第三方脐血预防异基因造血干细胞移植后移植物抗宿主病

目的 探讨无关供者和单倍型亲缘供者异基因造血干细胞移植(allo-HSCT)前输注第三方脐血预防移植物抗宿主病(GVHD)的效果.方法 2007年至2011年接受无关供者及单倍型亲缘供者allo-HSCT的受者共41例.脐血预防组(25例),于移植前1d予以HLA配型为4/6～6/6的第三方脐血细胞输注,平均输入有核细胞数为(1.64±0.49)×107/kg,次日再输入供者造血干细胞.其余患者作为对照组(16例).两组均采用抗胸腺细胞球蛋白+环孢素A+甲氨蝶呤+吗替麦考酚酯预防急性GVHD.比较两组间急性GVHD的发生率、严重程度以及移植相关死亡率.结果 脐血预防组和对照组aGVHD累积发生率分别为44.0％(11/25)和68.8％(11/16),两组比较,差异无统计学意义(x2=2.403,P＞0.05);Ⅲ～Ⅳ度aGVHD累积发生率分别为16.0％(4/25)和37.5％(6/16),两组比较,差异无统计学意义(x2=2.445,P＞0.05),100 d治疗相关病死率分别为12.0％(3/25)和12.5％(2/16),两组比较,差异无统计学意义(x2=0.002,P＞0.05).结论 在无关供者及单倍型亲缘供者allo-HSCT前应用第三方脐血细胞,可能有效预防GVHD的发生或减轻GVHD的严重度,但尚需要进一步设计扩大规模的随机对照临床试验验证其有效性.     

检测肿瘤坏死因子d基因型预测急性重症移植物抗宿主病

目的探讨肿瘤坏死因子d(TNF-d)基因型在急性移植物抗宿主病(aGVHD)发病过程中的意义。方法选择1997至2000年间进行同胞间骨髓移植的受者51例,根据受者移植后发生aGVHD的程度分为2组。观察组:为Ⅲ/Ⅳ度aGVHD 20例,对照组:为0/Ⅰ度aGVHD 31例。获取受者及其同胞供者的DNA,通过变性高效液相色谱(DHPLC)法结合DNA测序检测TNF-d基因型。结果观察组供者的TNF-d_3/d_3基因型(6/20)检出频率明显高于对照组(2/31);观察组供者的TNF-d_3基因频率(50%)明显高于对照组(27.4%);两组比较,差异均有统计学意义(P<0.05)。观察组受者的TNF-d_3基因频率(45%)明显高于对照组(17.7%),两组比较,差异均有统计学意义(P<0.01)。结论在同胞间的骨髓移植中,供者的TNF-d_3/d_3基因型以及供、受者的TNF-d_3基因频率与Ⅲ/Ⅳ度aGVHD的发生密切相关,检测TNF-d基因型可以预测受者是否发生aGVHD。     

异基因造血干细胞移植后严重的出血性膀胱炎多因素分析

目的 探讨异基因造血干细胞移植（allo-HSCT）后严重的（≥Ⅱ度）出血性膀胱炎（HC）的危险因素。方法 对1997年4月至2004年12月期间的114例allo-HSCT患者的资料进行回顾性分析。以预处理实施之日为观察起点，至移植后＋180 d随访中止。选择11个临床参数，即：年龄、性别、疾病类型、供者类型、预处理方案、移植时疾病状态、急性移植物抗宿主病（aGVHD）、aGVHD的预防、预处理方案中抗胸腺细胞球蛋白（ATG）的应用、中性粒细胞及血小板植活时间做Cox单因素分析。将在单因素分析中P＜0.1作为有统计学意义的因素进行Cox多因素回归分析。移植后180 d内HC累计发生率的计算应用Kaplan-Meier法。结果 （1）114例患者中有29例发生HC，＋180 d内HC的累计发生率为26％，其中Ⅱ级12例，Ⅲ级11例，Ⅳ级6例。（2）单因素分析表明，以下因素与HC的发生密切相关；男性（RR=2.885，P=0.021）、年龄≤25岁（RR=3.265，P=0.002）、Ⅲ～Ⅳ度aGVHD（RR=4.039，P=0.002）、非血缘供者（RR=4．347，P=0.000）、加强的GVHD预防方案（RR=2.218，P=0.045）、疾病进展期（RR=2.668，P=0.009）。（3）对上述有统计学意义的因素进行Cox多因素分析，只有男性（RR=2.993，95％CI 1.218～7.358；P=0.017）和非血缘供者（RR=4.478，95％CI 2.049～9.786；P=0.000）为HC的独立危险因素。结论 男性受者和非血缘供者的造血干细胞移植后发生HC的危险性显著增加。     

