异基因骨髓移植后嵌合状态的研究进展

骨髓移植是许多血液系统疾病和免疫缺陷性疾病的有效治疗措施，移植成功与否与供细胞在受体内的植入状况即嵌合状态的形成有关。由于造血干细胞输入量、供和受T细胞量、预处理方案以及移植时患病情等因素的影响，可以形成不同形式的嵌合状态。如果受的骨髓或外周血中完全是供的细胞，则形成完全的供嵌合状态（CC）；当检测到供和受两种细胞成分时，则为混合嵌合状态（MC）。不同形式的嵌合状态，以及嵌合状态形式的不同演变趋势，其临床意义各不相同。检测嵌合状态的方法有多种，包括微卫星检测、染色体核型分析、血型转换、红细胞抗原、限制性片段长度多态性、可变数目串联重复序列以及现在最常用的PCR法。本从上述几个方面对嵌合状态研究的进展进行了综述。     

STR-PCR分析嵌合体在同种异基因造血干细胞移植中的应用

利用荧光标记的多重PCR扩增短串联重复序列（STR-PCR）结合毛细管电泳检测供者细胞嵌合率（DC）,以探讨该方法的连续检测对异基因造血干细胞移植（allo-HSCT）后转归的预警作用,采集27例清髓性外周血干细胞移植患者移植前、移植后不同时段的外周血或骨髓,DNA样本用Profiler Plus和Cofiler Plus商品化试剂盒扩增后,用ABI310遗传分析仪进行毛细管电泳,确定基因位点及峰面积,根据基因型的差异选择嵌合率计算公式。结果表明：两种试剂盒测得的DC嵌合率一致;在27对中能区别出供受差别的STR位点,Profiler Plus为6.3（4-9）个,Cofiler Plus为4.9（2-6）个。26例患者均在移植后28天出现供者细胞,1例患者未出现供者细胞。14例患者DC100%,均获得持久植入,至今仍无白血病生存;另有9例患者出现不稳定混合嵌合（MC）状态（DC为0%-90.2%）,其中5例为血液学复发。27例病人中有6例死亡。上述5例复发患者均在出现临床症状前发生DC量下降;供者细胞完全嵌合组移植物抗宿主病（GVHD）的发生率高于MC组。结论：动态检测DC可用于移植动力学研究,对移植物早期植入或被排斥、疾病复发以及GVHD的发生均有预警作用,对早期实施临床干预治疗有重要的指导意义。     

Evaluation of chimerism in DNA samples by PCR amplification of D1S80 with detection by capillary electrophoresis.

BACKGROUND: Analysis of the relative amounts of donor and recipient DNA in bone marrow after bone marrow transplantation is frequently used to determine the status of the transplant. We studied the performance of an assay to quantify chimerism based on amplification of the D1S80 variable number tandem repeat marker by PCR with detection of PCR products by capillary electrophoresis (CE). METHODS AND RESULTS: Samples from potential bone marrow donors and recipients were analyzed separately and in mixtures to simulate various degrees of chimerism from 10% to 90% and subjected to PCR/CE analysis. There was excellent agreement between the measured and known relative proportions of DNA components in chimeric samples. The lower limit of sensitivity for detection of chimerism was 1%; between-runs coefficients of variation were <5%. CONCLUSIONS: Amplification of the D1S80 minisatellite by PCR with CE detection is a reliable method for determination of the relative contribution of different DNAs in mixed samples. This method is fast, quantitative, and extremely reproducible.     

异基因骨髓移植混合嵌合体小鼠模型的构建及影响因素分析

目的:构建小鼠异基因骨髓移植(allogeneic bone marrow transplantation,allo-BMT)混合嵌合体(mixed chimerism,MC)模型,分析影响移植后嵌合水平的相关因素并建立数学模型。方法:通过非清髓allo-BMT联合移植后大剂量环磷酰胺构建MC模型,对构建过程中的123只小鼠进行回顾性研究,对影响其嵌合状态的相关因素进行单因素及多因素Logistic回归分析,并通过R语言进行多元线性回归获得以相关影响因素预测嵌合水平的数学模型。结果:该模型在移植后第14天呈现混合嵌合,第15天供者淋巴细胞(donor lymphocyte infusion,DLI)输注可显著促进后期供体细胞的植入,而PBS对照组植入失败。123只小鼠中,MC和非MC分别有47只(38.21%)、76只(61.79%);单因素分析结果显示,小鼠的基线体重(P=0.001,17.84±1.19 vs 18.50±0.94 g)、预处理条件全身照射(total body irradiation,TBI;P=0.048)及是否加用环磷酰胺(P=0.16)会影响小鼠的嵌合状态,而移植的细胞数及检测时间对其无明显影响。多因素回归分析结果显示,在一定条件下,小鼠基线体重是影响后期嵌合水平的独立因素(OR=0.493,95%CI 0.307-0.791,P=0.003)。经R语言多元线性回归得数学模型:chimerism=6.09-12×weight(g)+80.03×TBI(Gy)-4.4×cell-counts(×10⁷)+0.38×CTX(mg/kg),R²=0.5841,P<0.001。结论:本研究建立了移植后混合嵌合体小鼠模型的构建方法;并建立了影响因素的数学模型。     

