抗胸腺细胞球蛋白（ATG）对再生障碍性贫血患者淋巴细胞亚群的影响

利用ＡＰＡＡＰ桥联免疫酶标技术，对３６例接受ＡＴＧ治疗的再生障碍性贫血（ＡＡ）患者进行了淋巴细胞亚群检测，结果发现，ＡＡ患者外周血Ｔ淋巴细胞亚群ＣＤ＾＋３ＣＤ＾＋４细胞治疗前后无明显变化，但ＣＤ＾＋４／ＣＤ＾＋８比值，ＣＤ＾＋８细胞数目治疗前后却有显著性差异。ＡＡ患者外周血ＨＬＡ－ＤＲ＾＋细胞较正常明显增高，但ＡＴＧ治疗后却有所下降，本研究的结果表明，ＣＤ＾＋８，ＨＬＡＤＲ＾＋细胞增高及ＣＤ＾＋４     

调节性T细胞及其分泌的细胞因子在再生障碍性贫血中的意义

目的:检测再障患者外周血中T细胞亚群比例及Treg细胞相关的细胞因子分泌水平,探讨其在再障发病中作用。方法:依据临床诊断标准,收集初发再障组、再障造血恢复组、正常对照组的外周血标本,流式细胞学方法检测各组CD4+、CD8+、NK、NKT、Treg细胞的比例,免疫磁珠法分离各组外周血中Treg细胞,并给予刺激培养,24、48小时后收集上清液,酶联免疫吸附试验(ELISA)检测Treg细胞分泌的细胞因子IL-10、IL-35、TGF-β的表达水平。结果:初发再障组的CD4+,Treg比例与正常对照组和再障造血恢复组相比显著低下;CD8+T细胞比例在再障患者中明显升高,其结果有统计学差异(P<0.05);NK、NKT的细胞与正常对照组比较无明显变化(P>0.05)。分离培养三组人群Treg细胞,ELISA检测结果显示:初发再障患者的IL-10、IL-35、TGF-β表达水平和正常对照组比较明显降低(P<0.05)。结论:T细胞免疫异常是再障发病机制的重要环节,Treg细胞在再障患者恢复造血过程中可能起着正向调控的作用。     

再生障碍性贫血免疫致病机制的研究进展

造血干细胞缺乏导致造血功能衰竭一直被认为是再生障碍性贫血 (ＡＡ)的主要病理机制。最初应用体外造血细胞集落形成证实ＡＡ患者的骨髓中造血细胞明显减少或缺如 ,ＣＤ34 细胞数和集落形成能力均明显低于正常对照组[1] 。然而 ,应用免疫抑制剂如抗胸腺细胞或淋巴细胞球蛋白 (ＡＴＧ或ＡＬＧ)以及环胞菌素Ａ (ＣｙＡ)治疗ＡＡ ,其疗效为 50 %～80 % [2 ,3] ,由此表明既使重型再生障碍性贫血(ＳＡＡ) ,其体内仍有少量造血干细胞的残留 ,且造血干细胞减少可能是免疫功能异常介导的细胞毒性作用所致[4 ,5] 。近年的研究表明 ,由Ｔ淋巴细胞介导…     

再生障碍性贫血患者外周血T细胞亚群和T细胞受体表达水平及其 …

采用流式细胞仪及间接免疫荧光法检测３０例再生障碍性贫血患者外周血中Ｔ细胞亚群及Ｔ细胞表面受体表达水平，并与健康对照组相比，结果表明：７０％再障患者存在ＣＤ４／ＣＤ８比例倒置及ＣＤ８＾＋％异常增高；５０％再障患者外周血γ－δＴ细胞亚群及其在Ｔ淋巴细胞总体中所占比例均显著增高；而αβＴ细胞亚群及ＴｉｒＡ＾＋细胞百分率与正常对照组相比无显著性差异。提示：半数以上再障患者外周血中存在异常增多的γδＴ细胞及     