异基因造血干细胞移植后白血病髓外复发的临床分析

目的 探讨异基因造血干细胞移植(allo-HSCT)后髓外复发的发病机理、危险因素、治疗方法及临床转归.方法 回顾分析164例allo-HSCT受者的临床资料,选择受者性别、年龄、原发病、移植前疾病状态、是否有髓外浸润、预处理方案、供者类型、HLA相合程度、术后移植物抗宿主病(GVHD)发生情况等10个临床参数做单因素分析,对P＜0.1的单因素进行多因素分析.髓外复发的治疗方法包括局部放疗、单纯手术切除,全身化疗、供者淋巴细胞输注和二次移植.结果 164例受者均顺利重建造血功能.术后发生白血病髓外复发9例(5.5%),髓外复发的中位时间为7.5个月(2.3～42.6个月);术后发生急性GVHD 94例次(57.3%),慢性GVHD 83例次(50.6%).复发后有4例受者死亡.单因素分析表明,受者性别、移植前疾病进展期、移植前髓外浸润、供者类型及术后发生慢性GVHD等因素与白血病髓外复发显著相关(P＜0.1).经Cox回归多因素分析发现,移植前疾病处于进展期(P＜0.05)、白血病髓外浸润(P＜0.01)及术后发生慢性GVHD(P＜0.01)为alloHSCT后白血病髓外复发的独立危险因素.结论 多种因素参与了髓外复发的发病机理,免疫逃逸可能在其中起主要作用.疾病进展期、移植前伴髓外浸润和慢性GVHD是白血病髓外复发的独立危险因素.白血病髓外复发常伴随骨髓复发,预后较差,因此预防白血病细胞由髓外向髓内扩散对长期存活非常重要.     

异基因造血干细胞移植后共刺激分子LIGHT/HVEM的表达与GVHD的关系

目的 探讨异基因造血干细胞移植(allo-HSCT)后共刺激分子LIGHT和HVEM的表达与移植物抗宿主病(GVHD)发生的关系.方法 26例allo-HSCT患者中,预处理方案采用环磷酰胺+足叶乙甙+全身照射11例,采用氟达拉滨+阿糖胞苷+白消安12例,采用环磷酰胺3例.移植时,输注单个核细胞(5.89±3.36)×10~8/kg,CD34~+细胞(3.29±1.29)×10~6/kg.预防急性GVHD(aGVHD)的方案采用环孢素A+短程甲氨蝶呤+吗替麦考酚酯,HLA半相合及非血缘关系者加用抗胸腺细胞球蛋白.GVHD的诊断与分级采用西雅图标准.在预处理前、移植后15 d、发生aGVHD时及aGVHD经治疗好转后分别采集患者外周血2 ml,采用三色流式细胞术进行检测,同时以15名健康志愿者的血液标本作为正常对照.结果 所有患者均获得造血功能重建,移植物均完全植活.移植后共发生aGVHD9例,发生率为35%,其中Ⅲ～Ⅳ度者3例,8例经治疗后好转,1例治疗无效死亡;发生慢性GVHD(cGVHD)7例,发生率为26.9%.患者预处理前和移植后15 d及正常人外周血T淋巴细胞几乎不表达LIGHT,组成性表达HVEM;发生GVHD时,LIGHT表达显著上调,HVEM表达下调;GVHD好转后,LIGHT和HVEM的表达恢复正常.移植后15 d时,发生aGVHD者LIGHT的表达高于未发生aGVHD者,而HVEM的表达较低(P<0.05).Ⅲ～Ⅳ度aGVHD者LIGHT的表达明显高于Ⅰ～Ⅱ度者(P<0.05).结论 LIGHT/HVEM在allo-HSCT后GVHD的发生和发展过程中起重要作用,通过监测该共刺激分子的表达对aGVHD的发生有一定预示作用.     

