非清髓性骨髓移植对移植物抗宿主病的影响

目的探讨非清髓性预处理方案对大鼠异基因骨髓移植模型移植物抗宿主病(GVHD)的影响.方法Wistar大鼠为供鼠,SD大鼠为受鼠,受鼠分两组,分别予以不同的预处理方案,A组受鼠自移植前4～1 d,腹腔注射氟达拉宾(1 mg/kg),移植前4 h接受2Gy的全身照射;B组受鼠仅在移植当天分别接受6 Gy的全身照射.各实验组受鼠均在移植当天经尾静脉输注供鼠骨髓细胞3.8×107;对照组大鼠输注等量的生理盐水,观察各组大鼠GVHD反应.结果移植前接受氟达拉宾和2 Gy全身照射预处理的大鼠生存期较长,且 GVHD 临床和病理评分也较低.结论非清髓性骨髓移植预处理方案可有效预防GVHD,拓宽临床进行异基因骨髓移植的适用范围.     

非清髓性骨髓移植对移植物抗宿主病的影响

目的 探讨非清髓性预处理方案对大鼠异基因骨髓移值模型移植物抗宿主病(GVHD)的影响。方法 Wistar大鼠为供鼠,SD大鼠为受鼠,受鼠分两组,分别予以不同的预处理方案,A组受鼠自移植前4-1d,腹腔注射氟达拉宾(1mg/kg),移植前4h接受2Gy的全身照射;B组受鼠仅在移植当天分别接受6Gy的全身照射。各实验组受鼠均在移植当天经尾静脉输注供鼠骨髓细胞3.8×107;对照组大鼠输注等量的生理盐水,观察各组大鼠GVHD反应。结果 移植前接受氟达拉宾和2Gy全身照射预处理的大鼠生存期较长,且GVHD临床和病理评分也较低。结论 非清髓性骨髓移植预处理方案可有效预防GVHD,拓宽临床进行异基因骨髓移植的适用范围。     

自然杀伤细胞对小鼠异基因骨髓移植中移植物抗宿主病效应的影响

目的:研究自然杀伤细胞(NK)在异基因骨髓移植中对移植物抗宿主病(GVHD)的影响.方法:以近交系小鼠C57BL/6(H-2b)为供鼠、BALB/c(H-2d)为受鼠,在移植物中增加供者的外周T细胞和(或)NK细胞进行异基因骨髓移植,根据临床表现和病理检查,比较不同移植组的存活率和GVHD发生情况.结果:增加1×106供鼠NK细胞的移植组中40%小鼠出现GVHD,增加2×106供鼠NK细胞的移植组中仅30%出现GVHD;与单纯异基因骨髓移植细(80%急性GVHD、20%慢性GVHD)和骨髓 T细胞组(100%急性GVHD)比较,GVHD发生率显著下降(P＜0.01),30、60和120 d存活率显著增高(P＜0.01).结论:在小鼠异基因骨髓移植中,同种异基因反应性NK细胞可抑制GVHD效应,延长存活期.     

非亲缘异基因骨髓移植后并发急性移植物抗宿主病的诊治

移植物抗宿主病(ＧＶＨＤ)是异基因造血干细胞移植的主要障碍。我院近１年完成的１０例非亲缘异基因骨髓移植中 ,１例发生超急性移植物抗宿主病 (ａＧＶＨＤ) ,经抢救好转 ,现报道如下。１．供受体情况 :患者女性 ,１２岁 ,山东省临沂市学生。７年前发现血白细胞数增高 ,２年前血白细胞数升至１３０×１０９／Ｌ,当地骨髓涂片检查确诊为慢性髓细胞白血病 (ＣＭＬ)慢性期 ,予羟基脲治疗。移植前血象 :白细胞数７．５×１０９／Ｌ,血红蛋白量１０９ｇ／Ｌ,血小板数２９６×１０９／Ｌ;骨髓象 :有核细胞增多 ,粒系增生明显活跃 ,以中幼粒以下阶…     

