hematoBoietic iniurv, and accelerate the destruction on hematoDoietic cell.
文摘目的:观察免疫相关性血细胞减少综合征(Immune related hematocytopenia syndrome,IRHS)患者骨髓造血微环境免疫分子表达状态,对骨髓单个核细胞(BMMNC)端粒酶活性(TA)的影响,探讨对造血细胞破坏的免疫损伤机制及临床意义。方法:①端粒重复序列扩增-酶联免疫吸附法(TRAP-PCR-ELISA)检测366例IRHS患者治疗前后BMMNC的TA。②免疫化学染色和免疫荧光染色,观察患者骨髓细胞和造血微环境中HLA-DR、抗人IgG、FcγⅡ受体(FcγⅡR)、甘露糖受体(MR)、白细胞介素17A(IL-17A)及受体表达。③FACS技术检测患者外周血淋巴细胞CD3+CD4+T细胞、CD3+CD8+T细胞、CD3-CD16+/CD56+NK细胞和CD19+B细胞比例。60例健康查体者外周血淋巴细胞亚群作为正常对照,30例缺铁性贫血患者骨髓作为疾病对照。结果:①IRHS患者治疗前端粒酶活性为0.2617±0.0216,高于疾病对照组(0.0616±0.0313,P<0.01);其中HLA-B27+组高于HLA-B27-组(0.3013±0.0206比0.1923±0.0129,P<0.05),HLA-B27+IgG升高的重度IRP患者显著高于HLA-B27-IgG升高组(0.4016±0.0172比0.2211±0.0110,P<0.01)。②HLA-B27+IgG+组骨髓免疫细胞和造血微环境呈现细胞免疫活化和体液免疫均紊乱状态,高表达HLA-DR、FcγⅡR、IL-17A、IL-17RA和MR等免疫分子,外周血CD3+CD4+T细胞、CD3+CD8+T细胞和CD3-CD16+/CD56+NK和CD19+B细胞比例呈不同程度升高,造血破坏呈现多样性。③而HLAB27-IgG+组骨髓免疫细胞和造血微环境呈现体液免疫活化优势状态,高表达FcγⅡR,IL-17A等炎性因子不表达或弱表达,外周血仅CD19+B细胞比例升高,骨髓造血破坏方式主要为抗体依赖性细胞毒性(ADCC)效应。经糖皮质激素联合环孢素A等药物治疗后,患者血象恢复时BMMC的端粒酶重新失活,TA为0。结论:IRHS骨髓单个核细胞端粒酶活化,与患者骨髓造血微环境免疫状态和病变血细胞病理性损害程度密切相关。病毒感染和细胞免疫活化,多种炎性因子参与IRHS造血免疫损伤,是TA增强的重要因素。