供者型CIK细胞输注治疗异基因造血干细胞移植后白血病复发

目的 采用受者体内供者型或供者来源的细胞因子诱导杀伤细胞(CIK细胞)过继性免疫治疗异基因造血干细胞移植(allo-HSCT)后白血病复发的效果.方法 回顾性分析2例allo-HSCT后白血病复发患者输注供者型CIK细胞成功开展过继性免疫治疗的临床资料.例1在移植后986 d出现白血病复发,经化疗取得短暂部分缓解,采集受者外周血完全供者型单个核细胞扩增CIK细胞,先后给予受者5个周期的CIK细胞治疗.例2在亲缘allo-HSCT后158 d出现白血病复发,采集亲缘供者外周血单个核细胞扩增CIK细胞,于移植后204 d和294 d给予2个周期的CIK细胞输注治疗.结果 例1经化疗降低白血病细胞负荷后,经过1个周期的CIK细胞输注治疗,获得完全缓解,之后再次给予4个周期CIK细胞治疗,随访7个月取得持续完全缓解.例2复发后停用免疫抑制剂,并行化疗及输注供者干细胞采集物,于移植后187 d出现急性移植物抗宿主病及髓外浸润情况,移植后204 d输注1个周期的CIK细胞后,症状明显减轻.移植后294 d给予第2个周期的CIK细胞治疗,骨痛缓解,现骨髓细胞学持续完全缓解,髓外病灶消失.结论 受者体内供者型或供者来源的CIK细胞可用于allo-HSCT后白血病复发的治疗,效果良好,耐受性好.     

3例造血干细胞移植术后并发重度肠道移植物抗宿主病患者的护理

对2例慢性髓性白血病和1例急性髓性白血病患者行异因因造血干细胞移植(allo-HSCT).干细胞分别来自血缘HLA相合、非血缘HLA相合及血缘HLA 1个位点不合的供者.3例患者于移植后60、105、116 d并发重度肠道移植物抗宿主病(GVHD),予以免疫抑制剂治疗及相应的护理措施,结果1例因肺部结核杆菌及白色念珠菌感染死亡,2例得到有效控制.提示allo-HSCT后并发重度肠道GVHD进展快、症状重,密切观察病情,加强消化道护理、心理护理、保护性隔离及用药护理对移植患者的预后至关重要.     

造血干细胞移植治疗儿童髓系白血病26例疗效分析

目的 了解造血干细胞移植对儿童急性髓系白血病(AML)和慢性粒细胞白血病(CML)的治疗效果.方法 髓系白血病患儿26例,平均年龄为9.8岁,其中CML 8例,AML18例.CML患儿中,第1次慢性期(CPl)6例,加速期(AP)1例,第2次慢性期(CP2)1例;AML患儿中,第1次缓解(CRl)9例,第2次缓解(CR2)7例,2例未缓解(NCR).26例中,2例接受HLA全相合同胞供者的外周血与骨髓干细胞联合移植;2例接受由HLA半相合母亲供者外周血干细胞分离出的CD34+细胞输注;2例接受脐血移植;其余20例接受外周血造血干细胞移植.每例移植有核细胞(6.8±6.0)×108/kg,CD34+细胞(4.0±5.8)×106/kg,CD3+细胞(2.6±3.8)×108/kg.所有患儿均采用白消安及环磷酰胺进行清髓性预处理.移植后采用环孢素A和甲氨蝶呤联用预防移植物抗宿主病(GVHD),接受无关供者造血干细胞移植者加用抗胸腺细胞球蛋白,6例CML患儿加用霉酚酸酯.若发生Ⅱ度以上GVHD,则给予甲泼尼龙或巴利昔单抗治疗.结果 除2例HLA半相合移植失败外,其余24例均获得造血功能重建.24例植入成功的患儿中,2例未发生急性GVHD(aGVHD),15例(62.5%,15/24)出现Ⅰ～Ⅱ度aGVHD,7例(29.2%,7/24)出现Ⅲ～Ⅳ度aGVHD(重度aGVHD).7例重度aGVHD均为CML患儿.26例平均随访20.5个月,其中原发病复发死亡者4例,治疗相关死亡者5例,尚有17例(65.4%,17/26)患儿无病存活.结论 异基因造血干细胞移植有助于提高髓系白血病患儿的存活率,只要加强预防,无关供者造血干细胞移植产生的GVHD是可以控制的.而对于高危患儿,无关供者造血干细胞移植的效果与亲缘相关供者移植相似.     

