Gene expression of cytokines after allogeneic peripheral stem cell transplantation and its relationship with acute graft-versus-host disease

Objective: To investigate the role of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ in pathogenesis of acute graft-versus-host disease (aGVHD) after allogeneic peripheral blood stem cell transplantation (allo-PBSCT). Methods: Forty-two patients undergoing allo-PBSCT were included in this study. Reverse-transcriptase polymerase chain reaction (RT-PCR) was used to analyze gene expression of cytokines IL-2, IL-4, IL-10, IL-12, and IFN-γ. Results: All patients achieved engraftment, 18 patients developed grade ⅠGVHD, 6 patients developed grade Ⅱ-Ⅳ GVHD. The gene expression of IL-2, IL-12, and IFN-γ increased, the gene expression of IL-4 and IL-10 decreased. Conclusion: Cytokines IL-2, IL-12, and IFN-γ lead to a positive regulation of the development in human aGVHD, and IL-4 and IL-10 play negative regulatory roles.

Fulminant gastrointestinal graft-versus-host disease concomitant with cytomegalovirus infection:Case report and literature review

Here,we report a case of fulminant gastrointestinal graft-versus-host disease(GI-GVHD) with cytomegalovirus(CMV) infection in 44-year-old woman.Despite the difficulties associated with the treatment of GIGVHD and GI-CMV disease,the mucosal findings and the clinical course showed marked improvements during long-term clinical observation.The endoscopic findings were remarkable,with diffuse sloughing mucosa in the stomach and highly active inflammation and deep discrete ulcers throughout the colon.Changes in the CMV quantitative polymerase chain reaction results were correlated with the endoscopic mucosal findings and were useful for assessing the efficacy of the treatment.Although a definite diagnosis of GI-GVHD is generally made by endoscopy with biopsy,the gross appearance of this disease can vary depending on the endoscopy.In this paper,we also conduct a literature review of patients with GI-GVHD.

Acute GvHD and the cutaneous ultrastructural changes in mismatched bone marrow transplantation

Six patients treated with human leukocyte antigen (HLA)-mismatched bone marrow transplantation (BMT) suffered from grade I to IV acute graft-versus-host disease (aGvHD) after engrafting. Up to date, 4 patients with grade I to II GvHD have lived for over 2920, 910, 740 and 680 days, respectively. Two other patients died of grade IV hyperacute GvHD. The results seem to indicate that patients in mismatched BMT have a high incidence of aGvHD within a month. The severity of aGvHD is positively correlated with the degree of HLA mismatching. The higher the degree of mismatch of HLA, the earlier and the more severe the aGvHD occurrs. The cutaneous lesion of the patient with GvHD is severe and of ten complicated by mucositis. Lethal hyperacute GvHD must be considered when a patient shows following signs at beginning: (1) The symptoms appear early (within 2weeks) ;(2) peripheral white blood cell count does not recover (<0. 5×109/L) to normal; and (3) high fever persists. In the epidermal ultrastructure of patients, besides acantholysis, autophagic degeneration of keratinocytes,and satellite cell dyskeratosis, there were scattered necrotic keratinocytes, breaking and thickening of basal membrane and presence of a lot of pigment in the intercellular space. These imply that the ultrastructural damages in the skin of patients with aGvHD after mismatched transplantation are more severe than after matched ones.

Arsenic trioxide alleviates acute graft-versus-host disease by modulating macrophage polarization

This study aimed to explore macrophage polarization in acute graft-versus-host disease after hematopoietic stem cell transplantation, and investigated if arsenic trioxide(ATO) could correct this imbalance. In the colon of GVHD mice, we found that the number of F4/80+iNOS+ cells as well as the expression intensity of TNF-α and IL-1β was greater in the GVHD group than in the BM group, whereas the number of F4/80+CD206+ cells and the expression intensity of IL-10 and TGF-β was greater in the BM group than in the GVHD group. We investigated the effect of ATO on GVHD mice, and found that ATO treatment clearly improved the survival of the mice and reduced the severity of GVHD. In addition, ATO reduced the number of F4/80+iNOS+ cells, and increased the number of F4/80+CD206+ cells in the colon of GVHD mice. Furthermore, ATO sharply decreased CD86 and CD80 expression, and increased CD163 and CD206 expression in macrophages induced from aGVHD patients. Therefore,ATO can modulate the M1 and M2 phenotype in GVHD mice or in macrophages from aGVHD patients. Our data suggest that macrophage polarization is involved in the pathogenesis of aGVHD, and ATO treatment modulates macrophage polarization toward an M2 phenotype.