Flow cytometric monitoring of red blood cell chimerism after bone marrow transplantation

Chimerism after bone marrow transplantation (BMT) was investigated by flow cytometry analysis of red blood cells (RBCs) and of reticulocytes using a series of selected monoclonal or polyclonal antibodies directed against ABO, Rhesus, Kell, Duffy or MNSs antigens. The method allows the routine detection of less than 0.1% of positive cells in artificial mixed field populations. Blood samples from 135 patients undergoing BMT were investigated around days 15, 30, 45, 60, 90, 180, and then every 6 months after transplantation. Characteristic patterns showing expression of donor red blood cell antigens (expansion markers) and concomitant decrease of recipient specific antigens (depletion markers) within days 16-20 were observed for 125 successfully engrafted patients. Distinct patterns were obtained in 10 patients. A delay in engraftment was evidenced in four patients by the absence of chimerism during the first 6 months without any sign of relapse. Re-appearance of recipient RBCs and reticulocytes was observed in five patients; it was consistent with relapse that was later confirmed by clinical, haematological and cytogenetic studies. Finally, a stable and partial chimerism with 20% of RBCs expressing a marker from the recipient was observed in one patient without any sign of relapse. The reported investigation demonstrated that flow cytometry of RBCs and reticulocytes represents a powerful method to efficiently monitor bone marrow transplanted patients on a long-term basis.     

异基因造血干细胞移植植活状态的变性高效液相色谱检测研究的首次报告

异基因造血干细胞移植后动态检测供者嵌合状态对判断移植效果和实施临床早期干预治疗具有重要意义。在临床上已有多种方法用于植活检测，本研究用变性高效液相色谱(DHPLC)技术结合聚合酶链反应扩增短串联重复序列(STR—PCR)检测造血干细胞移植植活状态的特点及可靠性。用非亲缘个体的系列DNA混合体在体外检测此测定方法的可行性和半定量结果的准确性。结果表明，体外模拟混合嵌合体检测实验，显示扩增前供者DNA嵌合百分率与扩增后供受者DHPLC色谱峰峰面积比值呈直线相关，最小检出DNA百分比为5％。用该方法分析51例移植对的结果显示：45例均至少有2个可以区分彼此的有信息住点(另外5例无移植前受者标本，1例为同卵双生双胞胎)；39例与荧光标记多重PCR扩增STR结合毛细管电泳方法进行对照，准确性为100％；29例与性染色体FISH分析，27例与特殊基因标记的分析结果一致；所有病例均检测到完全供者细胞嵌合状态(FDC)；对3例患者观察到临床复发的同时，检测到供者嵌合体的比例明显下降，其中2例为混合嵌合体，1例转变为受者自身型。在此基础上，我们将该方法首次用于对8例人类白细胞抗原(HLA)配型不同一单倍型亲缘供者干细胞及无血缘关系脐带血混合移植患者植活情况的动态检测。结果显示，8例混合移植患者STR—PCR检测结果均为FDC，来源均为亲缘供者。结论：用DHPLC技术结合短串联重复序列具有快速、经济、无污染和自动化高等特点，用于检测异基因造血干细胞移植物植活状态是可靠的。     