恶性骨肿瘤患者巨噬细胞功能、NK细胞活性和T细胞亚群的变化

我们检测了３４例恶性骨肿瘤患者的巨噬细胞功能、ＮＫ细胞活性和Ｔ细胞亚群的变化,可为临床上恶性骨肿瘤的免疫治疗及病情判定,提供一定的实验依据。１材料和方法１．１研究对象患恶性骨肿瘤患者３４例,均为经ＥＣＴ检查而确诊的住本院患者,包括：骨肉瘤２０例、滑膜肉瘤１例、软骨肉瘤２例、恶性纤维组织细胞瘤１例、转移癌３例及骨巨细胞瘤７例。其中男２３例,女１１例,年龄１０～８５岁平均３０．０岁。此外,另选择本单位健康人３２例均经查体证实,作为对照组,平均年龄３５．５岁。１．２方法①Ｔ细胞亚群的检测〔１〕：取患者…     

再生障碍性贫血患者骨髓及外周血T、NK细胞膜分子和可溶性分子动态变化

目的研究T淋巴细胞及免疫分子在再生障碍性贫血（AA）致病中的作用及其与临床疗效的相关性。方法应用免疫荧光染色技术和流式细胞仪分析AA骨髓及外周血T淋巴细胞表型,ELISA测定骨髓和外周血白介素-2及其可溶性受体（IL-2,sIL-2R）、可溶性Fas配体（sFas-L）和Flt3配体（FL）的水平。结果（1）AA病人骨髓和外周血CD8+细胞百分率增加,CD4/CD8比例下降;骨髓血CD25+和HLA-DR+细胞百分率增多,急性AA增加尤为显著（P<0.01）;骨髓血中CD16+或CD56+细胞百分率也明显增多（P<0.05）。（2）所有AA患者骨髓及外周血IL-2含量均显著升高,绝大部分患者的sFas-L含量增加,FL水平升高尤为显著,高达正常水平的20倍。IL-2、sFas-L和FL的含量与临床疗效密切相关,经治疗有效的患者骨髓和外周血的IL-2、sFas-L和FL水平明显下降,但FL仍不能降至正常水平。结论T细胞的异常活化,多种免疫分子表达的异常升高,以及产生针对自身造血干/祖细胞的细胞毒效应,是AA造血功能衰竭的主要原因。     

抗人胸腺细胞球蛋白（ATG）对再生障碍性贫血患者淋巴细胞亚群的影响

利用ＡＰＡＡＰ桥联免疫酶标技术，对３６例接受ＡＴＧ治疗的再生障碍性贫血（ＡＡ）患者进行了淋巴细胞亚群检测。结果发现ＡＡ患者外周血Ｔ淋巴细胞亚群ＣＤ３~＋、ＣＤ４~＋细胞治疗前后无明显变化，但ＣＤ４~＋／ＣＤ８~＋比值、ＣＤ８~＋细胞数目治疗前后却有显著性差异。ＡＡ患者外周血ＨＬＡ－ＤＲ~＋细胞较正常明显增高，但ＡＴＧ治疗后却有所下降。本研究的结果表明：ＣＤ８~＋、ＨＬＡ－ＤＲ~＋细胞增高及ＣＤ４~＋／ＣＤ８~＋比值降低在ＡＡ患者的发病机制中起着重要作用。     

再生障碍性贫血患者外周血T细胞亚群和T细胞受体表达水平及其意义

采用流式细胞仪及间接免疫荧光法检测30例再生障碍性贫血患者外周血中T细胞亚群及T细胞表面受体表达水平,并与健康对照组相比,结果表明:70％再障患者存在CD4/CD8比例倒置及CD8~+％异常增高;50％再障患者外周血γδ-T细胞亚群及其在T淋巴细胞总体中所占比例均显著增高;而αβT细胞亚群及TirA~+细胞百分率与正常对照组相比无显著性差异。提示:半数以上再障患者外周血中存在异常增多的γδT细胞及Ts细胞亚群。并可通过其直接或间接作用抑制造血,从而导致再障的发生。     