异种骨髓移植中移植物抗宿主病的实验研究

目的 :探讨克服异种骨髓移植间强烈的移植物抗宿主病的方法。方法 :实验分两步 :第一步 ,SD大鼠亚致死剂量 5 .5Gy全身照射后 ,4h内经尾静脉输入正常BALB/c小鼠骨髓细胞 8× 10 7。 2d后予腹腔注射Cy15 0mg/kg。诱导形成嵌合体大鼠 ,使其对BALB/c小鼠产生特异性免疫耐受。第二步 ,BALB/c小鼠接受致死剂量 9.0Gy全身照射 ,随机三分组。照射后 4h内经尾静脉注射骨髓进行移植。A组输注正常SD大鼠的骨髓细胞 4× 10 7;B组输注正常SD大鼠的骨髓细胞 4× 10 7和脾细胞 2× 10 7;C组输注嵌合体SD大鼠的骨髓细胞 4× 10 7和脾细胞 2× 10 7;比较GVHD发生情况。结果 :A组有 2只小鼠死于感染和放射损伤 ,所有对象均未观察到明显GVHD表现。B组平均存活时间为 (9.0± 1.3)d ,且死前均出现不同程度的典型GVHD表现 ,如皮毛脏乱、消瘦、弓背体位、腹泻 ,甚至血便等 ;肝实质内有灶性淋巴细胞及多形细胞浸润 ,并可见局灶性坏死 ;肠绒毛部分或大部分脱落 ,粘膜坏死、有大量炎性细胞浸润 ,符合GVHD病理改变。而C组除 2只小鼠分别于 18d、31d死亡外 ,余均存活超过 6 0d ,与B组有显著差异 ,且死亡率最低。结论 :有受者嵌合体的供者 ,在进行异种骨髓移植后有助于克服异种移植物抗宿主病。     

单倍体相合骨髓移植新技术治疗难治复发白血病

国内单生子女政策及无一定规模的骨髓库，使多数需要造血干细胞移植的白血病病人找到HLA匹配相关与非相关供者非常困难，但几乎所有人均有单倍体相合相关供者，它为造血干细胞移植治疗奠定了基础。80年代国外开始单倍型相关造血干细胞移植，由于重症移植物抗宿主病(GVHD)，结果失败。去除植入物中T细胞后，可控制GVHD发生，但     

树突状细胞亚群比例对小鼠异基因骨髓移植后急性移植物抗宿主病的影响

目的 探讨树突状细胞(Dendridc cell，DC)亚群比例对小鼠异基因骨髓移植(allo-BMT)后急性移植物抗宿主病(aGVHD)的影响。方法用主要组织相容性抗原(MHc)完全不合的近交系小鼠建立小鼠allo-BMT的模型，随机分为三组，分别在移植前、后和不对供鼠使用重组小鼠粒系集落刺激因子rmG-CSF，移植后均采用环孢素A(CsA) 甲氨蝶呤(MTX)方案预防aGVHD，观察三组小鼠移植前后外周血中DC亚群的比例及变化以及aGVHD的情况。结果 移植前后使用rhG-CSF均可以使DC亚群发生改变。使DC2的比例增加，移植前使用更明显；处理组移植小鼠aGVHD严重程度明显比不使用rmG-CSF组弱。移植前使用aGVHD表现最轻。结论 移植前后使用rmG-CSF可以使小鼠DC亚群比例发生改变，使DC2比例增高，并可使减轻移植小鼠aGVHD的症状。     

异种骨髓移植后1例慢性移植物抗宿主病动物模型的观察

目的 探讨异种造血嵌合体模型中慢性移植物抗宿主病（graft-versus-host disease,GVHD）发生的特点及机制。方法 本研究对40余只异种造血嵌合体模型中出现的一只慢性GVHD实验兔行血常规、肝功、骨髓涂片检查，对其肝、肺、小肠、皮肤等脏器行常规病理、胶原纤维染色等检查，并同时用PCR测其外周血中供体细胞嵌合情况。结果 该实验兔外周全血细胞减少，PCR检查未发现供体细胞，骨髓轻度增生不良；病理结果显示其病变仍以皮肤为重但均较同种GVHD轻，其病变侵犯部位与同种GVHD无明显区别。结论 异种骨髓移植的慢性GVHD出现时间晚，较罕见，临床表现不如同种慢性GVHD典型，病变较同种GVHD轻；其病变侵犯器官与同种GVHD无明显区别。     

造血干细胞移植后的移植物抗宿主病

本院14例异基因骨髓移植中发生急性移植物抗宿主病7例(50％),其中Ⅳ级2例(14.3％)。Ⅰ～Ⅱ级5例;慢性移植物抗宿主病2例(14.3％)。9例胎肝移植中1例发生移植物抗宿主病Ⅰ级。1例自体骨髓移植后因输未经照射的异体血引起急性移植物抗宿主病。收集1988年至1991年12月国内文献报告异基因骨髓移植57例,发生急性移植物抗宿主病44例(77.19％),Ⅲ级以上12例(20.05％),慢性移植物抗宿主病14例(24.56％)。初步分析我国移植物抗宿主病的发病率与欧美早期报告近似。本文结合国内外文献报告介绍了移植物抗宿主病的防治措施。     