Risk factors for chronic graft-versus-host disease after anti-thymocyte globulin-based haploidentical hematopoietic stem cell transplantation in acute myeloid leukemia

Chronic graft-versus-host disease(cGVHD)is a major complication following unmanipulated haploidentical hematopoietic stem cell transplantation(haplo-HSCT).We aimed to identify the risk factors for cGVHD in patients who underwent anti-thymocyte globulin-based haplo-HSCT for acute myeloid leukemia(n=280).The diagnosis of cGVHD was in accordance with the National Institutes of Health consensus criteria.A total of 169 patients suffered from cGVHD.The patients who had 3 loci mismatched had a higher 8-year incidence of cGVHD(total,66.0%vs.53.7%,P=0.031;moderate to severe,42.4%vs.30.1%,P=0.036)than the patients who had 1 to 2 loci mismatched.The patients who had maternal donors had a higher 8-year incidence of moderate to severe cGVHD(49.2%vs.32.9%,P=0.024)compared with the patients who had other donors.The patients who had grades III to IV acute GVHD(aGVHD)had higher 8-year incidence of cGVHD(total,88.0%vs.50.4%,P<0.001;moderate to severe,68.0%vs.27.0%,P<0.001)compared with the patients without aGVHD.In multivariate analysis,grades III to IV aGVHD was the only independent risk factor for cGVHD.Thus,further interventions should be considered in patients with severe aGVHD to prevent cGVHD.

诱导aGVHD策略的异基因造血干细胞移植治疗难治复发性急性白血病的初步临床报告

背景与目的:异基因造血干细胞移植治疗难治复发性急性白血病,因移植后复发率和移植相关死亡率高而预后仍较差。本研究旨在探讨异基因造血干细胞移植治疗难治复发性急性白血病的过程中,主动诱导急性移植物抗宿主病(acutegraft-versus-hostdisease,aGVHD)对防止复发的作用。方法:30例成人难治复发性急性白血病,其中急性淋巴细胞白血病16例,急性非淋巴细胞白血病10例,混合性急性白血病4例。移植时第一次完全缓解4例,第二次完全缓解9例,部分缓解12例,未缓解5例。分别实施亲缘性同胞外周血干细胞移植24例(全相合者21例和不相合者3例),非亲缘性全相合移植6例(骨髓移植5例和外周血干细胞移植1例)。所有病例均接受清髓性预处理而强化清除残留白血病细胞的作用。亲缘性全相合者单用环孢素A,亲缘性不相合者或非亲缘性全相合者采取环孢素A+甲氨蝶呤+骁悉+小剂量ATG方法预防aGVHD。移植后+3至+60天仍无aGVHD者,采取每周20%～30%比例逐步减少环孢素A维持剂量及预制性供者淋巴细胞输注等方法诱导移植后早期aGVHD的发生。结果:移植后中位随访18.1月,aGVHD发生率80%(24/30),其中Ⅲ～Ⅳ度aGVHD发生率13.3%(4/30)。在可评价的19例患者中慢性移植物抗宿主病(cGVHD)发生率57.9%(11/19),其中广泛性cGVHD发生率15.8%(3/19)。