开胸患者围手术期T细胞亚群及NK细胞的动态变化及意义

目的 :探讨开胸患者围手术期间细胞免疫功能的动态变化及其临床意义。方法 :应用流式细胞仪检测 4 5例开胸患者术前、术后 1、3、5及 7天的外周血T细胞亚群 (CD3、CD4、CD8、CD4 CD8)及NK细胞水平 ,并对 4 5例患者随机分组 ,比较纵隔、肺、食管贲门手术组围手术期T细胞亚群及NK细胞的变化。结果 :与术前相比开胸患者术后 1天CD4 CD8和术后 3天CD4降低。其中 ,纵隔肿瘤手术组术后 1天CD4 CD8和术后 3天CD4降低 ;肺部手术组合术后 1天及 7天CD8和术后 5天NK细胞降低 ,术后 5天CD3及术后 7天CD4 CD8升高 ;食管贲门手术组术后 1天CD4 CD8降低 ,术后 7天CD3及CD4升高。具有统计学意义 (P <0 0 5 )。结论 :开胸患者术后细胞免疫功能呈现受抑制状态而降低 ,主要出现在术后第 1～ 3天 ,术后第 5～ 7天开始恢复。因患者疾病种类及手术类型不同 ,细胞免疫功能受抑制持续的时间及恢复开始的时间不尽相同。食管贲门手术患者术后细胞免疫功能受抑严重且恢复较慢 ,肺部手术患者次之 ,纵隔肿瘤手术患者较轻。开胸患者术后 1周内应用抗菌素防治感染及适当免疫增强治疗是必要的。     

Soluble PD-1 is Associated with Aberrant Regulation of T Cells Activation in Aplastic Anemia

Engagement of the membrane program death-1 (PD-1) receptor by its ligands suppresses T cell proliferation and cytokine production. Aberrant over-expression of costimulatory molecules, including PD-1, has been associated with persistent activation of self-reactive T cells in autoimmune diseases. However, the mechanism underlying the dysfunction of PD-1 in the regulation of T-cell activation in such diseases remains unclear. Here, we report the overexpression of CD4⁺ and CD8⁺ T cell PD-1 and elevated serum levels of soluble PD-1 in aplastic anemia (AA) patients. Detailed characterization of soluble PD-1 revealed that it corresponded to an alternative splice variant PD-1Δex3, which lacks the transmembrane domain but has a soluble extracellular domain of the PD-1 molecule. In a further study, PD-1 fusion protein displayed the ability of increasing the proliferation of T cells in vitro, which suggested that soluble PD-1 might serve as an autoimmune antibody to block the function of membrane-bound PD-1 on T cells and lead to aberrant T cell proliferation. Our study revealed a novel pathogenic pathway in which the function of overexpressed PD-1 to restrict over-self-reaction is counteracted by the excessive production of soluble PD-1.     

Allogeneic Hematopoietic Cell Transplantation in Patients Aged 50Years or Older with Severe Aplastic Anemia

We report on 499 patients with severe aplastic anemia aged ≥ 50years who underwent hematopoietic cell transplantation (HCT) from HLA-matched sibling (n?=?275, 55%) or HLA-matched (8/8) unrelated donors (n?=?187, 37%) between 2005 and 2016. The median age at HCT was 57.8 years; 16% of patients were 65 to 77years old. Multivariable analysis confirmed higher mortality risks for patients with performance score less than 90% (hazard ratio [HR], 1.41; 95% confidence interval [CI], 1.03 to 1.92; P?=?.03) and after unrelated donor transplantation (HR, 1.47; 95% CI, 1 to 2.16; P?=?.05). The 3-year probabilities of survival for patients with performance scores of 90 to 100 and less than 90 after HLA-matched sibling transplant were 66% (range, 57% to 75%) and 57% (range, 47% to 76%), respectively. The corresponding probabilities after HLA-matched unrelated donor transplantation were 57% (range, 48% to 67%) and 48% (range, 36% to 59%). Age at transplantation was not associated with survival, but grades II to IV acute graft-versus-host disease (GVHD) risks were higher for patients aged 65years or older (subdistribution HR [sHR], 1.7; 95% confidence interval, 1.07 to 2.72; P?=?.026). Chronic GVHD was lower with the GVHD prophylaxis regimens calcineurin inhibitor (CNI)?+?methotrexate (sHR, .52; 95% CI, .33 to .81; P?=?.004) and CNI alone or with other agents (sHR, .27; 95% CI, .14 to .53; P < .001) compared with CNI?+?mycophenolate. Although donor availability is modifiable only to a limited extent, choice of GVHD prophylaxis and selection of patients with good performance scores are key for improved outcomes.     