嵌合体的动态定量检测在异基因造血干细胞移植中的应用

目的　建立荧光标记的多重PCR扩增短串联重复序列 (STR PCR)结合毛细管电泳 ,定量检测供体细胞 (DC)嵌合率的方法 ,并探讨该方法的连续定量检测对异基因造血干细胞移植 (allo HSCT)后转归的预警作用。方法　采集 31例接受骨髓移植 (BMT)或非清髓外周血干细胞移植 (NST)患者移植前、移植后不同时段的外周血或骨髓。DNA样本用ProfilerPlus商品化试剂盒扩增后 ,用ABI 310遗传分析仪进行毛细管电泳 ,确定基因位点及峰面积 ,根据供受体基因型的差异选择嵌合率计算公式。结果　 31例患者中 15例 (48.4 % )为性别相合移植 ,只能通过STR PCR进行嵌合体的定量分析 ;性别不合移植患者用STR PCR和荧光原位杂交两种方法定量测得的DC嵌合率一致 ;31对供受体中能区别出供受差别的STR位点有 6 .7(2～ 10 )个 ,所有患者均在移植后 7天 ( 7天 )出现供体来源的细胞 ,BMT组 7天、 14天和 1个月DC中位数均明显低于NST组 ,而在移植中后期无显著性差异。 2 1天时BMT和NST患者均达稳定嵌合 ,DC在 92 %以上 ;中位随访 17(3.5～ 2 9.0 )个月 ,2 6例患者DC≥90 % ,均获得持久植入 ,至今均为无白血病生存。另有 5例患者出现不稳定混合嵌合 (MC)状态 (DC为2 7.3%～ 6 2 .7% ) ,其中 4例复发 ,1例出现移植物被排斥。上述 5例患者均     

The effects of chimeric cells following donor bone marrow infusions as detected by PCR-flow assays in kidney transplant recipients.

40 recipients of first cadaver kidney transplants were given perioperative donor vertebral bone marrow infusions (DBMC), compared with 100 controls who did not receive donor bone marrow. The immunosuppressive regimen included OKT3, Tacrolimus, and steroid maintenance therapy, and, in some patients, newly introduced mycophenolate mofetil. This report describes the 24-mo actuarial follow-up and several immunological monitoring studies including sequential measurements of donor bone marrow lineage subset chimerism by the recently reported PCR-flow assay. This is a sensitive in situ PCR detection system for donor versus recipient histocompatibility genes as well as cell surface CD epitope markers using flow cytometry. The results indicate (a) the stabilization of the donor CD3+ and CD34+ cells in recipient peripheral blood at levels below 1% between 6 mo and 1 yr postoperatively, with a 10-fold higher level of donor cell chimerism of these lineages in recipient iliac crest marrow; (b) significantly lower levels of chimerism in peripheral blood up to 6 mo postoperatively in patients who had early acute (reversible) rejection episodes compared with those who did not; (c) a higher degree of chimerism seen in patients who were class II MHC HLA DR identical with their donors; (d) the identification of a high proportion of the donor bone marrow derived CD3 dimly staining subset of T cells (to which regulatory functions have been ascribed) in recipient peripheral blood and especially in recipient bone marrow; and (e) an unexpectedly increased susceptibility to clinically significant infections (primarily viral), and even death in the DBMC-infused group, compared with controls, but no graft losses because of rejection in the DBMC-infused group. Mixed lymphocyte culture assays showed a trend toward a greater number of nonspecifically low reactors in the DBMC group, as well as a greater number of nonspecifically high reactors in the controls (P = 0.058). The autologous mixed lymphocyte reaction also indicated a trend towards nonspecific immune activation in the DBMC group. Finally, anti-cytomegaloviral IgG antibody reactivity was significantly inhibited in the DBMC group 4-6 mo postoperatively (P = < 0.05). In the controls, there were no donor cell lineages detected by PCR-flow in the peripheral blood. These rather unexpected findings, indicating a more depressed cellular and humoral immune capacity in the DBMC cadaver kidney transplant recipients in this relatively early follow-up period, are discussed relevant to chimerism, MHC restriction, and suppressor activity brought about by specialized DBMC subsets, which still need to be defined.     

