异基因造血干细胞移植后白血病复发伴活动性移植物抗宿主病患者的临床研究

目的 探讨异基因造血干细胞移植(allo-HSCT)后白血病复发伴活动性移植物抗宿主病(GVHD)患者GVHD与GVL效应的分离.方法 分析11例接受allo-HSCT后在白血病复发时存在活动性GVHD的患者其原发病、疾病状态、复发时GVHD类型、供者淋巴细胞输注(DLI)疗效及转归等对GVL效应的影响.结果 11例患者包括急性淋巴细胞白血病5例,急性髓系白血病6例,其中5例曾行预防性DLI,复发时伴有活动性DLI后GVHD,包括2例Ⅱ度急性GVHD(aGVHD),1例原局限型慢性GVHD(cGVHD)加重+新发Ⅱ度aGVHD,2例广泛型cGVHD;这5例患者复发后,2例行化疗+治疗性DLI,DLI后在广泛型cGVHD反复加重情况下,1例达完全缓解(CR)后再次复发,1例未达CR.另6例患者复发前未行预防性DLI,白血病复发时亦均存在活动性GVHD,包括3例广泛型cGVHD、1例Ⅰ度aGVHD及2例Ⅲ～Ⅳ度aGVHD,复发后行化疗+治疗性DLI,之后2例达CR后再次复发,4例未达CR.结论 allo-HSCT后活动性GVHD不一定伴随可抑制白血病复发的有效GVL效应.     

Th17细胞在小鼠移植物抗宿主病发病早期中的作用

目的 探讨Th17细胞在急性移植物抗宿主病(aGVHD)发病早期的作用及机制.方法 以C57/B6为供鼠,BABL/c为受鼠,致死性全身照射(TBI)后输注供鼠骨髓细胞和脾细胞混合细胞悬液,建立小鼠aGVHD模型.实验分为4组:正常对照组;单纯照射组(IRD);异基因骨髓移植(alloBMT)+二甲基亚砜(DMSO)组;allo-BMT+常山酮(HF)组.从-1 d开始到+10 d,观察小鼠弓背、翘毛、体质量变化等aGVHD的表现并进行临床评分.动态观察小鼠aGVHD发生及存活情况.取小鼠的肝、小肠和皮肤进行病理学分析.用流式细胞术检测移植后小鼠Th1/Th17亚群的变化.结果 移植后实验组小鼠均发生aGVHD,allo-BMT+HF组小鼠aGVHD表现较allo-BMT+DMSO组小鼠严重.病理学分析示allo-BMT+HF组小鼠皮肤损伤较轻,而肝脏和小肠损伤较重,与allo-BMT+DMSO组比较,+6 d allo-BMT+HF组Th17细胞比例明显下降(分别为1.04%和0.71%),Th1比例明显升高(分别为5.35%和12.81%),Th1/Th17比值显著升高(P<0.05).结论 异基因造血干细胞移植早期阻断Th17导致Th1比例明显升高(5.31和18.07),加重aGVHD.

Abstract：

Objective To explore the role of Th17 cells in early onset of acute graft-versus-host disease(aGVHD) and its mechanism. Methods Mice aGVHD model was established by irradiated BABL/c mice inoculated with mixed suspension of C57BL/6 bone marrow cells and splenocytes. The mice were divided into 4 groups:①normal control,②irradiated group, ③allo-BMT + DMSO group,④allo-BMT + halofugine (HF) group. HF was given intraperitoneally at 5 μg per mouse from -1 d to + 10 d after allogeneic bone marrow transplantation(allo-BMT). Mice aGVHD symptoms and survival were observed. Th1/Th17 cells ratio was evaluated by flow cytometry. Results All the experimental groups ③ and ④developed aGVHD after transplantation. More severe aGVHD was observed in group ④than in group ③. HF prevented cutaneous aGVHD in all the mice, but augmented hepatic and small intestine GVHD. The percentage of Th17 cells and the ratio of Th1/Th17 were significantly higher while the percentage of Th1 cells was significantly lower in group ④ at day+6(P<0.05). Conclusion Early blockage of Th17 cell results in increase of Th1 cell percentage, which exacerbates aGVHD.     

小剂量甲氨蝶呤联合小剂量糖皮质激素与标准剂量糖皮质激素治疗急性移植物抗宿主病的比较

目的 比较小剂量甲氨蝶呤(MTX)联合小剂量糖皮质激素与标准剂量糖皮质激素治疗急性移植物抗宿主病(aGVHD)的疗效和安全性.方法 78例骨髓移植后发生aGVHD患者,按不同治疗方案分成两个同期对照研究组,32例患者采用A方案即小剂量MTX(每次10 mg)联合小剂量甲泼尼龙(0.5 mg·kg-1·d-1)治疗aGVHD,46例患者采用B方案即单用甲泼尼龙(1～2 mg·kg-1·d-1)治疗aGVHD,治疗后对临床疗效和安全性进行评估.结果 两方案对不同严重程度aGVHD的疗效以及对不同累及部位aCVHD的疗效均相似,差异无统计学意义.两方案相比较,需要二线治疗、发生细菌感染、真菌感染、病毒感染、复发及移植相关死亡率类似,差异无统计学意义.A、B两方案发生慢性移植物抗宿主病(cGVHD)分别为56.25%和34.78%,差异无统计学意义(P=0.068).A方案3例患者血象下降,在停药后血象很快恢复;B方案3例患者出现可逆性血糖升高,激素减停后可摆脱胰岛素治疗,血糖逐渐恢复正常.结论 两种方案治疗的疗效类似,安全性均较好.