雷公藤甲素治疗异基因骨髓移植后aGVHD的实验研究

目的:探索雷公藤甲素(TWH)对异基因HSCT中aGVHD的治疗作用及其机制,并研究雷公藤甲素对移植物抗肿瘤(GVT)作用的影响和对移植物植入的影响。方法:(1)以B6为供者,致死性照射的CB6F1为受者,建立基因半相合移植动物模型;以CB6F1为受者,皮下接种S180细胞并照射,建立基因半相合移植荷瘤aGVHD模型;BALB/c小鼠背部皮下注射S180细胞并照射,建立同基因移植荷瘤动物模型。各组动物分别给予不同剂量TWH及CsA。观察TWH对GVHD、GVT的影响。(2)MTT法测定TWH抑制脾淋巴细胞增殖作用和TWH的体外抑瘤作用。(3)ELISA法测定移植后动物血清IL-4、IFN-γ水平。(4)甲基纤维素半固体培养基检测CFU-GM生成率。结果:(1)小剂量TWH(<1μg/d)降低aGVHD发病率,延长动物生存时间;小剂量TWH与小剂量CsA在预防GVHD方面有协同作用。(2)TWH在抑制aGVHD的同时,保留GVT作用,延长了荷瘤动物生存时间,而无GVHD的同基因移植荷瘤动物生存期缩短。(3)TWH可抑制脾细胞增殖,降低血清IFN-γ水平。(4)一定剂量TWH(<1μg/d)不影响干细胞植入和CFU-GM的产率。结论:TWH具有抑制aGVHD保留GVT的作用,可与CsA协同作用预防异基因HSCT中aGVHD。

异基因造血干细胞移植患者体内环孢素血药浓度与aGVHD的关系

目的研究异基因造血干细胞移植(allo-HSCT)术后急性移植物抗宿主病(aGVHD)的发生与早期环孢素全血浓度及移植结局的相关性。方法回顾性分析了我院2010年1月至2021年6月期间接受allo-HSCT手术的78例患者的环孢素血药浓度,并评估其临床疗效及不良反应。结果术后第1~4周环孢素中位血药浓度分别为332.7(179.8~602.4)ng·mL^(-1),365.5(95.1~568.4)ng·mL^(-1),366.1(65.9~727.9)ng·mL^(-1),316.8(73.8~727.3)ng·mL^(-1);10名患者在中位时间32天时发生了2~4级aGVHD。在单因素分析中,术后第2、3周浓度较高及HLA配型相合的患者发生2~4级aGVHD的概率显著降低;多因素分析显示,术后第3周环孢素浓度(P=0.042;RR=0.993;95%CI,0.986~1.000)及与干细胞供者HLA配型相合与否(P=0.022;RR=14.19;95%CI,2.21~91.05)是影响2~4级aGVHD的发生率的独立因素;2~4级aGVHD的发生率与移植相关死亡率显著性相关(P=0.022)。结论患者在allo-HSCT术后第2~3周保持较高的环孢素血药浓度能显著降低aGVHD的发生率。而aGVHD的发生与移植术后死亡率相关。

Effect and mechanism of acute graft versus host disease on early diffuse murine lung injury following allogeneic stem cell transplantation

To explore the effect and pathogenssis of acute graft-versus-host disease (aGVHD) on early diffuse lung injury in allogeneic hematopoietic stem cell transplantation (allo-HSCT), we established an aGVHD model of C57BL/6→BALB/c mice. Chest computed tomography (CT) scans, histopathology and the levels of cytokines including tumor necrosis factor α (TNFα) and Interferon (IFNγ) in lungs were dynamically detected in recipient mice after transplantation. The incidence of aGVHD was respectively 0%, 0% and 100% in simple irradiation group (A), syngeneic transplant group(B) and allogeneic transplant group (C). Chest CT scans of recipient mice were normal in 3 groups on days +3 and +7 after transplantation. CT showed that two of ten mice had bilateral lung diffuse infiltrate on day +12 (on the brink of death) in group A and 6 of 10 mice had bilateral lung diffuse infiltrate on day +14 (3 d after aGVHD occurring) in group C, and were normal on days +12 and +14 in group B after transplantation. Histopathology of lungs in the 3 groups was similar, consisting of minor interstitial pneumonitis on day +3. Group A showed edema, hyperplasia of epithelial cells and widened alveolar interval on day +7, and epithelial cell necrosis, lymphocyte infiltration, hemorrhage, protein leakage, and local consolidation on day +12. The histopathology of group B showed slight edema of epithelial cells on +7 day, which were slighter than that on day +3, and virtually normal on day +14. The histopathology in group C was characterized by the significant expansion and congestion of capillaries, and lymphocyte infiltration on day +7, the acute pneumonitis was present involving tissue edema, lymphocyte and macrophage infiltration, protein leakage and perivascular inflammation on day +14. In group A, the levels of TNFα were lower on day +7 than on day +3. In group B, the levels of TNFα attained a peak on day +3, which decreased on days +7 and +14. In group C, the levels of TNFα were highest on day +7 and there was a significant difference between those on days +7 and +14 (P=0.816). In group A, the levels of IFNγ on day +7 were higher than on day +3. In group B, the levels of IFNγ increased progressively, but the comparison of IFNγ levels in different times had no statistical significance (P=0.521, 0.118, 0.340). In group C, the levels of IFNγ attained a peak by day +7 and decreased on day +14. aGVHD is the main cause of early non-infectious lung injury. T lymphocytes and TNFα are possibly implicated in the pathogenesis of acute GVHD-induced lung injury. The decreased levels of IFNγ in lung tissues following transplantation might be associated with pulmonary fibrosis in late non-infectious pulmonary complications.