大鼠异基因骨髓移植急性移植物抗宿主病模型的建立

目的建立较稳定的异基因骨髓移植急性移植物抗宿主病动物模型,为异基因骨髓移植后的急性移植物抗宿主病(aGVHD)的相关研究提供实验参照。方法以雄性SD大鼠为供鼠,雌性Wistar大鼠为受鼠,受体大鼠随机分成A、B、C、D、E 5组,移植当天所有受鼠均接受8.5 GY的全身照射(TBI),于照射后4~6 h内,A组回输等量培养液,B组经尾静脉输注供鼠骨髓细胞(2×108个/kg),C、D、E组分别回输供鼠骨髓细胞(2×108个/kg) 不同比例的脾细胞。观察各组大鼠生存期、外周白细胞计数、及有无aGVHD的临床及病理表现。结果A组大鼠于15d内全部死亡,外周血白细胞计数明显减低,骨髓病理示造血组织减少,提示死于造血衰竭。B、C、D、E组大鼠外周血白细胞计数均有明显恢复,B组大鼠8只存活超过50 d,C、D、E组大鼠均于50 d观察期内死亡,并有aGVHD的临床表现及病理表现,但C组大鼠aGVHD的程度较轻且时间不集中,其中D、E组大鼠可于相对集中的时间内观察到典型aGVHD临床及病理。结论TBI预处理的方式是可行的,单纯输入异基因骨髓细胞不能引起明显的aGVHD,骨髓细胞与脾细胞1∶1及1∶1.5混合组均可作为异基因骨髓移植后理想的aGVHD动物模型。     

Effect of NK cells on GVHD in H-2 haploidentical bone marrow transplantation in mice

Objective To study the effect of natural killer (NK) cells on graft-versus-host disease (GVHD) after H-2 haploidentical bone marrow transplantation (BMT) in mice. Methods Murine model of H-2 haploidentical BMT was established by using Balb/c (H-2d) mouse as recipient, and Balb/c(H-2d)×C57BL/6 (H-2b) (H-2d/b) mouse as donor. Lethally irradiated Balb/c (H-2d) mice were transplanted with the bone marrow cells from Balb/c(H-2d)×C57BL/6(H-2b) (H-2d/b) mice containing donor spleen cells and/or NK cells. GVHD and survival rates were studied by observation of clinical manifestations and pathological changes. ResultsIn the group of bone marrow spleen cells, GVHD was induced in 90% mice; but in the group plus with low amount of NK cells,GVHD was induced in 20% mice; and in the group transplanted with high amount of NK cells, GVHD was induced only in 10% mice. Compared to the group transplanted only with BM plus spleen cells, the incidences of GVHD in the latter two groups decreased significantly (P＜0.01) and the survival rates at different periods of 15, 30, 45 and 60 days increased obviously (P＜0.01 ). Conclusion In mouse H-2 haploidentical BMT, alloreactive NK cells can reduce the incidence of GVHD and increase the survival rate.     

NK细胞对小鼠H-2单倍体相合骨髓移植后GVHD及造血重建的影响

目的:研究自然杀伤(NK)细胞在单倍体相合骨髓移植中对移植物抗宿主病(GVHD)及造血重建的影响. 方法: 以近交系Balb/c(H-2d)×C57BL/6(H-2b)杂交F1代小鼠(H-2d/b)为供鼠、Balb/c(H-2d)为受鼠,在移植物中增加供者的外周T细胞和(或)NK细胞进行半相合骨髓移植,根据临床表现、病理检查及外周血白细胞(WBC)数,比较不同移植组GVHD发生及造血重建情况. 结果: 骨髓 T细胞组中90%的小鼠出现GVHD,外周血WBC平均在移植后4 wk恢复正常;而骨髓 T 低浓度NK细胞组中20%小鼠出现GVHD, 外周血WBC平均在移植后3 wk恢复正常;骨髓 T 高浓度NK细胞组中仅10%出现GVHD,外周血WBC也平均在移植后3 wk恢复正常. 后两组较骨髓 T细胞组GVHD发生率显著下降(P＜0.01),外周血WBC恢复时间显著提前(P＜0.01). 结论:在小鼠单倍体相合骨髓移植中,同种反应性NK细胞可降低GVHD发生率,促进造血重建.     