HLA单倍体相合与全相合外周血造血干细胞移植治疗恶性血液病的疗效比较

目的 观察和比较亲属间人类白细胞抗原(HLA)单倍体相合与全相合外周血造血干细胞移植(PBSCT)治疗恶性血液病的临床疗效.方法 2004年5月至2009年2月,共111例恶性血液病患者进行了异基因PBSCT(allo-PBSCT),其中单倍体相合移植受者51例(单倍体组),同期全相合移植受者60例(全相合组).两组的预处理方案均为清髓性;两组预防移植物抗宿主病(GVHD)均以经典环孢素A加短程甲氨蝶呤作为基础方案,HLA 1个抗原不合时,加用吗替麦考酚酯,HLA 2～3个抗原不合时,再加用抗胸腺细胞球蛋白(ATG)及抗CD25单克隆抗体.移植物为经粒细胞集落刺激因子动员的、未进行体外去除T淋巴细胞的外周血造血干细胞(PBSC).结果 111例受者均获得完全、持久供者干细胞植入.单倍体组和全相合组受者中性粒细胞≥0.5×10~9/L的中位时间分别为14 d和12 d,血小板≥20×10~9/L的中位时间分别为15 d和13 d.单倍体组有25例受者发生急性GVHD(aGVHD),其中Ⅰ度20例,Ⅱ度5例;有33例发生慢性GVHD(cGVHD),其中局限型30例,广泛型3例;4年累积发病率为70.4%;无白血病存活40例,3年预期总无白血病存活率(LFS)为74.5%,其中标危型77.3%,高危型68.2%.全相合组有14例发生aGVHD,其中Ⅰ度10例,Ⅱ度2例,Ⅲ度2例;有37例发生cGVHD,其中局限型32例,广泛型5例;4年累积发病率为58.1%.无白血病存活46例,3年预期总LFS为72.1%,其中标危型77.6%,高危型52.7%.单倍体组受者移植后aGVHD发生率高于全相合组,差异有统计学意义(P<0.05);但cGVHD、原发病复发率和LFS差异均无统计学意义(P>0.05).结论 应用清髓性预处理联合多种免疫抑制剂进行非体外去T淋巴细胞的、亲属间HLA单倍体相合与全相合PBSCT均为治疗恶性血液病安全有效的方案.     

非亲缘HLA表型不全相合供者造血干细胞移植治疗重型β地中海贫血的临床研究

目的探讨非亲缘不全相合供者造血干细胞移植治疗重型β地中海贫血的疗效及安全性。方法回顾性分析2018年1月至2022年4月厦门大学附属中山医院收治的因重型β地中海贫血接受非亲缘不全相合异基因造血干细胞移植(allo-HSCT)15例儿童受者的临床资料。15例均有1～2个HLA位点不相合, 其中男8例, 女7例;年龄7岁(3～12)岁;铁蛋白水平3 417.3(223～14 485)μg/L。预处理采用, 由氟达拉滨(Flu)、白消安(Bu)和环磷酰胺(CTX)组成的FBC方案。移植物抗宿主病(GVHD)防治方案由环孢素(CsA)、吗替麦考酚酯(MMF)、抗人胸腺细胞免疫球蛋白(ATG)联合低剂量后置环磷酰胺(PTCy)和间充质干细胞组成。结果共完成移植15例;截至2022年4月1日, 随访时间为24.1个月(11～49)个月;干细胞植入均成功, 植入分析为完全供者嵌合体。中性粒细胞和血小板植入时间分别是12 d(11～22)d和14 d(8～38)d。死亡2例, 预期2年的总生存率(OS)和无地中海贫血生存率(TFS)为86.67%。其中, 移植后发生Ⅱ度以下急性移植物抗宿主病(aGVH...     