Loss of Lkb1 impairs Treg function and stability to aggravate graft-versus-host disease after bone marrow transplantation

Accumulating evidence suggests that a reduction in the number of Foxp3^(+) regulatory T cells(Tregs)contributes to the pathogenesis of acute graft-versus-host disease(aGVHD),which is a major adverse complication that can occur after allogeneic hematopoietic stem cell transplantation(allo-HSCT).However,the precise features and mechanism underlying the defects in Tregs remain largely unknown.In this study,we demonstrated that Tregs were more dramatically decreased in bone marrow compared with those in peripheral blood from aGVHD patients and that bone marrow Treg defects were negatively associated with hematopoietic reconstitution.Tregs from aGVHD patients exhibited multiple defects,including the instability of Foxp3 expression,especially in response to IL-12,impaired suppressor function,decreased migratory capacity,and increased apoptosis.Transcriptional profiling revealed the downregulation of Lkb1,a previously identified critical regulator of murine Treg identity and metabolism,and murine Lkb1-regulated genes in Tregs from aGVHD patients.Foxp3 expression in human Tregs could be decreased and increased by the knockdown and overexpression of the Lkb1 gene,respectively.Furthermore,a loss-of-function assay in an aGVHD murine model confirmed that Lkb1 deficiency could impair Tregs and aggravate disease severity.These findings reveal that Lkb1 downregulation contributes to multiple defects in Tregs in human aGVHD and highlight the Lkb1-related pathways that could serve as therapeutic targets that may potentially be manipulated to mitigate aGVHD.

异基因造血干细胞移植术后Th9细胞重建特点的研究

目的探讨异基因造血干细胞移植术后Th9细胞重建特点。方法选取接受造血干细胞移植治疗的患者64例为研究对象,其中全相合组31例,单倍体组33例,20名健康人作为对照组,通过FCM和ELISA法动态检测患者及对照组外周血中Th9细胞和IL-9、TGF-β、IL-4的变化特点。结果全相合组和单倍体组的Th9细胞百分比值分别在移植后90d和150d基本恢复正常(P>0.05),IL-9的动态变化趋势与Th9细胞的变化相一致,全相合组IL-9的浓度在移植后60d和90d时高于单倍体组(P<0.05);全相合组TGF-β和IL-4在移植后90d和60d逐渐恢复至正常水平,单倍体组TGF-β和IL-4在移植后150d和60d恢复至正常水平;移植后发生aGVHD患者Th9细胞及IL-9和TGF-β浓度明显低于未发生aGVHD者,差异均有统计学意义(P<0.05)。移植后IL-9的浓度与TGF-β呈正相关。结论全相合组Th9细胞和IL-9的重建早于单倍体组,aGVHD患者Th9/IL-9重建延迟,IL-9可能有助于诱导机体免疫耐受。