单倍体相合骨髓移植白血病患者造血重建的临床分析和实践

目的：观察单倍体相合骨髓移植治疗白血病的造血重建效果。方法选择36例白血病患者作为研究对象,均采取单倍体相合骨髓移植方案进行治疗,对造血重建指标进行观察评估。结果所有患者均在接受骨髓移植后的3~7 d 内白细胞下降至0,中性粒细胞〉0.5×109/ L 和血小板〉20.0×109/ L 的时间分别为（17.5±0.9） d 和（21.5±0.7）d。所有患者3系造血均得到重建。结论骨髓供者通过人粒细胞集落刺激因子预处理后进行骨髓采集,联合多种免疫制剂,能够有效抑制 HLA 多样性屏障,提高白血病患者的造血重建效果,值得重视。     

Graft-versus-leukemia effects from donor lymphocyte infusion after nonmyeloablative allogeneic bone marrow transplantation in mice

Background Nonmyeloablative allogeneic bone marrow transplantation has been used since the 1990s as a new hematological stem cell transplantation strategy for treating hematological diseases. The purpose of this study was to explore the graft-versus-leukemia (GVL) effects of donor lymphocyte infusions (DLIs) after nonmyeloablative allogeneic bone marrow transplantations, while assessing the declines in treatment-associated morbidity, mortality, and graft-versus-host disease (GVHD).Methods A total of 615 (H-2k) mice were injected with L615 tumor cells and received 500 cGy (60Coγ-ray) irradiation three days later, followed by an allogeneic bone marrow transplantation (allo-BMT). The allo-grafts consisted of 3×10（7） bone marrow cells and 1×10（7） spleen cells from BALB/C (H-2d) donor mice. Two days after the allo-BMT, the recipient mice were given 200 mg/kg of cyclophosphamide. Subsequently, recipient mice were infused with either donor spleen cells ［2×10（7）］ on day 14 or 21, or donor spleen cells ［5×10（7）］ pretreated with hydrocortisone and cyclosporin A (CsA) in vitro on day 14 post-BMT.Results The median survival time of mice that received DLI on day 21 and pretreated DLI on day 14 post-BMT was longer than that of controls and the day 14 DLI group (P<0.01). No evidence of severe GVHD was observed in the day 21 DLI group nor in the day 14 treated DLI group. Mixed chimerism was confirmed in the day 14 DLI group, the day 14 treated DLI group, and the day 21 DLI group on the thirteenth day post-transplantation; full donor chimerism was observed two weeks after DLI.Conclusion Donor lymphocyte infusion after nonmyeloablative bone marrow transplantation may reduce transplantation-associated morbidity and mortality while strengthening graft-versus-leukemia effects.     

非清髓性预处理联合髓腔内骨髓细胞输注诱导免疫耐受的实验研究

目的本研究通过非清髓性预处理方案联合髓腔内骨髓移植建立异基因小鼠免疫耐受模型,并探讨其诱导耐受的机理.方法受鼠C57BL/6(B6)于第0天接受60Coγ射线全身照射(TBI,550-cGy),4 h内输注雄性BALB/C(H2d)小鼠来源的骨髓细胞,2 d后腹腔注射环磷酰胺(CTX,200 mg·kg-1).通过皮肤移植、混合淋巴细胞反应(mixed lymphocyte reaction, MLR)检测耐受状态,并应用体外过继转移实验、IL 2逆转实验等探讨免疫耐受机制.结果经骨髓移植的B6小鼠对BALB/C小鼠的皮肤移植物平均存活时间超过300d,较空白组明显延长(P＜0.001);MLR结果证明B6小鼠获得供体特异性耐受,该耐受可以被IL 2逆转且可被过继转移;所有受鼠均未出现GVHD表现.结论非清髓预处理联合髓腔内骨髓移植可以有效诱导异基因小鼠的免疫耐受.     

