CAJ | 학술논문

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HLA-B27-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。目的：觀察免疫相關性血細胞減少綜閤徵（ Immune related hematocytopenia syndrome ，IRHS）患者骨髓造血微環境免疫分子錶達狀態，對骨髓單箇覈細胞（ BMMNC）耑粒酶活性（ TA）的影響，探討對造血細胞破壞的免疫損傷機製及臨床意義。方法：①耑粒重複序列擴增-酶聯免疫吸附法（ TRAP-PCR-ELISA）檢測366例IRHS患者治療前後BMMNC的TA。②免疫化學染色和免疫熒光染色，觀察患者骨髓細胞和造血微環境中HLA-DR、抗人IgG、FcγⅡ受體（ FcγⅡR）、甘露糖受體（ MR）、白細胞介素17A（IL-17A）及受體錶達。③FACS技術檢測患者外週血淋巴細胞CD3＋CD4＋T細胞、CD3＋CD8＋T細胞、CD3-CD16＋／CD56＋NK細胞和CD19＋B細胞比例。60例健康查體者外週血淋巴細胞亞群作為正常對照，30例缺鐵性貧血患者骨髓作為疾病對照。結果：①IRHS患者治療前耑粒酶活性為0.2617±0.0216，高于疾病對照組（0.0616±0.0313， P＜0.01）；其中HLA-B27＋組高于HLA-B27-組（0.3013±0.0206比0.1923±0.0129，P＜0.05），HLA-B27＋IgG升高的重度IRP患者顯著高于HLA-B27-IgG升高組（0.4016±0.0172比0.2211±0.0110，P＜0.01）。②HLA-B27＋IgG＋組骨髓免疫細胞和造血微環境呈現細胞免疫活化和體液免疫均紊亂狀態，高錶達HLA-DR、FcγⅡR、IL-17A、IL-17RA和MR等免疫分子，外週血CD3＋CD4＋T細胞、CD3＋CD8＋T細胞和CD3-CD16＋／CD56＋NK和CD19＋B細胞比例呈不同程度升高，造血破壞呈現多樣性。③而HLA-B27-IgG＋組骨髓免疫細胞和造血微環境呈現體液免疫活化優勢狀態，高錶達FcγⅡR，IL-17A等炎性因子不錶達或弱錶達，外週血僅CD19＋B細胞比例升高，骨髓造血破壞方式主要為抗體依賴性細胞毒性（ ADCC）效應。經糖皮質激素聯閤環孢素A等藥物治療後，患者血象恢複時BMMC的耑粒酶重新失活，TA為0。結論：IRHS骨髓單箇覈細胞耑粒酶活化，與患者骨髓造血微環境免疫狀態和病變血細胞病理性損害程度密切相關。病毒感染和細胞免疫活化，多種炎性因子參與IRHS造血免疫損傷，是TA增彊的重要因素。
목적：관찰면역상관성혈세포감소종합정（ Immune related hematocytopenia syndrome ，IRHS）환자골수조혈미배경면역분자표체상태，대골수단개핵세포（ BMMNC）단립매활성（ TA）적영향，탐토대조혈세포파배적면역손상궤제급림상의의。방법：①단립중복서렬확증-매련면역흡부법（ TRAP-PCR-ELISA）검측366례IRHS환자치료전후BMMNC적TA。②면역화학염색화면역형광염색，관찰환자골수세포화조혈미배경중HLA-DR、항인IgG、FcγⅡ수체（ FcγⅡR）、감로당수체（ MR）、백세포개소17A（IL-17A）급수체표체。③FACS기술검측환자외주혈림파세포CD3＋CD4＋T세포、CD3＋CD8＋T세포、CD3-CD16＋／CD56＋NK세포화CD19＋B세포비례。60례건강사체자외주혈림파세포아군작위정상대조，30례결철성빈혈환자골수작위질병대조。결과：①IRHS환자치료전단립매활성위0.2617±0.0216，고우질병대조조（0.0616±0.0313， P＜0.01）；기중HLA-B27＋조고우HLA-B27-조（0.3013±0.0206비0.1923±0.0129，P＜0.05），HLA-B27＋IgG승고적중도IRP환자현저고우HLA-B27-IgG승고조（0.4016±0.0172비0.2211±0.0110，P＜0.01）。②HLA-B27＋IgG＋조골수면역세포화조혈미배경정현세포면역활화화체액면역균문란상태，고표체HLA-DR、FcγⅡR、IL-17A、IL-17RA화MR등면역분자，외주혈CD3＋CD4＋T세포、CD3＋CD8＋T세포화CD3-CD16＋／CD56＋NK화CD19＋B세포비례정불동정도승고，조혈파배정현다양성。③이HLA-B27-IgG＋조골수면역세포화조혈미배경정현체액면역활화우세상태，고표체FcγⅡR，IL-17A등염성인자불표체혹약표체，외주혈부CD19＋B세포비례승고，골수조혈파배방식주요위항체의뢰성세포독성（ ADCC）효응。경당피질격소연합배포소A등약물치료후，환자혈상회복시BMMC적단립매중신실활，TA위0。결론：IRHS골수단개핵세포단립매활화，여환자골수조혈미배경면역상태화병변혈세포병이성손해정도밀절상관。병독감염화세포면역활화，다충염성인자삼여IRHS조혈면역손상，시TA증강적중요인소。
