异基因造血干细胞移植后激素耐药胃肠道急性移植物抗宿主病的临床分析

目的探讨异基因造血干细胞移植后激素耐药胃肠道急性移植物抗宿主病(aGVHD)的影响因素。方法回顾性分析西安交通大学第一附属医院2012年1月至2019年12月期间行异基因造血干细胞移植,术后发生胃肠道aGVHD 20例患者的临床资料。按照一线糖皮质激素治疗后的反应分为激素敏感组(13例)和激素耐药组(7例)。单因素Logistics回归分析患者性别、年龄、诊断、移植前微小残留病灶、移植类型、供者年龄、供受者关系、供受者ABO血型、输注单个核细胞数、CD34+细胞数、CD3+细胞数、中性粒细胞及血小板植入时间、CMV及EB病毒感染、胃肠道aGVHD发生的时间等与激素耐药胃肠道aGVHD的关系。观察激素耐药患者治疗后的转归,采用Kaplan-Meier生存分析激素耐药及敏感患者的预后差异。结果20例胃肠道aGVHD患者中7例存在激素耐药。胃肠道GVHD发生时间<1个月,激素耐药的风险增加(OR=13.500,95﹪CI=1.197~152.211,P=0.035),患者性别、年龄、诊断、移植前MRD、移植类型、供者年龄、供受者关系、供受者ABO血型、输注的单个核细胞、CD34+细胞、CD3+细胞、中性粒细胞和血小板植入时间、CMV和EB病毒感染均不影响激素耐药(P>0.05)。7例激素耐药胃肠道aGVHD患者均接受二线CD25单克隆抗体治疗,治疗后5例有效,2例无效死亡。与激素敏感组比较,激素耐药组患者1年总生存率(64﹪比52﹪)降低及无进展生存率(28﹪比32﹪)升高,差异无统计学意义(P>0.05)。结论异基因造血干细胞移植后胃肠道aGVHD激素耐药可能与其发生时间相关,发生时间越早,越容易出现激素耐药。     

Ⅰ类细胞因子超家族新成员--IL-21

IL-21是一种新发现的细胞因子,结构上与IL-2、IL-15相似,可调控T细胞、B细胞增殖,影响NK细胞的分化成熟及其胞毒效应,并参与造血系统的发育;其受体IL-21R 与部分细胞因子共用γ链(γc),可能参与性连锁严重联合免疫缺陷(XSCID)的发病.     

全程干预对造血干细胞移植患者营养状况的影响

目的探讨全程干预对异基因造血干细胞移植患者营养状况的影响。方法将128例异基因造血干细胞移植患者随机分成两组,观察组67例患者营养状况采用全程护理干预,对照组61例患者进行常规护理,并对两组患者移植前和移植后4周的体重、血清白蛋白、前蛋白进行测定,观察Ⅱ°以上口腔黏膜炎及腹泻发生情况。结果两组患者术后4周体重、血清白蛋白、前蛋白、Ⅱ°以上口腔黏膜炎、腹泻比较差异均具有统计学意义(均P0.05)。结论异基因造血干细胞移植患者移植前即开展营养评估、制定营养指导计划,移植期间全程跟踪干预,可降低营养不良程度,减少术后并发症的发生,提高生活质量。     

I类细胞因子超家族新成员——IL-21

IL-21是一种新发现的细胞因子,结构上与IL-2、IL-15相似,可调控T细胞、B细胞增殖,影响NK细胞的分化成熟及其胞毒效应,并参与造血系统的发育;其受体IL-21R与部分细胞因子共用γ链(γc),可能参与性连锁严重联合免疫缺陷(XSCID)的发病。     