allo-HSCT受者细胞因子基因多态性与急性移植物抗宿主病的关系

目的:探讨在异基因造血干细胞移植(allo-HSCT)中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、白细胞介素10(IL-10)、转化生长因子β1(TGF-β1)、干扰素γ(IFN-γ)等多种疾病相关细胞因子基因多态性与急性移植物抗宿主病(aGVHD)的相互关系。方法:选取2014年1月至2015年12月进行allo-HSCT的受者32例及正常人群36例作为研究对象,采用聚合酶链式反应(PCR)联合基因测序对目的基因特殊SNP位点基因分型进行检测,观察受者术后出现aGVHD的不同情况,分析细胞因子基因多态性对allo-HSCT预后的影响,探讨疾病相关细胞因子特殊SNP点突变与a GVHD发病严重程度的潜在相关性。结果:在全部allo-HSCT受者中,TNF-α-308(G/A)、IL-6-174(G/C)、IL-10-1082(A/G)、TGF-β1+915(G/C)、IFN-γ+874(T/A)等细胞因子的基因多态性分布与重度aGVHD的发生率未见有显著性的差异(P>0.05)。在TGF-β1+869SNP位点上,C/T型allo-HSCT患者中重度a GVHD发病率显著高于C/C、T/T两个基因型患者组(P<0.01)。结论:在allo-HSCT患者中TGF-β1+869(C/T)基因多态性与a GVHD发病的严重程度具有密切联系。C/T型allo-HSCT患者更容易发生重度aGVHD,是诱发严重aGVHD出现的潜在危险因素。因此,在allo-HSCT患者中针对TGF-β1+869(C/T)进行基因多态性检测,制定合理的aGVHD预防方案,可能有助于减少减轻aGVHD的发生。

非亲缘异基因骨髓移植术后重症肠道急性移植物抗宿主病的内镜分析

目的分析非亲缘异基因骨髓移植术后并发中、重度结肠炎内镜、病理特征及治疗情况。方法分析总结12例接受骨髓移植术后出现急性移植物抗宿主病(aGVHD)患者的内镜下表现资料。结果4例患者分别于移植后的1～57d并发不同程度中、重度结肠炎,肠镜和病理活检示结肠黏膜充血水肿或上皮层坏死脱落,肠腔正常结构消失,直、结肠多发性溃疡,见较多淋巴细胞和浆细胞浸润,未见巨细胞病毒(CMV)包涵体和巨细胞,诊断为结肠aGVHD。予以糖皮质激素为主的治疗,其中3例予FK506、抗CD3单克隆抗体治疗。2例出现血CMV抗原血症,给予更昔洛韦/磷甲酸钠治疗。1例治疗无效死亡,其余得到有效控制。结论非亲缘异基因骨髓移植后并发aGVHD所致结肠炎,诊断有赖于肠镜和病理活检并为正确治疗提供依据。

抗CD25单克隆抗体治疗非血缘脐血移植后激素耐药的急性GVHD的疗效分析

目的观察抗CD25单克隆抗体（巴利昔单抗）治疗非血缘脐带血造血干细胞移植（UCBT）后糖皮质激素耐药的急性移植物抗宿主病（aGVHD）的疗效。方法 2010年8月—2013年3月21例恶性血液病患者在安徽省立医院接受UCBT后发生中重度急性GVHD（其中Ⅱ度4例、Ⅲ度6例、Ⅳ度11例）,男11例,女10例,中位年龄12（3-33）岁,其中急性淋巴细胞白血病11例,急性髓细胞白血病10例。全部患者均接受环孢素（3 mg·kg^-1·d^-1,静脉滴注）和霉酚酸脂（30 mg·kg^-1·d^-1）并甲基强的松龙1-2 mg·kg^-1·d^-1无效后,加用巴利昔单抗每次20 mg,每周2次静脉点滴,连用3-4次,观察急性GVHD控制情况。结果 21例患者中完全缓解10例,部分缓解为7例,无效4例,总有效率81%。有效患者使用巴利昔单抗后平均起效的时间为6 d,巴利昔单抗开始使用时间和症状缓解开始时间存在相关性。结论抗CD25单抗对UCBT后发生的激素耐药的急性GVHD具有显著的疗效,无明显毒副作用,且应用时间越早,急性GVHD症状缓解越早。