Cell-free DNA characteristics and chimerism analysis in patients after allogeneic cell transplantation

Cell-free DNA (cfDNA) isolated from plasma or serum has received increasing interest for diagnostic applications in pregnancy, solid tumors and solid organ transplantation. The reported clinical usefulness of cfDNA obtained from plasma or serum in patients undergoing allogeneic cell transplantation (alloHSCT) is scarce.ObjectiveTo analyze the potential clinical utility of cfDNA chimerism analysis after alloHSCT.Design and methodsA total of 196 samples obtained from 110 patients were investigated for their chimeric status both in peripheral blood and plasma using standard PCR for microsatellite amplification. Plasma DNA size distribution was analyzed using capillary electrophoresis.ResultsThe mean cfDNA concentration in the transplanted patients was 469 ng/ml (range: 50–10,700 ng/ml). The size range of almost 80% of the analyzed fragments was between 80 and 200 bp. In 41 out of the 110 patients included in the study a mixture of donor and recipient plasma cfDNA was detected. There was a statistically significant difference in the percentage of plasma mixed chimerism between the patients without transplant related complications and the patients with either GvHD (p < 0.05) or relapse (p < 0.01). In those patients who showed improvement of GvHD also displayed a decrease in the observable percentage of recipient cfDNA during GvHD treatment. In patients without improvement or even with worsening of acute GvHD, stable or increasing levels of recipient cfDNA were detected.ConclusionscfDNA in combination with peripheral blood and bone marrow cell chimerism analysis might improve its utility in the clinic in particular in those patients with clinical complications after alloHSCT.     

异基因造血干细胞移植后嵌合状态与白血病复发关系的研究

目的 探讨异基因造血干细胞移植（allo HSCT）后嵌合状态与白血病复发的关系。方法 回顾性分析2014年1月至2018年6月于我院行清髓性allo HSCT的成人白血病患者73例,采用短串联重复序列 聚合酶链反应（STR PCR）检测移植后+30、+60、+90天受者骨髓的供受者细胞嵌合率,分析嵌合率与复发的关系。结果 移植后+90天复发组嵌合率低于非复发组。移植后+60 d、+90 d低嵌合患者有较高的复发率（P=0.021,0.027）。多因素分析显示移植前疾病状态、+60d嵌合率、+90d嵌合率是白血病患者行allo HSCT后复发的独立危险因素。结论 Allo HSCT后嵌合状态对复发具有预测价值,+60、+90天低嵌合患者有较高的复发率。     

Induction of lymphoid cell chimerism in noninbred, histocompatible rabbits. A new model for studying allotype suppression in the rabbit

Noninbred rabbits, matched with regard to the major histocompatibility complex (RLA-A and RLA-D loci) but mismatched for Ig allotypes, served as donors (adult) and recipients (newborn) of lymphoid cells. Lasting chimerism regularly followed the transfer of 1 x 10(8)-3 x 10(8) spleen, lymph node, or bone marrow cells, as indicated by the continued production of Ig with allotypic determinants of both donor and recipient. Typically, Ig of donor allotype accounted for 25-50% of total allotypic Ig at 4 wk of age and the amount of donor Ig produced remained stable for up to 20 mo. Total allotypic Ig levels remained normal in the chimeric rabbits. "Chimeric drift" or a gradual diminution of donor products over a period of several months, occurred in some individuals. Transfer of lymphoid cells from allotype- suppressed adult donors to newborns of appropriate allotypes did not result in specific suppression of the target allotype in the recipients. Other experiments showed that lymphoid cells from suppressed donors adoptively transferred to histocompatible recipients continued to synthesize Ig of the nonsuppressed type only. The suitability of using an outbred population of histocompatible but allotype-mismatched rabbits for analyzing allotype suppression and other immunoregulatory phenomena is demonstrated by the results presented here.     

The fate of human Langerhans cells in hematopoietic stem cell transplantation

Langerhans cells (LC) and other antigen-presenting cells are believed to be critical in initiating graft versus host responses that influence the outcome of allogeneic hematopoietic stem cell transplantation. However, their fate in humans is poorly understood. We have sought to define the effect of conditioning regimes and graft versus host disease (GVHD) on the survival of recipient LC and reconstitution of donor cells after transplant. Confocal microscopy of epidermal sheets shows that full intensity transplant (FIT) depletes LC more rapidly than reduced intensity transplant (RIT) at day 0, although the nadir is similar in both at 14-21 d. Recovery occurs rapidly within 40 d in the absence of acute GVHD, but is delayed beyond 100 d when GVHD is active. LC chimerism was determined in sex-mismatched transplants using a two-step Giemsa/fluorescence in situ hybridization assay on isolated cells. Acquisition of donor chimerism at 40 d is more rapid after FIT (97%) than RIT (36.5%), irrespective of blood myeloid engraftment. At 100 d, all transplants achieve at least 90% LC donor chimerism and over half achieve 100%. Complete donor chimerism is associated with prior acute cutaneous GVHD, suggesting a role for allogeneic T cells in promoting LC engraftment.     