异甘草酸镁对急性重症胰腺炎患者血清CAM-1、SIL-2R、IL-2水平及肝肾功能的影响

目的:探讨异甘草酸镁对急性重症胰腺炎患者血清胞间粘附分子-1(CAM-1)、可溶性白细胞介素-2受体(SIL-2R)、白细胞介素-2(IL-2)水平及肝肾功能的影响。方法:选择2014年3月～2016年9月于我院接受治疗的86例急性重症胰腺炎患者,按随机数字表法分为对照组与实验组,每组43例。对照组选用常规治疗,实验组在常规治疗基础上加以异甘草酸镁治疗,两组均持续治疗14天。比较两组的临床疗效,治疗前后血清CAM-1、SIL-2R、IL-2、谷草转氨酶、谷丙转氨酶、尿素氮、血肌酐水平的变化及不良反应的发生情况。结果:治疗后,实验组总有效率为90.69%,显著高于对照组(P0.05);两组血清CAM-1、SIL-2R、IL-2、谷草转氨酶、谷丙转氨酶、尿素氮、血肌酐水平均较治疗前显著降低,且实验组以上指标均明显低于对照组(P0.05)。两组不良反应发生情况比较差异无统计学意义(P0.05)。结论:与常规治疗相比,异甘草酸镁可显著提高急性重症胰腺炎的临床效果,降低血清CAM-1、SIL-2R、IL-2水平,保护肝肾功能。     

甲状腺癌患者血清IL-17、IL-35、SIL-2R表达水平及其临床意义

目的:探讨甲状腺癌患者血清白细胞介素-17(IL-17)、白细胞介素-35(IL-35)及可溶性白介素-2受体(SIL-2R)水平及其对甲状腺癌诊断与病情评估的临床价值。方法:选取我院2015年6月~2016年12月收治的甲状腺腺瘤患者38例、甲状腺癌患者49例为研究对象,另选取同期于我院体检中心接受体检的52例健康体检者为对照组。采用酶联免疫吸附法(ELISA)检测和比较其血清IL-17、IL-35、SIL-2R水平,并分析甲状腺癌患者血清IL-17、IL-35、SIL-2R水平与其年龄、病程、病理分期的相关性。结果:甲状腺腺瘤组血清IL-17、IL-35、SIL-2R水平与对照组比较差异均无统计学意义(P0.05)。甲状腺癌组血清IL-35水平显著低于甲状腺腺瘤组和对照组(P0.01),血清IL-17、SIL-2R水平均显著高于甲状腺瘤组和对照组(P0.01)。血清IL-17、SIL-2R水平随甲状腺癌分化程度的降低而升高,血清IL-35水平随甲状腺癌分化程度的降低而降低(P0.01)。血清IL-17、SIL-2R水平随甲状腺癌病理分期的增加而升高,血清IL-35水平随甲状腺癌病理分期的增加而降低(P0.01)。血清IL-17、SIL-2R水平与甲状腺癌病理分期均呈显著正相关(r=0.432、0.439,P均0.05)。血清IL-35水平与甲状腺癌病理分期呈显著负相关(r=-0.602,P0.05)。血清IL-17与IL-35呈显著负相关(r=-0.323,P0.05),IL-17与SIL-2R呈显著正相关(r=0.429,P0.05),IL-35与SIL-2R呈显著负相关(r=-0.415,P0.05)。结论:甲状腺癌患者的血清IL-17、SIL-2R水平均显著上调,IL-35水平显著下调,其对甲状腺癌的早期诊断、病情评估均具有重要参考价值。     

CD4+CD25+调节性T细胞在造血干细胞移植后诱导免疫耐受的作用

异基因造血干细胞移植是治疗良恶性血液病的有效手段,尤其单倍型移植是具有中国特色的移植体系。然而,移植物抗宿主病(graft-versus-host disease, GVHD)的发生是移植后患者死亡的最重要原因。造血干细胞移植的成功实施需要安全有效的诱导免疫耐受,其中T淋巴细胞在GVHD发生中起到最重要的作用。CD4~+CD25~+调节性T细胞(Treg)是近年来发现的一类能够抑制免疫应答的重要调节性免疫细胞,本文综述了Treg在造血干细胞移植后诱导免疫耐受的作用。     

调节性T细胞及间充质干细胞在急性移植物抗宿主病中的研究进展

急性移植物抗宿主病(acute graft versus host disease,aGVHD)的一线治疗方案为糖皮质激素,但药物治疗特异性差,会增加异基因造血干细胞移植(allogeneic hematopoietic stem cell transplantation,allo-HSCT)后感染和复发的风险,而且激素耐药性移植物抗宿主病的治疗效果欠佳,目前尚缺乏标准的二线治疗方案。近年来,随着对具有免疫调节活性细胞的认识,人们发现此类细胞在aGVHD的防治中可能具有独特的作用,因而应用免疫调节性细胞治疗aGVHD引起了广泛关注,其中以调节性T细胞(regulatory T lymphocyte,Treg)和间充质干细胞(mesenchymal stem cells,MSCs)的研究最为广泛。为更好地了解免疫调节性细胞调控aGVHD的机制以及其临床应用的可行性和有效性,该文主要就Treg和MSCs生物学特性以及Treg和MSCs在预防与治疗aGVHD中的作用作一综述。     