脐带间充质干细胞治疗儿童移植相关并发症的疗效观察

目的探讨脐带间充质干细胞(umbilical cord mesenchymal stem cells,UCMSCs)治疗异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation,allo-HSCT)后儿童急性移植物抗宿主病(acute graft versus host disease,aGVHD)和出血性膀胱炎(hemorrhagic cystitis,HC)的疗效及安全性。方法 AlloHSCT后发生aGVHD 15例(合并HC 5例),难治性aGVHD的13例(合并HC 5例)对一线免疫抑制剂及CD25单抗治疗无效,联合静脉滴注UCMSCs治疗。UCMSCs的平均使用剂量为3.6(1.3~6.0)×106/kg,平均使用次数为5(2~21)次,中位随访时间47(36~73)个月。然后观察aGVHD和HC的恢复、再发情况及UCMSCs治疗相关不良反应。结果 13例难治性aGVHD联合UCMSCs治疗后8例完全缓解、5例部分缓解;5例HC患儿经水化、碱化治疗后效果欠佳,联合静脉滴注UCMSCs治疗后有效。未观察到UCMSCs治疗相关不良反应。结论 Allo-HSCT后联合UCMSCs治疗儿童aGVHD和HC安全有效。

雷帕霉素诱导耐受性树突状细胞抑制急性移植物抗宿主病的动物实验

目的探讨雷帕霉素（RAPA）诱导过继性输注的耐受性树突状细胞（DCs）对急性移植物抗宿主病（a GVHD）的治疗效果。方法分离15只BALB/C雄性小鼠（MHC为H-2Kd）的骨髓细胞及脾细胞,输注入经8.5Gy辐射处理（TBI照射）的30只雄性C57BL/6J小鼠（MHC为H-2Kb）中建立a GVHD模型（BALB/C→C57BL/6J）;为验证a GVHD造模是否成功,将C57BL/6J小鼠分为MHC不合移植组、MHC相合移植组及单纯TBI照射组（10只/组）,3组小鼠经8.5 Gy TBI照射后分别输入BALB/C来源骨髓细胞及脾细胞、C57BL/6J来源骨髓细胞及脾细胞及同等体积PBS对照,采用流式细胞术检测供鼠植入比例并观察受鼠存活情况。体外培养受鼠骨髓来源DCs,加入RAPA以诱导耐受性DCs（RAPA-t DCs）,加入PBS作为对照DCs（PBS-DCs）。将a GVHD模型鼠随机分为3组（10只/组）,分别是RAPA-t DCs实验组（静脉输注RAPA-t DCs 4!106/只）、PBS-DCs对照组（静脉输注PBS-DCs 4!106/只）及PBS对照组（静脉输注PBS）;采用活体荧光成像观察DCs在a GVHD小鼠体内的迁移、分布情况;同时监测3组小鼠的平均体重波动并统计生存率,评价RAPA-t DCs对小鼠a GVHD的抑制效果。结果流式细胞术检测：与PBS-DCs组小鼠比较,RAPA-t DCs组小鼠DCs表面的CD40、CD80、CD86和MHCII表达丰度明显受到抑制（P〈0.05）;2组小鼠DCs表面趋化因子受体CCR1平均荧光强度（MFI）为14.5±1.4 vs.10.0±2.1,表面趋化因子受体CCR5为12.7±2.3 vs 7.2±1.2（P〈0.05）,表面趋化因子受体CCR7为7.8±1.3 vs.6.2±2.5（P〉0.05）。活体成像分析：RAPA处理不影响DCs在体迁移能力,RAPA-t DCs仍可归巢至淋巴结、脾脏等淋巴组织或器官。PBS-DCs组和RAPA-t DCs组a GVHD小鼠在移植后19 d体重（g）下降-2.4±1.5 vs-1.9±1.2（P〈0.05）,生存期（d）延长10.1±5.5 vs 16.6±6.7（P〈0.05）,但各组小鼠最终均全部死亡,整体生存率无差异。结论 RAPA可通过抑制小鼠DCs表面共刺激分子的表达诱导DCs耐受,过继性RAPA-t DCs仍具备一定的T细胞富集区归巢能力;过继性输注RAPAt DCs可一定程度缓解小鼠a GVHD,延长小鼠生存期,但对其整体存活率无明显改善。

