CAJ | 학술논문

骨髓移植诱导临床心脏移植后供者特异性免疫耐受方案的探讨
골수이식유도림상심장이식후공자특이성면역내수방안적탐토
A novel bone marrow transplantation strategy for donor-specific tolerance induction after heart transplantation

万方数据

中华器官移植杂志中華器官移植雜誌 중화기관이식잡지
CHINESE JOURNAL OF ORGAN TRANSPLANTATION
2011年 1期 32-35 ,共4页

郭可泉%孟旭%余元龙%韩杰%姜海明%许晓军%陆小军%贾一新%郑俊猛%张海波%李岩%郑铁%许春雷%曾文%王坚刚%崔永强%罗天戈%王珺%Ikehara Susumu

곽가천%맹욱%여원룡%한걸%강해명%허효군%륙소군%가일신%정준맹%장해파%리암%정철%허춘뢰%증문%왕견강%최영강%라천과%왕군%Ikehara Susumu

心脏移植%骨髓移植%灌流法%免疫耐受%嵌合体心髒移植%骨髓移植%灌流法%免疫耐受%嵌閤體
심장이식%골수이식%관류법%면역내수%감합체
Heart transplantation%Bone marrow transplantation%Perfusion method%Immune tolerance%Chimera

目的探讨骨髓移植诱导临床心脏移植后供者特异性免疫耐受的可行性.方法采取供心的同时采用改良"灌流法"获取供者的骨髓350 ml,经过滤及离心处理后,加入细胞冷冻保护液共80ml,分装于低温冻存袋,经程序降温,置于-80℃冰箱中保存.在常规原位心脏移植术后40 d,取冻存骨髓快速复温,穿刺受者双侧髂后上嵴,立即行骨髓腔内骨髓细胞输注(IBM-BMT),共输注单核细胞1.2×107/kg,CD34+细胞2.38×105/kg.骨髓输注前3 d行预处理,包括应用氟达拉滨、抗胸腺细胞球蛋白及全身淋巴结照射.骨髓移植后静脉应用他克莫司(Tac),维持血Tac浓度谷值在10～20μg/L;3周后改为口服Tac+吗替麦考酚酯(MMF);6周后改为环孢素A及MMF.分别于心脏移植后2、4、8和12周采集受者外周血,分别于术后4、8和12周采集受者的骨髓,应用短串联重复序列-聚合酶链反应法检测供者嵌合体.心脏移植后每周行心肌内心电图检查,每月行心肌活检1次.术后3个月,取受者及第三者外周血单核细胞,行混合淋巴细胞反应(MLR).结果心脏移植后1、2及3个月时受者的外周血及髂骨内骨髓细胞中供者来源的细胞比例分别为26.3%、19.1%、4.8%和46.3%、24.4%、7.6%.IBM-BMT后心肌内心电图监测显示心肌阻抗及R波波幅无明显变化.术后3个月行心内膜心肌活检,未见排斥反应征象.术后3个月时行超声心动图检查,提示心脏舒张、收缩功能良好.MLR提示受者对供者特异性刺激呈现低反应性,而对第三者仍保持良好的免疫活性(P＜0.01).结论采取分期骨髓移植免疫耐受诱导方案可安全、有效地建立嵌合体,成功诱导心脏移植后供者特异性免疫耐受,但远期效果有待进一步研究.
목적 탐토골수이식유도림상심장이식후공자특이성면역내수적가행성.방법 채취공심적동시채용개량"관류법"획취공자적골수350 ml,경과려급리심처리후,가입세포냉동보호액공80ml,분장우저온동존대,경정서강온,치우-80℃빙상중보존.재상규원위심장이식술후40 d,취동존골수쾌속복온,천자수자쌍측가후상척,립즉행골수강내골수세포수주(IBM-BMT),공수주단핵세포1.2×107/kg,CD34+세포2.38×105/kg.골수수주전3 d행예처리,포괄응용불체랍빈、항흉선세포구단백급전신림파결조사.골수이식후정맥응용타극막사(Tac),유지혈Tac농도곡치재10～20μg/L;3주후개위구복Tac+마체맥고분지(MMF);6주후개위배포소A급MMF.분별우심장이식후2、4、8화12주채집수자외주혈,분별우술후4、8화12주채집수자적골수,응용단천련중복서렬-취합매련반응법검측공자감합체.심장이식후매주행심기내심전도검사,매월행심기활검1차.술후3개월,취수자급제삼자외주혈단핵세포,행혼합림파세포반응(MLR).결과심장이식후1、2급3개월시수자적외주혈급가골내골수세포중공자래원적세포비례분별위26.3%、19.1%、4.8%화46.3%、24.4%、7.6%.IBM-BMT후심기내심전도감측현시심기조항급R파파폭무명현변화.술후3개월행심내막심기활검,미견배척반응정상.술후3개월시행초성심동도검사,제시심장서장、수축공능량호.MLR제시수자대공자특이성자격정현저반응성,이대제삼자잉보지량호적면역활성(P＜0.01).결론 채취분기골수이식면역내수유도방안가안전、유효지건립감합체,성공유도심장이식후공자특이성면역내수,단원기효과유대진일보연구.
Objective To investigate a new strategy of bone marrow transplantation (BMT) for donor-specific tolerance induction after heart transplantation. Methods Donor bone marrow cells (BMCs)were harvested simultaneously with donor cardiac graft using modified perfusion method (PM) ,then stored in a -80 ℃ refrigerator after filtration and centrifugation. Whole BMCs (IBM-BMT) (monocytes 1.2 ×107/kg,CD34+ cells 2.38× 105/kg) in host iliac bones were injected into the bone marrow cavity 40 days after heart transplantation. Preconditoning regimens that consisted of fludarabine, antithymoctye globin and total lymphoid irradiation were performed 3 days before BMT. Tacrolimus (Tac) was administrated intravenously after BMT or orally in conjunction with mycophenolate mofetil (MMF) 3 weeks later.Cyclosporine and MMF were orally administrated 6 weeks later. Donor chimerism was detected using short tandem repeats-polymerase chain reaction in monocytes from peripheral blood at the 2nd,4th, 8th or 12th week after BMT or BMCs at the 4th, 8th or 12th week after BMT. Intramyocardium electrocardiography examination or endomyocardial biopsy was performed weekly or monthly respectively. Mixed lymphocyte reactions (MLR) were performed 3 months after BMT. Results Donor chimerism in monocytes in peripheral blood or BMCs in iliac bones measured at the 1 st,2nd and 3rd month after BMT was 26.3%, 19.1%,4.8% ,and 46.3%, 24.4%, 7.6%, respectively. After 3-month follow-up, there was no rejection confirmed by endomyocardial biopsy or intramyocardium electrocardiography. Echocardiography revealed that the diastolic and systolic function of the cardiac graft was maintained well 3 months after BMT. MLR revealed donor-specific hyporesponsiveness while immunocompetence was preserved to third-party antigens. Conclusion These findings indicate that the two-stage BMT strategy is a safe and feasible method for the induction of donor-specific tolerance via stable mixed chimerism and needs to be further confirmed after a long-term observation.