Impact of disparity of minor histocompatibility antigens HA-1, CD31, and CD49b in hematopoietic stem cell transplantation of patients with chronic myeloid leukemia with sibling and unrelated donors

Despite human leukocyte antigen (HLA) identity between donor and recipient, several patients develop acute graft-versus-host disease (aGVHD) after hematopoetic stem cell transplantation (HSCT) because of minor histocompatibility antigen (mHag) incompatibilities. The impact of multiple mHag disparities on the clinical outcome after HSCT still remains to be determined. We studied the genomic polymorphisms of HA-1, CD31, and CD49b and correlated mHag distribution with the occurrence of aGVHD after HSCT from HLA-matched sibling and unrelated donors. All 163 patients examined in our single-center study underwent HSCT for chronic myeloid leukemia in the first chronic phase. HA-1 and CD31 disparities are associated with increased aGVHD incidence in a subgroup of patients who test HLA-B44 supertype positive in univariate analysis. However, in a multivariate analysis, only increased patient age was confirmed as an independent aGVHD risk factor. Our findings indicate that the impact of mHag disparity on aGVHD development in HSCT from HLA-matched sibling and unrelated donors seems to be subordinated to classic aGVHD risk factors.     

RIFLE标准对慢性粒细胞白血病清髓性异基因造血干细胞移植后急性肾损伤的评价及预后分析

目的 探讨慢性粒细胞白血病清髓性异基因造血干细胞移植（HSCT）后急性肾损伤（AKI）的发生率和危险因素及其对患者移植后6个月生存率的影响。 方法 应用RIFLE标准对93例慢性粒细胞白血病患者清髓性异基因HSCT后肾脏功能的变化情况进行回顾性分析。 结果 清髓性异基因HSCT后100 d内有39例（41.9%）患者发生AKI,其中AKI危险（AKI-R）24例（25.8%）,AKI损伤（AKI-I）10例（10.8%）,AKI功能衰竭（AKI-F）5例（5.4%）,中位时间为干细胞回输后40 d（1～96 d）。移植后发生≥Ⅲ度急性移植物抗宿主病（aGVHD）患者与<Ⅲ度aGVHD患者100 d内AKI发生率分别为（81.82±11.63）%和（36.59±5.32）%（P ＝ 0.0037）。移植后出现总胆红素增高患者与无增高患者100 d内AKI发生率分别为（72.73±13.43）%和（37.04±5.37）%（P ＝ 0.0192）。移植后发生≥Ⅲ度aGVHD是患者发生AKI的独立危险因素,其相对危险度（RR）为2.773[95%可信区间（CI）（1.073～7.167）,P ＝ 0.035];并且移植后发生≥Ⅲ度aGVHD患者发生AKI-I和AKI-F的RR为6.320[95%CI（1.464～27.291）,P ＝ 0.013]。移植后发生AKI患者100 d内病死率与无AKI患者差异有统计学意义（P ＝ 0.001）。移植后发生AKI-R、AKI-I和AKI-F的患者6个月的生存率分别为（86.96±7.02）%、（70.0±14.49）%和0（P ＝ 0.000）。 结论 AKI是慢性粒细胞白血病清髓性异基因HSCT后的重要并发症。移植后出现≥Ⅲ度aGVHD和总胆红素增高是发生AKI的影响因素。出现≥Ⅲ度aGVHD的患者易发生较重的AKI。移植后发生AKI程度越严重,患者6个月的生存率越低。RIFLE标准能提高早期诊断AKI的敏感性,并可监测肾功能进展情况,预测预后。     

Are HLA antigens a risk factor for acute GVHD in thalassemic patients receiving HLA-identical stem cell transplantation?