Successful application of nonmyeloablative stem cell transplantation for paroxysmal nocturnal hemoglobinuria

Paroxysmal nocturnal hemoglobinuria （PNH） is a consequence of nonmalignant clonal expansion of one or several hematopoietic stem cells that have acquired a somatic mutation of phosphatedalinositol glycae-class A （PIGA） gene leading to deficient glycosyl phosphatidylinositol （GPI） synthesis. The pathophysiology of PNH is an abnormality in the glycolipid-anchored proteins of the membranes of blood cells. The absence of complement control proteins, including decay accelerating factor （DAF, CD55） and membrane inhibitor of reactive lysis （MIRL, CD59）, results in intravascular lysis of the abnormal cells. Patients with PNH manifest not only clinical and laboratory signs of chronic hemolytic anemia, but also formation of cytopenia and venous thromboses.[第一段]     

骨髓干细胞移植后mdx鼠抗肌萎缩蛋白的表达

目的研究骨髓干细胞移植治疗Duchenne型肌营养不良鼠(mdx鼠)后抗肌萎缩蛋白表达的动态变化。方法取6~8周龄C57BL/6鼠骨髓干细胞,以2×107个/只干细胞尾静脉移植到经7Gyγ射线预处理后的7~9周龄mdx鼠体内,分别于移植后5、8、12、16周应用免疫荧光、逆转录-聚合酶链反应、Western blot检测腓肠肌抗肌萎缩蛋白的表达情况。结果经γ射线放疗预处理的mdx鼠,在尾静脉移植骨髓干细胞5周后可见抗肌萎缩蛋白少量表达,随移植时间延长表达逐渐增多,16周时肌纤维抗肌萎缩蛋白的表达为12%,并可见肌细胞核中移现象较同龄mdx鼠对照组减少,边居增多。结论同种异体骨髓干细胞移植治疗mdx鼠后,骨髓干细胞通过血液循环趋向病变肌组织,并分化为肌细胞表达抗肌萎缩蛋白。随移植时间延长表达增多,提示骨髓干细胞移植可长久持续参与受损骨骼肌的修复与再生。     

Improvement of cardiac function after transplantation of autologous bone marrow mesenchymal stem cells in patients with acute myocardial infarction

Background The infarct size determines the long-term prognosis of patients with acute myocardial infarction (AMI). There is a growing interest in repairing scar area by transplanting bone marrow stem cells. However, effectiveness of intracoronary injection of bone marrow mesenchymal stem cells (BMSCs) in patients with AMI still remains unclear.Methods Sixty-nine patients with AMI after percutaneous coronary intervention (PCI) were randomly divided into intracoronary injection of BMSCs (n=34) and saline (control group, n=35) groups. Serial single positron emission computer tomography (SPECT), cardiac echo and cardiac electromechanical mapping were done at the designed time intervals until six months after transplantation of BMSCs or injection of saline. Results The proportion with functional defect decreased significantly in the BMSCs patients after three months ［(13±5)%］ compared with that pre-transplantation ［(32±11)%］ and the control group ［(28±10)%］ at three month follow-up (P<0.05, respectively). Wall movement velocity over the infracted region increased significantly in the BMSCs group ［(4.2±2.5) cm/s vs (2.2±1.3) cm/s, P<0.05］, but not in the control group ［(2.2±1.5) cm/s vs (2.7±1.7) cm/s, P>0.05］. Left ventricular ejection fraction (LVEF) three months after transplantation in BMSCs group increased significantly compared with that pre-implantation and with that of the control group at three months post-injection ［(67±11)% vs (49±9)% and (53±8)%, P<0.05 respectively］. SPECT scan results showed that perfusion defect was improved significantly in BMSCs group at three-month follow-up compared with that in the control group ［(134±66)cm2 vs (185±87)cm2, P<0.01］. At the same time, left ventricular end-diastolic volume ［(136±31) ml vs (162±27) ml, P<0.05］ and end-systolic volume ［(63±20) ml vs (88±19) ml, P<0.05］ decreased synchronously. The ratio of end-systolic pressure to end-systolic volume ［Psyst/ESV, (2.84±1.30) mmHg/ml vs (1.72±1.23) mmHg/ml, P<0.05］ increased significantly. Cardiac electromechnical mapping demonstrated significant improvement at three months after implantation of BMSCs compared with that pre-injection in both cardiac mechanical capability as left line local shorting ［LLS, (11.29±1.64)% vs (7.32±1.86)%, P<0.05］ and electrical property as left ventricular endocardial unipolar voltage ［UV, (10.38±1.12) mV vs (7.61±1.09) mV, P<0.01］; perfusion defect decreased from (36.2±6.2) % to (20.3±5.31)% (P<0.01). Twenty-four-hour electrocardiographic monitoring demonstrated no arrhythmias occurred at three-months follow-up.Conclusions The transplantation of BMSCs might improve the cardiac function and it is safe and feasible with no deaths or malignant arrhythmias.