不同宫颈癌临床分期患者外周血中T淋巴细胞及NK细胞的表达分析

目的分析宫颈癌患者外周血中T淋巴细胞及其亚群以及NK细胞的表达情况。方法前瞻性分析145例在我院就诊的宫颈癌患者的临床资料,按照宫颈癌FIGO临床分期差别分为:宫颈癌Ⅰ期38例、宫颈癌Ⅱ期42例、宫颈癌Ⅲ期35例、宫颈癌Ⅳ期30例;同期选择体检正常女性30例作为对照组。应用流式细胞仪检测各组患者外周血CD3^+、CD4^+、CD8^+T细胞亚群、调节性T细胞(Treg)以及NK细胞数量,同时计算各亚群的比例以及Treg占CD4^+T细胞的比例。应用多元Logistic回归分析评估各细胞亚群数量及比例与宫颈癌临床分期的相关性。结果与对照组相比,宫颈癌患者CD3^+T细胞、CD4^+T细胞、NK细胞数量以及CD4^+/CD8^+比值均较低,Treg/CD4^+比值较高(均P <0.05),而CD8^+T细胞数量差异无统计学意义(均P>0. 05)。随宫颈癌病理严重程度递增,CD3^+T细胞、CD4^+T细胞、NK细胞数量以及CD4^+/CD8^+比值逐步降低,Treg/CD4^+比值逐步增高(均P <0.05),而CD8^+T细胞数量差异无统计学意义(均P>0.05)。多元Logistic回归分析显示,Treg/CD4^+比值是宫颈癌临床分期的危险因素,CD4^+T细胞、CD4^+/CD8^+比值及NK细胞是其保护性因素(均P<0.05)。结论宫颈癌患者细胞免疫功能均不同程度降低,晚期患者降低最显著。检测T淋巴细胞亚群及NK细胞可用于宫颈癌患者免疫监测,为临床治疗及预后评估提供参考。     

The Frequency of HLA Class I and II Alleles in Pakistani Patients with Aplastic Anemia

Hematological disorders like Aplastic anemia are quite frequent in Pakistan. Human leukocyte antigen (HLA) system, have been implicated in the development of Aplastic anemia in various population-based studies, The aim of this study was to determine the role of the HLA Class I and Class II alleles in genetic susceptibility to Aplastic anemia in Pakistani patients. HLA A*, B* and DRB1* alleles were analyzed, in 61 Pakistani patients (Females n = 22, Males n = 39) and the control group consisted of 200 ethnically matched individuals (Females n = 89, Males n = 111). The allele frequency of DRB1*15 was found significantly higher in patients 0.36 (p = 0.001 with odds ratio = 1.97), as compared to the controls 0.212. Although DRB1*03 percent frequency was significantly higher in controls 0.175 (p = 0.023) with odds ratio = 0.514, as compared to patients 0.106.Therefore DRB1*15 emerged as a susceptible allele and DRB1*03 as a protective allele in Pakistani Aplastic anemia patients and control samples. No significant difference was found in allele frequencies of other HLA class I and HLA class II alleles for both patients and controls. Three haplotypes A*02 B*40 DRB1*15 (p = 0.021), A*31 B*51 DRB1*13 (p = 0.12) and A*33 B*58 DRB1*15 (p = 0.000) showed significant variations in the two groups.     

Bloodstream Infection after Stem Cell Transplantation in Children with Idiopathic Aplastic Anemia