Objective:To study the effect of immunological molecules expressive state upon the telomerase activation ( TA) of bone marrow mononuclear cells ( BMMNC ) in the hemopoietic microenvironment of patients with immune related hematocytopenia ( IRHS) ,and to explore the immunologic mechanism as well as the clinical significance of hematoclasis in marrow of IRHS patients .Methods:①TRAP-PCR-ELISA method was performed to detect the TA of BMMNC in marrow of 366 IRHS patients before and after therapy.②The molecules HLA-DR,anti-hunman IgG,FcγⅡR,mannose receptor ( MR),IL-17A and its receptor ( IL-17AR) were analysed by immunochemistry and immunofluorescence staining .③The flow cytometric ( FCM) was used to analyse the proportion of CD3+CD4+T cells as well as CD3+CD8+T cells ,CD3-CD19+B cells and CD3-CD16/56+NK cells in peripheral blood lymphocyte.60 cases of health examination were selected as the control group , and 30 cases hypoferric anemia patients were selected as disease control.The differences between patient group and control group were analysed with statistic method .Immunochemistry and immunofluo-rescence staining were performed to in situ analyze the activation-characteristics of immunocyte in bone marrow slides of IRHS ,and the dependablity of cellular immunologic injury was also checked.Results: ①The levels of TA was 0.261 7 ±0.021 6 before treatment , higher than the disease control group (0.061 6±0.031 3 ,P<0.01).Among of them HLA-B27+patients were higher than HLA-B27-patients (0.301 3±0.020 6 vs.0.192 3±0.012 9,P<0.05).Serious IRP patients with HLA-B27+IgG+were obviously higher than HLA-B27-IgG+patients (0.401 6±0.017 2 vs.0.221 1±0.011 0,P<0.01).②In marrow of HLA-B27+IgG+patients,both cell immunity and humoral immunity were in disorder in the hemopoietic microenvironment ,and immonocyte in marrow expressed HLA-DR, FcγⅡR,IL-17A,IL-17RA and MR,and Th,Ts,B cell and NK cell in peripheral blood increased in different degree ,inducing the in-flammation of haemocyte and lead to destruction.③Humoral immunity was in the dominant level in morrow;humoral immunity of HLA-B27-IgG+patients,immonocyte expressed FcγⅡR in high level,but IL-17A was seldom expressed,only CD19+B cell was increased slightly ,the antibody dependent cellular cytotoxicity ( ADCC) was the main mode of destruction.After therapy glucocorticoids associated with ciclosporin A ,the TA level of BMMNC decreased to 0 with devitalization.Conclusion: The telomerase activation of bone marrow mononuclear cells in IRHS is related with the immune state of hemopoietic microenvironment and the pathologic lesion degree of hema -topoietic cell in marrow.It is viral infection and immunological activation as well as a variety of inflammatory factors play a part in the immunologic injury that might be an important factor of the enhancement in TA.