We retrospectively evaluated the association between risk factors and acute graft-versus-host disease (aGVHD) among 182 beta thalassemia patients who received 73 peripheral blood stem cell (PBSC) or 109 bone marrow transplants from HLA-identical siblings between 1991 and 2003. The relationship between the severity of aGVHD was examined for the following factors: HLA antigens, age, sex, ABO mismatch, sex mismatch (between recipient and donor), thalassemia class, graft source, transplant cell dose, CD3+ cell dose, conditioning regimen, GVHD prophylaxis, neutrophil engraftment duration, and blood product transfusions using univariate and multivariate analyses. Overall 61 (34%) patients developed clinical grade III or grade IV aGVHD. Univariate analysis confirmed an increased risk of severe aGVHD, which was associated with HLA-A11, HLA-A26, and PBSCT (P=.04, .03, and .03, respectively). The risk of aGVHD was reduced in the presence of HLA-A3 (P=.03). Multivariate analysis confirmed the increased risk of aGVHD associated with HLA-A11 (P=.04), HLA-A26 (P=.01), and a short-period neutrophil recovery (P=.009). In this study HLA-A11, HLA-A26, PBSCT, and a short neutrophil engraftment period were probable risk factors and HLA-A3 a probable protective factor associated with severe aGVHD. These data may provide useful guidelines to choose strategies for treatment and prevention.     

移植物中不同种类和数量的细胞对异基因外周血造血干细胞移植后急性GVHD的影响

目的 探讨移植物中单个核细胞(MNC),CD34+细胞,T淋巴细胞(包括CD3+、CD3+CD4+、CD3+CD8+和CD4+CD25+),CD3 CD16+CD56+自然杀伤(NK)细胞,以及树突状细胞(DC)Ⅰ和Ⅱ型(DC1和DC2)的数量对人类白细胞抗原(HLA)相合的同胞异基因外周血造血干细胞移植(allo-PBSCT)后急性移植物抗宿主病(aGVHD)的影响.方法 选择65例接受HLA相合的同胞allo-PBSCT的患者进入研究.采用流式细胞术检测移植物中MNC,CD34+细胞,T淋巴细胞(CD3+、CD3+CD4+及CD3+CD8+)的数量,对其中31例患者进一步检测CD4+CD25+T淋巴细胞、CD3-CD16'C+D56+NK细胞及DC的数量.按患者的每公斤体重计算出输注的移植物中以上各细胞的数量.并根据上述细胞数量的中位数分别将患者分为高数量组(>中位数)和低数量组(≤中位数),比较各高数量和低数量组aGVHD的发生情况.结果 CD3+CD4+、CD3+CD8+T淋巴细胞高数量组和相应低数量组相比,Ⅱ～Ⅳ度aGVHD的累积发生率增加,但差异无统计学意义(P值分别为0.089和0.098);CD4+CD25+T淋巴细胞高数量组Ⅲ～Ⅳ度aGVHD的累积发生率显著低于相应低数量组(P<0.05);DC1高数量组总的aGVHD累积发生率显著高于相应低数量组(P<0.05),Ⅱ～Ⅳ度aGVHD累积发生率亦明显高于相应低数量组,但差异无统计学意义(P=0.069).MNC、CD34+细胞、CD3+T淋巴细胞、CD3-CD16+CD56+NK细胞及DC2高数量组与相应低数量组比较,总的及Ⅱ～Ⅳ度aGVHD的累积发生率差异均无统计学意义(P>0.05).结论 移植物中高数量的DCl增加总的aGVHD的累积发生率;而高数量的CD4+CD25+T淋巴细胞则减少Ⅲ～Ⅳ度aGVHD的累积发生率.     

Hla-DPB1 mismatch in HLA-A-B-DRB1 identical sibling donor stem cell transplantation and acute graft-versus-host disease

BACKGROUND: The role of human leukocyte antigen (HLA)-DPB1 as a transplantation antigen is controversial. A higher incidence of acute graft-versus-host disease (aGVHD) has been described after unrelated donor bone marrow transplant when both HLA-DPB1 alleles were mismatched. METHODS: We investigated the impact of a single HLA-DPB1 mismatch after HLA-A-B-DRB1 identical sibling donor transplantation on aGVHD. We analyzed 627 adult patient-donor pairs and identified 30 pairs without HLA-DPB1 identity (4.78%). In 17 cases, the patient had an allele that was not shared by the donor. RESULTS: The cumulative incidence of grades II-IV aGVHD was higher in the HLA-DPB1 mismatched group (66.7% vs. 35.7%, p=0.012). The HLA-DPB1 mismatch was identified by multivariate analysis as an independent risk factor for aGVHD (p=0.020, RR=2.68, 95% CI: 1.73-3.62). CONCLUSIONS: HLA-DPB1 can mediate alloreactive responses. A single HLA-DPB1 mismatch increases the risk of aGVHD after sibling donor stem cell transplantation.     