异基因造血干细胞移植治疗耳道复发的急性早幼粒细胞白血病一例并文献复习

目的:探讨急性早幼粒细胞白血病(APL)髓外复发的相关因素及治疗。方法:对1例APL缓解后耳道复发患者的临床资料进行回顾性分析,并复习相关文献。结果:患者2015年8月诊断为APL(低危型),经诱导后达完全缓解,随后进行巩固、维持治疗,并多次行腰椎穿刺术及椎管内注射化疗药物预防中枢神经系统白血病。2017年3月发现左外耳道新生物,活检确诊外耳道髓系肉瘤,示髓外复发。随后出现骨髓复发。经诱导巩固化疗后行异基因造血干细胞移植,存活至今。结论:对于髓外复发的急性早幼粒细胞白血病,其预后较差,异基因造血干细胞移植治疗有较好疗效。     

异基因造血干细胞移植后血小板减少发病机制的研究进展

持续性血小板减少(prolonged thrombocytopenia,PT)是异基因造血干细胞移植后常见的并发症,发生率约为5％～37％.移植患者发生PT不仅会增加出血风险,而且与患者预后不良密切相关,既往回顾性研究表明移植前预处理药物、输注的CD34+细胞数量、移植后移植物抗宿主病和感染等是PT发生的常见危险因素....     

Association of the costimulatory molecule gene polymorphisms and active cytomegalovirus infection in hematopoietic stem cell transplant patients

Determinative associations may exist between costimulatory molecule gene polymorphisms with a variety of post hematopoietic stem cell transplantation (HSCT) viral related clinical outcomes especially acute graft versus host disease (aGVHD). Therefore in this study the associations between costimulatory molecule gene polymorphisms including: cytotoxic T-lymphocyte antigen-4 (CTLA4), programmed cell death-1 (PD-1), inducible T cell costimulator (ICOS), and cluster differentiation 28 (CD28) with active cytomegalovirus (CMV) infection were evaluated in HSCT patients. The 72 allogeneic HSCT patients with and without aGVHD were enrolled in this cross sectional study between years: 2004–2011. The single nucleotide polymorphisms in loci of the costimulatory molecules including: CTLA4 gene (?318 C/T, 1722 T/C, 1661 A/G, +49 A/G), PD-1 gene (PD-1.3 A/G, PD-1.9 C/T), ICOS gene (1720 C/T), and CD28 gene (+17 C/T) were analyzed in studied HSCT patients by PCR-RFLP methods. The active CMV infection was evaluated in fresh EDTA-treated blood samples of each allogeneic HSCT patients by CMV antigenemia kit according to manufacturer’s instruction. Active CMV infection was found in 11 of 72 (15.27 %) of allogeneic HSCT patients. The T allele and TT genotype of the CD28 +17 C/T were significantly higher frequency in active CMV infected allogeneic HSCT patients experienced aGVHD. The G allele and GG genotype of the CTLA4 ?1661 A/G were significantly higher frequent in active CMV infected allogeneic HSCT patients experienced low grade of aGVHD. Finally, finding of significant associations between CD28 +17 C/T and CTLA4 ?1661 A/G genotypes with CMV active infection in allogeneic HSCT patients experienced aGVHD emphasize on the importance of the genetic pattern of costimulatory genes in outcomes of active CMV infection in HSCT patients needs completed studies.     