减低剂量预处理单倍体外周血造血干细胞移植术后早期免疫重建的临床研究

目的分析新疆医科大学第一附属医院血液病中心制定的减低剂量预处理高剂量非体外去T细胞单倍体外周血造血干细胞移植术(RIC-haplo-HDPSCT)后早期细胞免疫及体液免疫的动态变化特点。方法以行RIC-haplo-HDPSCT移植术的54例患者和20例健康者(对照组)为研究对象,通过流式细胞法检测患者移植术前、移植术后30 d、60 d、90 d及对照组外周血中CD3、CD4、CD8、CD19、CD56细胞的百分比,ELISA法检测移植后30 d、60 d、90 d患者免疫细胞(IgG、IgA、IgM)的百分比,观察RIC-haplo-HDPSCT术后上述免疫细胞的动态变化规律,并比较发生急性移植物抗宿主病(aGVHD)与未发生aGVHD患者各免疫细胞百分比的变化情况。结果(1)在此移植模式下,CD3^(+)T细胞在术后先下降后升高,至移植术后60 d可达对照组水平,60 d后缓慢降低,与对照组相比无差异;CD4^(+)T细胞在移植术后30 d显著下降,随后逐渐恢复,但在术后90 d内均低于对照组;CD8^(+)T细胞在移植术后先降低后显著上升,在移植术后60 d上升至高于对照组水平,之后缓慢下降,与对照组相比无差异;CD19^(+)B细胞在移植术后缓慢上升,但移植术后早期仍明显低于对照组。CD56^(+)NK细胞移植术后30 d时显著高于对照组,30 d后呈下降趋势,与对照组相比无差异。IgG在移植术后30 d升高,然后逐步下降,与对照组相比,差异无统计学意义。IgA及IgM移植术后早期低于对照组,但差异无统计学意义。(2)发生aGVHD患者的CD8^(+)T细胞数量在移植术后30 d时明显高于未发生aGVHD患者。CD56^(+)NK细胞数量在移植术后30 d明显低于未发生aGVHD患者。结论本中心制定的RIC-haplo-HDPSCT移植模式术后NK细胞恢复最快,其次为CD8^(+)T细胞及CD3^(+)T细胞;各体液免疫细胞在移植术后呈下降趋势,但降低并不明显,CD8^(+)T细胞数量增多及CD56^(+)NK细胞的减少可能与aGVHD的发生有关。

脐带间充质干细胞治疗激素难治性急性移植物抗宿主病的临床观察

目的探讨脐带间充质干细胞(UCMSCs)治疗儿童异基因造血干细胞移植(allo-HSCT)后激素难治性急性移植物抗宿主病(SR-aGVHD)的疗效、安全性和生存情况。方法回顾性分析2014年2月—2018年12月在本院血液科行allo-HSCT术后发生SR-aGVHD的患儿临床资料59例,根据治疗过程中是否接受UCMSCs治疗分为UCMSCs组(n=33)和常规组(n=26)。结果UCMSCs组中UCMSCs静脉输注平均细胞数为1.70(0.43-5.78)×106/kg,平均次数为2(1—5)次,均未见输注不良反应。与常规组相比,UCMSCs组在治疗SR-aGVHD的改善中位时间上更短(12 d vs 18 d,P<0.05);2组之间在治疗SR-aGVHD的治愈时间,皮肤、肝脏、胃肠道疗效的总体反应率,单器官和多器官受累的SR-aGVHD疗效的完全缓解率差异均无统计学意义(P>0.05)。UCMSCs组广泛型cGVHD发生率、巨细胞病毒(CMV)感染率低于常规组(P>0.05),EBV感染率及复发率高于常规组(P>0.05)。结合生存曲线,UCMSCs和常规组2组患儿累及生存率无明显差异。结论UCMSCs治疗儿童移植后SR-aGVHD有一定疗效且安全,改善时间更快,对是否会预防CMV感染、广泛型cGVHD的发生及增加移植后EB病毒感染率、复发率和能否改善移植后长期生存需进一步扩大样本研究。