供者自然杀伤细胞减轻GVHD并提高移植成活率的实验研究

本研究观察激活的供、受者NK细胞能否减轻骨髓移植后GVHD和增加移植成活率。以C57BL／6小鼠作为供者，将接受6．5Gy^60COγ射线全身照射的BALB／c受鼠随机分为4组，对照1组（无任何处理）、对照2组（骨髓移植）、实验1组（骨髓移植同时输注激活的受者NK细胞）和实验2组（骨髓移植同时输注激活的供者NK细胞），观察每组小鼠的活存期、嵌合率、GVHD临床和病理改变。结果表明：对照1组全部活存，实验1组比对照2组活存期短（P〈0．05），实验2组比对照2组活存期延长（P〈0.01）。实验1组GVHD临床及病理评分高于对照2组（P〈0.05），实验2组GVHD临床及病理评分明显低于对照2组（P〈0.01）。实验1组和实验2组嵌合率均较对照2组高（P〈0．05），实验2组的嵌合率比实验1组明显增加（P〈0．01）。结论：激活的供者NK细胞能延长实验小鼠的活存期，降低GVHD严重程度，增加移植成活率；而激活的受者NK细胞却缩短实验小鼠的活存期，加重GVHD严重程度。受者NK细胞也增加移植成活率，但比供者NK细胞作用小。     

一种基于单核苷酸多态性位点的定量检测异基因造血干细胞移植后嵌合率的新方法

目的 建立一种单核苷酸多态性-聚合酶链反应(SNP-PCR)定量检测异基因造血干细胞移植后供受嵌合率的新方法 ,探讨其可行性、准确性及优越性.方法 利用18个SNP位点筛选移植前每一对供、受者,采用实时定量PCR(RQ-PCR)对筛选出的差异位点行嵌合率定量分析,通过倍比稀释、模拟嵌合与微卫星重复序列-PCR(STR-PCR)、性染色体双色荧光原位杂交(XY-FISH)和融合基因的定量检测,比较、验证方法 的准确性及敏感度.结果①利用内参质粒标准品扩增的17次标准曲线,平均斜率为-3.39,平均截距为39.97,相关系数均＞0.995,扩增效率接近理想水平;批内差及批间差分别为0.50%和1.10%,在可控范围;与模拟混合嵌合相关系数在0.99以上;可重复敏感度达0.01%.②40例移植患者中,95%以上可以筛选出供、受者差异SNP位点;SNP-PCR与STR-PCR结果吻合率达96.7%,与XY-FISH结果比较差异无统计学意义(P＞0.05);SNP-PCR与特定白血病融合基因检测结果比较,完全嵌合(CC)标本均未检出肿瘤相关的融合基因,部分嵌合(MC)标本融合基因均为阳性.结论 SNP-PCR检测嵌合率准确、敏感、易行,具有极高的临床应用前景,克服了STR-PCR竞争抑制及扩增平台期偏倚的缺点,可以替代其进行临床常规检测.     

基于免疫磁珠分选的调节性T细胞亚群嵌合性定量分析

为了探讨免疫分选偶联短串重复序列（STR）方法用于调节性T细胞（Treg）嵌合性定量监测的可行性,以淋巴细胞不同比例混合制备14组人工嵌合体血样,以单个核细胞为起点,以免疫磁珠阴性和阳性选择法获取CD4＋CD25＋Treg细胞,分选后的Treg细胞再进行16位点的STR长度多态性分析。结果显示,从分选后的Treg细胞提取的DNA量能满足STR检测要求,当嵌合体中受体比例≥10%时,16个STR位点均能检出受体和供体特异性等位基因,按照STR峰面积计算的受体比例与理论值相符;当受体比例≤1%时,仅有部分位点能检出受体特异性等位基因,实测嵌合率与理论值存在差异。结论：建立了基于免疫分选的Treg细胞嵌合性定量分析方法,对造血嵌合体的检出灵敏度和准确性在1%-10%,可用于造血干细胞移植术后嵌合体监测。     