Bloodstream infection (BSI) is the most common infectious complication of hematopoietic stem cell transplantation (HSCT) and can cause substantial morbidity and mortality. Identification of risk factors for BSI might be helpful in efforts to reduce transplantation-related death. This study analyzed the incidence of BSI and risk factors for BSI after HSCT in pediatric patients with aplastic anemia (AA). BSI occurred in 39 of the 351 patients with AA (11.1%). Onset of BSI occurred at a median of 8 days after HSCT (range, 0 to 92 days). The 5-year overall survival rate was lower in patients with BSI than in patients without BSI (63.32% ± 7.90% versus 93.35% ± 1.44%; P < .0001). Univariate analysis identified the following variables as associated with BSI: history of immunosuppressive therapy with antithymocyte globulin (ATG), transplantation from an unrelated donor, frequent blood transfusion before transplantation, major or major plus minor ABO type mismatch, graft-versus-host disease prophylaxis with tacrolimus and without cyclosporine, and long interval from diagnosis to transplantation. Among these factors, long interval from diagnosis to transplantation was the sole statistically significant risk factor for BSI on multivariate analysis. In patients who underwent HSCT from a related donor, age ≥14 years at transplantation was risk factor for BSI. In contrast, history of immunosuppressive therapy with ATG, frequent blood transfusion before HSCT, graft failure, and major or major plus minor ABO type mismatch were risk factors for BSI in patients who underwent HSCT from an unrelated donor. Because the overall 5-year survival rate without BSI was >90%, even in patients who were received a transplant from an unrelated donor, control of BSI is very important for successful HSCT in pediatric patients with AA.     

Hematopoietic Stem Cell Transplantation in Children and Young Adults with Secondary Myelodysplastic Syndrome and Acute Myelogenous Leukemia after Aplastic Anemia

Secondary myelodysplastic syndrome and acute myelogenous leukemia (sMDS/sAML) are the most serious secondary events occurring after immunosuppressive therapy in patients with aplastic anemia. Here we evaluate the outcome of hematopoietic stem cell transplantation (HSCT) in 17 children and young adults with sMDS/sAML after childhood aplastic anemia. The median interval between the diagnosis of aplastic anemia and the development of sMDS/sAML was 2.9 years (range, 1.2 to 13.0 years). At a median age of 13.1 years (range, 4.4 to 26.7 years), patients underwent HSCT with bone marrow (n = 6) or peripheral blood stem cell (n = 11) grafts from HLA-matched sibling donors (n = 2), mismatched family donors (n = 2), or unrelated donors (n = 13). Monosomy 7 was detected in 13 patients. The preparative regimen consisted of busulfan, cyclophosphamide, and melphalan in 11 patients and other agents in 6 patients. All patients achieved neutrophil engraftment. The cumulative incidence of grade II-IV acute graft-versus-host disease (GVHD) was 47%, and that of chronic GVHD was 70%. Relapse occurred in 1 patient. The major cause of death was transplant-related complication (n = 9). Overall survival and event-free survival at 5 years after HSCT were both 41%. In summary, this study indicates that HSCT is a curative therapy for some patients with sMDS/sAML after aplastic anemia. Future efforts should focus on reducing transplantation-related mortality.     

Outcomes of Hematopoietic Cell Transplantation in Adult Patients with Acquired Aplastic Anemia Using Intermediate-Dose Alemtuzumab-Based Conditioning

Graft-versus-host disease (GVHD) has no therapeutic benefit after hematopoietic cell transplantation (HCT) for patients with acquired aplastic anemia (AA), and its prevention is highly desirable. We designed a conditioning regimen using an intermediate dose of alemtuzumab (50 to 60 mg) and describe our institutional experience of 41 patients who underwent HCT for AA. The median age at HCT was 37 years (range, 17 to 59). The conditioning regimen was high-dose cyclophosphamide (n = 9) or fludarabine based (n = 32). Additional GVHD prophylaxis was with cyclosporine. With a median follow-up of 3.6 years, overall survival at 3 years was 85%. Survival in patients <40 years and ≥40 years was 96% and 67%, respectively (P = .04). Graft failure occurred in 4 (10%) patients; 2 primary and 2 secondary. The cumulative incidences of acute (grades 1 to 2) and chronic GVHD were 27% and 15%, respectively. No patients developed grade 3 to 4 acute GVHD or severe chronic GVHD. The following viral complications were frequent: cytomegalovirus reactivation (79%), herpes simplex (18%), varicella zoster (25%), and BK virus hemorrhagic cystitis (8%). The majority of patients had no significant long-term health issues. This intermediate-dose alemtuzumab-based conditioning regimen results in excellent survival with a favorable impact on GVHD and long-term health outcomes, but close monitoring for viral complications is important.