Gut microbiota intervention by pre and probiotics can induce regulatory T cells and reduce the risk of severe acute GVHD following allogeneic hematopoietic stem cell transplantation

BackgroundAcute graft-versus-host disease (aGVHD) is one of the leading causes of limitation and mortality after allogeneic hematopoietic stem cell transplantation (allo-HSCT). Numerous studies have shown that changes in the gut microbiome diversity increased post-transplant problems, including the occurrence of aGVHD. Probiotics and prebiotics can reconstitute the gut microbiota and thus increase bacterial metabolites such as short-chain fatty acids (SCFAs) that have immunomodulatory effects preventing aGVHD in recipients of allo-HSCTs.Methods/Study DesignWe conducted a pilot randomized clinical trial to investigate whether oral synbiotics are associated with the prevention or reduction in occurrence/severity and mitigate complications of aGVHD following allo-HSCT. A commercially available synbiotic mixture containing high levels of 7 safe bacterial strains plus fructo-oligosaccharides as a prebiotic was administered to allo-HSCT recipients. Out of 40 allo-HSCT patients, 20 received daily a synbiotic 21 days prior to transplantation (days −21 to day 0). In contrast, in the control group 20 recipients of allo-HSCT did not receive a symbiotic therapy.ResultsWithin first 100 days of observation, the incidence of severe (grade III/IV) aGVHD in the a synbiotic-therapy group was 0% (0 out of 20 patients), whereas it was 25% (5 out of 20 patients) in the control group (P = 0.047). The median percentage of CD4 + CD25 + Foxp3+ regulatory T cells (Tregs) among CD4+ lymphocytes on day 28 after HSCT in the synbiotic group was higher (2.54%) than in control group (1.73%; P = 0.01). There was no difference in Treg cells on day 7 after HSCT between two groups. However, the median percentage and the absolute count of Tregs in patients who experience aGVHD was significantly lower on days 7 and 28 after HSCT (both P < 0.05). The overall 12-month survival (OS) rate was higher (90%) in the symbiotic-treated patients than in the control group (75%), but the difference was not statistically significant (P = 0.234).ConclusionOur preliminary findings suggest that synbiotic intake before and during the conditioning regimen of allo-HSCT patients may lead to a reduction in the incidence and severity of aGVHD through the induction of CD4 + CD25 + Foxp3+ regulatory T cells, thus contributing to the improvement of transplant outcomes. Much larger studies are needed to confirm our observations.     

腹腔镜脾切除治疗门静脉高压症及术后并发症的危险因素分析

目的评价腹腔镜脾切除术治疗门静脉高压症术后并发症的发生情况,并分析影响术后并发症发生的危险因素。方法回顾性分析2003年9月至2012年9月四川大学华西医院收治的65例行腹腔镜脾切除术的肝硬化门静脉高压症患者的临床资料。采用改良的Clavien分级系统分析术后并发症,并分析影响患者术后并发症发生的危险因素。计量资料采用t检验,计数资料采用X2检验和Fisher确切概率法,单因素及多因素分析采用Logistic回归模型分析。结果本组患者中,36例行全腹腔镜脾切除术（2例因术中出血中转开腹手术）,29例行手助式腹腔镜脾切除术。术后16例患者发生20例次并发症,行全腹腔镜脾切除术患者术后并发症发生率为38．2％（13／34）,高于行手助式腹腔镜脾切除术患者的10．3％（3／29）,两者比较,差异有统计学意义（X2=3．90,P〈0．05）。按照改良Clavien分级系统,本组患者Ⅰ、Ⅱ、Ⅲa、Ⅲb、Ⅳa、Ⅳb、Ⅴ级并发症分别为4、2、8、1、1、0、0例。全组患者术后住院时间为（8．4±2．9）d,发生Ⅱ、Ⅲ、Ⅳ级并发症的患者术后住院时间为（10．1±3．3）d,长于I级和无并发症患者的（7．7±2．5）d,两者比较,差异有统计学意义（t=4．30,P〈0．05）。单因素分析结果显示：美国麻醉医师协会（ASA）分级和是否使用手助器与术后并发症的发生有关（X2=21．60,5．10,P〈0．05）。多因素分析结果显示：ASAⅢ级和未使用手助器是术后并发症发生的独立危险因素（OR=23．60,4．60,P〈0．05）。ASAm级患者术后并发症发生率是Ⅱ级患者的17．00倍,未使用手助器患者术后并发症发生率是使用手助器患者的5．00倍。结论腹腔镜脾切除术治疗肝硬化门静脉高压症术后并发症的发生率高,但主要集中在ASAⅢ级以下患者,ASA分级和手助器的使用与术后并发症的发生有关。     