异基因造血干细胞移植治疗儿童白血病87 例临床分析

目的:评估异基因造血干细胞移植(allogeneic hematopoieticstemcell transplantation,Allo-HSCT)治疗儿童急慢性白血病的效果及相关影响因素。方法:回顾性分析我科2006年2月至2012年2月间,采用Allo-HSCT治疗儿童急慢性白血病87例临床资料,按照白血病类型分为BCR/ABL+ALL、BCR/ABL-ALL、AML、CML组,通过单因素分析和多因素分析考察白血病类型、移植前状态、是否服用伊马替尼治疗和干细胞源等因素对Allo-HSCT治疗效果的影响。结果:所有入组白血病患儿接受Allo-HSCT治疗后,整体生存率(overall survival,OS)59.7%。单因素分析结果显示白血病类型(P=0.023)、a GVHD、服用伊马替尼、移植前状态(P=0.025)和干细胞来源(P=0.003)对患者的整体生存率影响具有统计学意义。多因素分析结果显示干细胞源(PBSC、UCB、BM、BM+PBSC、BM/PBSC+UCB,P=0.046)、移植前状态(CR1、CR2、CR3、NR,P=0.048)和移植种类(同胞、非亲缘、单倍体,P=0.023)能够显著影响OS;而疾病类型(ALL、AML、CML,P=0.083)、性别(P=0.968)、年龄(P=0.847)与生存率关系没有统计学意义。结论:白血病类型、移植种类、a GVHD、干细胞源、是否服用伊马替尼和移植前状态是影响异基因造血干细胞移植治疗小儿白血病疗效的关键因素。     

Adoptive Transfer of Treg Cells Combined with Mesenchymal Stem Cells Facilitates Repopulation of Endogenous Treg Cells in a Murine Acute GVHD Model

Therapeutic effects of combined cell therapy with mesenchymal stem cells (MSCs) and regulatory T cells (Treg cells) have recently been studied in acute graft-versus-host-disease (aGVHD) models. However, the underlying, seemingly synergistic mechanism behind combined cell therapy has not been determined. We investigated the origin of Foxp3⁺ Treg cells and interleukin 17 (IL-17⁺) cells in recipients following allogeneic bone marrow transplantation (allo-BMT) to identify the immunological effects of combined cell therapy. Treg cells were generated from eGFP-expressing C57BL/6 mice (Treg^egfp cells) to distinguish the transferred Treg cells; recipients were then examined at different time points after BMT. Systemic infusion of MSCs and Treg cells improved survival and GVHD scores, effectively downregulating pro-inflammatory Th×and Th17 cells. These therapeutic effects of combined cell therapy resulted in an increased Foxp3⁺ Treg cell population. Compared to single cell therapy, adoptively transferred Treg^egfp cells only showed prolonged survival in the combined cell therapy group on day 21 after allogeneic BMT. In addition, Foxp3⁺ Treg cells, generated endogenously from recipients, significantly increased. Significantly higher levels of Treg^egfp cells were also detected in aGVHD target organs in the combined cell therapy group compared to the Treg cells group. Thus, our data indicate that MSCs may induce the long-term survival of transferred Treg cells, particularly in aGVHD target organs, and may increase the repopulation of endogenous Treg cells in recipients after BMT. Together, these results support the potential of combined cell therapy using MSCs and Treg cells for preventing aGVHD.     

Il-17 mobilizes peripheral blood stem cells with short- and long-term repopulating ability in mice

Autologous and allogeneic bone marrow transplantations have evolved as important cancer therapy modalities. For both indications, peripheral blood has been shown to have distinct advantages over bone marrow as the stem cell source. Cytokine combinations for mobilization have enhanced stem cell yield and accelerated engraftment. However, novel mobilizing agents and strategies are needed to further improve clinical outcomes. Within the donor graft, the dynamic equilibrium between T cells and stem cells critically influences engraftment and transplantation results. IL-17 is a cytokine produced almost exclusively from activated T cells. IL-17 was expressed in vivo with adenovirus technology. Here, proof-of-principle studies demonstrate that IL-17 effectively mobilizes hemopoietic precursor cells (CFU-granulocyte-erythrocyte-macrophage-monocyte, CFU-high proliferative potential) and primitive hemopoietic stem cells (Lin(-/low)c-kit(+)Sca1(+)). Moreover, mouse IL-17 adenovirus-mobilized peripheral blood stem cells rescued lethally irradiated mice. Bone marrow was found to be 45-75% of donor origin at 1 year. In secondary recipients, donor-derived bone marrow cells ranged from 45 to 95%. These data show that IL-17 mobilizes stem cells in mice with short- and long-term reconstituting capacity. Additional comparative studies are needed as well as studies in tumor models to refine distinct potential clinical applications for IL-17-mobilized peripheral blood stem cells.     