非清髓造血干细胞移植后致敏供者淋巴细胞输注对嵌合状态及GVHD影响的实验研究

本研究目的是探讨经受者皮肤致敏后的异基因供者淋巴细胞输注(DLI)是否能在促进形成完全供者嵌合(CC)的同时减轻移植物抗宿主病(GVHD)。以C57BL/6小鼠(H-2b,B6)为受者,于第0天接受60Coγ线全身照射(TBI),总剂量为5.5Gy,照射当天移植经粒细胞集落刺激因子(G-CSF)动员后的BALB/c小鼠(H-2d,BA)外周血干细胞(2×107个),移植后第2天腹腔注射环磷酰胺200mg/kg,并分别于移植后第28天输注致敏/未致敏的供者淋巴细胞2×106。结果显示,致敏后DLI的受鼠(黑色)无1例出现GVHD,60天时转变为完全供者嵌合,表型明显呈现为供鼠(白色)特征,CD4+/CD8+T淋巴细胞比值在DLI后早期下降,半月后有所升高,但仍低于正常水平;未致敏的DLI受鼠出现不同程度的GVHD,嵌合率稍有上升,仍表现为混合嵌合体(MC),CD4+/CD8+比值在DLI后早期升高,后期降至正常水平。结论:经受者皮肤致敏后的DLI在诱导CC的同时降低GVHD发病率,CD4+/CD8+比值与GVHD发病率间具有良好的相关性。     

建立人/猪造血嵌合体模型的初步探讨

本研究目的是利用免疫系统尚未成熟的胎猪和新生猪作为人脐血造血干细胞（HSC）的移植受者，建立人／猪造血嵌合体模型，初步探讨在猪体内扩增HSC的可行性。在无菌隔离器中，通过肌肉或脐静脉（剖子宫）给胎猪移植人脐血细胞，或在断脐前通过脐静脉给新生猪移植人脐血细胞，应用流式细胞术检测移植组猪外周血中人CD45^＋细胞。结果表明：移植后，受试猪生长发育与对照猪一致；移植后60天时，用流式细胞术在猪的外周血中检测到一定比例的人源细胞。结论：胎猪或新生猪能耐受一定量的人源抗原的植入，建立人／猪造血嵌合体模型是可行的。     

Transfusion policy: when to stop the use of extremely rare blood for an allogeneic hematopoietic progenitor cell transplant recipient with a history of red cell alloimmunization

BACKGROUND: Decisions for when to select, and when to discontinue, antigen-negative blood in hematopoietic progenitor cell transplantation (HPCT) recipients with red blood cell (RBC) antibodies can be confusing. In HPCT performed for sickle cell anemia patients who require extremely rare antigen-negative blood, the balance of caution and practicality is further complicated. CASE REPORTS: Four sickle cell anemia patients with current or historic RBC antibodies underwent allogeneic HPC transplantation. One required extremely rare (group O D-, hr(B)-) blood. None of the antibodies caused significant hemolysis after transplant. In the case requiring rare blood, antigen-negative blood was requested after donor RBC engraftment because of incomplete donor white blood cell (WBC) chimerism. CONCLUSIONS: RBC antibodies derived from a recipient of allogeneic HPCT rarely cause significant hemolysis, in contrast to the more severe picture sometimes seen with donor-derived antibodies. When donor WBC chimerism is delayed past the time of donor RBC engraftment, there can be concern for the possibility of future recipient-type antibody production. Even 100 percent donor lymphocyte chimerism is no guarantee of total host plasma cell ablation. Immunoglobulin allotyping, when informative, can suggest chimerism for several years. Recipient-type blood, when extremely rare, may not be available for that duration.     

猕猴非清髓单倍相合造血干细胞移植模型的建立

为了研究用非清髓预处理是否能建立猕猴单倍相合造血干细胞移植模型，采用健康、单倍相合的亲代猕猴为供者，子代为受者。受体用氟达拉滨＋环磷酰胺＋全身照射＋兔抗人胸腺细胞球蛋白作非清髓性预处理；用环胞菌素A、霉酚酸酯、鼠抗人CD25单克隆抗体作为移植物抗宿主病（GVHD）预防方案；第0天输注供者动员后的外周血造血干细胞；定期监测造血恢复、造血嵌合水平和GVHD发生等情况。结果表明：4例猕猴用非清髓性预处理后，移植后8天内造血均能恢复，早期均有供者造血干细胞植入；例3、例4植入成功，在移植后12、14天出现Ⅱ-Ⅲ度GVHD；例1在移植后7天低比例供者植入，最后出现移植排斥；例2在移植后7天供者成分占50％，后因肾衰早期死亡。结论：用非清髓性预处理可以跨越单倍相合猕猴的MHC屏障，成功建立了单倍相合造血干细胞移植的模型。为进一步买验研究奠定了基础。