Characterization of CD3+CD4-CD8- (double negative) T cells reconstitution in patients following hematopoietic stem-cell transplantation

Background

CD3+CD4−CD8−double negative (DN) T cells, as a distinct subset of regulatory T cells (Tregs), played a pivotal role in patients following hematopoietic stem-cell transplantation.

Methods

This study examines the behavior of CD3+CD4−CD8− double negative (DN) T cells in 73 patients at days 30, 60, 90 and 180 after allo-HSCT.

Results

There was no significant difference in neutrophil and platelet engraftment between the higher and lower absolute counts of 30 days DN Tregs (p = 0.674, 0.863, respectively). The reconstitution of DN Tregs was significantly slower than that of CD8+, CD4+, and CD3+CD8+CD28− T cells (p < 0.001), but significantly faster than that of CD19+ and CD4+CD25+ T cells (p < 0.001, p = 0.032, respectively). Importantly, in the HLA mismatched group, DN Tregs reconstitution had significant effect on aGVHD (p = 0.027) and there was significant correlation between aGVHD and DN Tregs reconstitution (p = 0.035). DN Tregs reconstitution was significantly faster in the patients who were devoid of aGVHD than that of patients who developed aGVHD. Furthermore, we compared the absolute value of DN Tregs at 30 days, 60 days, 90 days and 180 days after allo-HSCT with grade aGVHD and found an inverse linear relationship in the HLA mismatched group (n = 37, P < 0.001, r = − 0.573).

Conclusions

The successful expansion of DN Tregs at 60 days after allo-HCST may help avoid severe manifestations of aGVHD in the HLA mismatched group, suggesting that DN Tregs have potential protection effect against aGVHD.     

Association between CD34+ and CD3+ T-cells in allogeneic grafts and acute graft-versus-host disease in children undergoing allogeneic hematopoietic stem cell transplantation: A single-center study

BackgroundAcute graft-versus-host disease (aGVHD) is a major complication of allogeneic hematopoietic stem cell transplantation (allo-HSCT). We examined the association between the composition of the cell subsets present in allogeneic grafts (allografts) and the occurrence and severity of aGVHD in pediatric patients.MethodsWe retrospectively analyzed 80 consecutive pediatric patients undergoing allo-HSCT at our center.ResultsBoth univariate and multivariate analyses showed that the number of CD34⁺ and CD3⁺ T-cells in allografts were the two highest risk factors associated with II–IV aGVHD. Using receiver operating characteristic analysis, the cutoff levels of the allo-HSCT cell doses were used to divide the recipients into low-dose and high-dose groups. The 100-day cumulative incidence of II-IV aGVHD in the high-dose CD34⁺ and CD3⁺ T-cells group was significantly higher than that of the low-dose group (CD34⁺: 57% vs. 29%, p = 0.009; CD3⁺: 63% vs. 18%, p < 0.001). No other clinical factors or cell subsets correlated with aGVHD incidence.ConclusionsOur analysis indicates that the CD34⁺ and CD3⁺ T-cell numbers in the allografts could be the risk factors for the development of severe aGVHD (level II–IV). Further studies should aim to optimize the critical number of CD34⁺ and CD3⁺ T-cells to reduce the risk of severe aGVHD occurrence in pediatric patients.