CD4+ T cells generated de novo from donor hemopoietic stem cells mediate the evolution from acute to chronic graft-versus-host disease

Acute and chronic graft-versus-host disease (GVHD) remain the major complications limiting the efficacy of allogeneic hemopoietic stem cell transplantation. Chronic GVHD can evolve from acute GVHD, or in some cases may overlap with acute GVHD, but how acute GVHD evolves to chronic GVHD is unknown. In this study, in a classical CD8+ T cell-dependent mouse model, we found that pathogenic donor CD4+ T cells developed from engrafted hemopoietic stem cells (HSCs) in C57BL/6SJL(B6/SJL, H-2(b)) mice suffering from acute GVHD after receiving donor CD8+ T cells and HSCs from C3H.SW mice (H-2(b)). These CD4+ T cells were activated, infiltrated into GVHD target tissues, and produced high levels of IFN-gamma. These in vivo-generated CD4+ T cells caused lesions characteristic of chronic GVHD when adoptively transferred into secondary allogeneic recipients and also caused GVHD when administered into autologous C3H.SW recipients. The in vivo generation of pathogenic CD4+ T cells from engrafted donor HSCs was thymopoiesis dependent. Keratinocyte growth factor treatment improved the reconstitution of recipient thymic dendritic cells in CD8+ T cell-repleted allogeneic hemopoietic stem cell transplantation and prevented the development of pathogenic donor CD4+ T cells. These results suggest that de novo-generated donor CD4+ T cells, arising during acute graft-versus-host reactions, are key contributors to the evolution from acute to chronic GVHD. Preventing or limiting thymic damage may directly ameliorate chronic GVHD.     

Foxp3-expressing T regulatory cells and mast cells in acute graft-versus-host disease of the skin

Acute graft-versus-host disease (aGVHD) limits the effectiveness of allogeneic hematopoietic stem cell transplantation. Foxp3 is required for the development and function of CD4+/CD25+ regulatory T cells (T-regs). Foxp3-expressing T-regs are thought to protect against GVHD. Mast cells are thought to be essential in CD4+/CD25+ regulatory T cell-dependent peripheral tolerance. Twenty biopsies of skin with grades I-III aGVHD were stained for Foxp3 and CD117. Inflammation was quantified by a 4 point scale, 0=no inflammation, 1=50%. T-regs and mast cells were quantified by a 4 point scale, 0=no cells per 20x field, 1=10 cells. T-regs were positively correlated with both inflammation and aGVHD grade. Twelve cases with low T-regs had mild inflammation and lower grades of aGVHD and 6 cases with high T-regs had dense inflammatory infiltrate and higher grades of aGVHD. The number of T-regs, mast cells and density of the inflammatory infiltrate were positively correlated only in cases with mild inflammation. In aGVHD of the skin, T-regs increased with the degree of inflammation and GVHD grade. Mast cells were present at the same density whether aGVHD was of lower or higher grade.     

Combinations of anti-LFA-1, everolimus, anti-CD40 ligand, and allogeneic bone marrow induce central transplantation tolerance through hemopoietic chimerism, including protection from chronic heart allograft rejection

Central transplantation tolerance through hemopoietic chimerism initially requires inhibition of allogeneic stem cell or bone marrow (BM) rejection, as previously achieved in murine models by combinations of T cell costimulation blockade. We have evaluated LFA-1 blockade as part of regimens to support mixed hemopoietic chimerism development upon fully allogeneic BALB/c BM transfer to nonirradiated busulfan-treated B6 recipient mice. Combining anti-LFA-1 with anti-CD40 ligand (CD40L) induced high incidences and levels of stable multilineage hemopoietic chimerism comparable to chimerism achieved with anti-CD40L and everolimus (40-O-(2-hydroxyethyl)-rapamycin) under conditions where neither Ab alone was effective. The combination of anti-LFA-1 with everolimus also resulted in high levels of chimerism, albeit with a lower incidence of stability. Inhibition of acute allograft rejection critically depended on chimerism stability, even if maintained at very low levels around 1%, as was the case for some recipients without busulfan conditioning. Chimerism stability correlated with a significant donor BM-dependent loss of host-derived Vbeta11(+) T cells 3 mo after BM transplantation (Tx). Combinations of anti-CD40L with anti-LFA-1 or everolimus also prevented acute rejection of skin allografts transplanted before established chimerism, albeit not independently of allospecific BMTx. All skin and heart allografts transplanted to stable chimeras 3 and 5 mo after BMTx, respectively, were protected from acute rejection. Moreover, this included prevention of heart allograft vascular intimal thickening ("chronic rejection").