单份与双份非亲缘脐血移植治疗血液疾病临床效果的比较

目的:分析比较单份与双份非亲缘脐血造血干细胞移植治疗血液疾病的临床效果。方法:回顾性分析2006年至2016年国内8家移植中心185例单份脐血干细胞移植(single-unit umbilical cord blood transplantation,SU-UCBT)与66例双份脐血干细胞移植(double-unit umbilical cord blood transplantation,DU-UCBT)病例临床资料,比较SU-UCBT和DU-UCBT 2种移植方式的植入率、中性粒细胞和血小板植入时间,恶性和非恶性血液病患者接受SU-UCBT和DU-UCBT后急性移植物抗宿主病(acute graft vs host disease,a GVHD)发生情况和受者生存率。结果:SU-UCBT和DU-UCBT受者植入率分别为78.9%和70.7%,其中恶性血液病患者植入率SU-UCBT为89.1%,DU-UCBT为82.5%。非恶性血液病患者SU-UCBT和DU-UCBT植入率分别为64.0%和53.8%,SU-UCBT和DU-UCBT的植入率相似(P>0.05)。SU-UCBT和DU-UCBT受者中性粒细胞植入时间中位数都为18 d,血小板植入时间中位数分别为40 d和27 d,无显著差异(P>0.05)。SU-UCBT和DU-UCBT受者Ⅰ、Ⅱ、Ⅲ和Ⅳ度a GVHD发生率分别为22.6%、22.6%、8.2%、12.3%和21.3%、14.9%、10.6%、10.6%,均无显著差异(均P>0.05)。SU-UCBT和DU-UCBT受者中位生存时间分别为1 460 d(17~2 200 d)和988 d(21~2 200 d),6年累积存活率分别为42.5%和40.4%,无显著差异(P>0.05)。结论:SU-UCBT和DU-UCBT植入成功患者的中性粒细胞和血小板植入时间、a GVHD发生率、存活时间及6年累积存活率均无显著差异,提示DU-UCBT是安全有效的移植选择。但仍需完善SU-UCBT和DU-UCBT评价体系及进一步探究DU-UCBT的植入机制,以便指导科学地选择UCBT方式。

A review from mesenchymal stem-cells and their small extracellular vesicles in tissue engineering

This review aims to offer a vision of the clinical reality of cell therapy today in intensive medicine.For this,it has been carried out a description of the properties,functions,and Mesenchymal Stem Cells(MSCS)sources to subsequently address the evidence in preclinical models and studies clinical trials with whole cells and models attributed to small extracellular vesicles(sEVs),nanoparticles made up of microvesicles secreted by cells with an effect on the extracellular matrix,and their impact as an alternative towards cell-free regenerative medicine.MSCs are cells that enhance the regenerative capacity which can be differentiated typically in different lineages committed as bone,cartilage,and adipose tissue.On the other hand,small extracellular vesicles are structures that participate notoriously and crucially in intercellular communication,which has led to a change in the concept of the functions and the role that these vesicles play in living organisms,in the restoration of damaged tissues and the inflammatory response and immunological.We present the mechanisms that are involved in the applications of MSCS as whole cells and their sEVs in cell therapy and cell-free therapy as an alternative in regenerative medicine.Considering the structural loss that occurs after surgical procedures for cystic and tumoral pathology in periodontitis,as well as the maxillary atrophy that determines the rehabilitation with dental implants,it is imperative to find satisfactory solutions.The opportunity provided by the findings in stem cells is a recent introduction in the field of oral surgery,based on the regenerative potential that these cells possess to restore defects at different levels of the oral cavity.This review aims to discover the real applications that stem cells may have in our treatments shortly.