Abstract：

Objective To compare the efficacy and safety of a Iow dose (LD) methotrexate (MTX) in combination with a LD methylprednisolone(MP) or standard dose(SD) MP in the treatment of acute graftversus-host disease (aGVHD).Methods Seventy-eight developing aGVHD patients after allogeneic bone marrow transplant(allo-BMT) were divided into two contemporaneous research groups: Group A, 32 patients received intravenous MTX at a dose of 10 mg plus LD-MP (0.5 mg · kg-1 · d-1 ); Group B, 46 patients received SD-MP ( 1 - 2 mg · kg-1 · d-1 ).Results The overall treatment response rate in different degree aGVHD or in different involved aGVHD was similar in two groups, and so was the probability of second-line treatment, fungal infections, bacterial infections, cytomegalovirus(CMV) infection, relapse and transplantrelated mortality (TRM).There was no statistical difference between the two groups.Chronic graft-versushost disease(cGVHD) were 56.25% in Group A versus 34.78% in Group B,being no statistically signifcantly different(P = O.068).In group A three of the 32 patients developed severe leukopenia and/or severe thrombocytopenia and their blood cell counts quickly returned to baseline values after drug withdrawal.In group B three of the 46 patients developed reversible hyperglycemia, after drug withdrawal, they were independent of insulin and blood glucose returned to normal values.Conclusion The efficacy of two groups are silmilar and both safety.     

食管鳞状细胞癌患者血清T辅助细胞类细胞因子的表达及其临床意义

目的 探讨食管癌患者血清T辅助细胞(Th1、Th2)类细胞因子的表达及其临床意义.方法 采用双抗体酶联免疫吸附法(ELISA)检测45例食管鳞状细胞癌患者及30名健康体检者外周血Th1[干扰素(INF)-γ、白细胞介素(IL)-2]、Th2类(IL-4)细胞因子的含量.结果 食管癌患者Th1类细胞因子INF-γ、IL-2表达含量较健康对照组降低[(11.51±2.15)、(16.31±2.27)ng/L,t=18.24,P＜0.01);(10.20±2.10)、(16.92±2.41)ng/L,t=15.31,P＜0.05];Th2类细胞因子(IL-4)较健康对照组升高[(5.30 -0.11)、(2.23±0.29)ng/L,t=5.79,P＜0.05],且其表达趋势与肿瘤的TNM分期有关.结论 食管癌患者体内存在Th1/Th2漂移现象.该现象可能与肿瘤细胞发生免疫逃逸有关.

Abstract：

Objective To study the expression of Th1 and Th2 types cytokines in patients with esophageal squamous cell carcinoma and the clinical significance. Methods The expressions of Th1 ( INF-γ、IL-2) and Th2 (IL-4) types cytokines in cultured peripheral blood mononuclear cells (PBMC) from 30 healthy controls and 45 esophageal squamous cell carcinoma patients were determined by Enzyme-linked immunosorbent assay (ELISA). Results Compared with the control group, the expressions of Th1 type cytokines, IL-2 and INF-γwere significantly lower in the cultured PBMCs from the cancer patients (tINF-γ= 18. 24 ,tIL-2 = 15.31 ,Ps ＜0. 01 ) ,while the expression of Th2 type cytokines,IL-4 was significantly higher in the cultured PBMCs from the cancer patients ( tIL-4 = 5.79, P ＜ 0. 05 ). The expression changes of the cytokines showed a correlation with TNM (tumou-node-metastasis) stage of the tumor. Conclusion The Th1/Th2 shift existed in patients with esophageal squamous cell carcinoma and it might be one of the underline mechanisms of tumor immune escape.     

造血干细胞移植治疗重型再生障碍性贫血

Objective To evaluate the therapeutic effects of allogeneic hematopoietic stem cell transplantat (allo-HSCT) for severe aplastic anemia (SAA). Methods Four patients of SAA underwent allo-HSCT at the bonemarrow transplant unit in our hospital from March 2003 to May 2009. Stem cell source was an HLA (human leukocyte antigen) matched related donor (MRD) in 3, HLA 1 (B) mismatched related donor in 1 patient A retrospective analysis was performed on interval from diagnosis to transplant,HSCT manners,conditioning regimens, hematopoiesis reconstitution, effectiveness and complication. Results The interval from diagnosis to transplant was 70 (19 - 180) days. Three patients (MRD) underwent BM + PBSCT, one was undergone BM + PBSC + CBSCT. Conditioning regimens of all patients were CY/ATG. Hematopoiesis reconstitution was achieved in 4 patients (100%). The median time of neutrophils which reached 0. 5 x 109/L and platelets reached 20 × 109/L were 14. 5 (9-28) and 16(9 -28) days. Two cases developed grade Ⅰ acute graft-versus-host diseaes (aGVHD), chronic local GVHD occurred in one patient. Four patients are alive with a median time of 40. 6(2 -63) months at the end of the following-up. Conclusions Allo-HSCT are an efficient and safe therapy for the patient with SAA,not only for patients with HLA matched related donor,but also for those only HLA mismatched related donor available.     

造血干细胞移植治疗重型再生障碍性贫血

Objective To evaluate the therapeutic effects of allogeneic hematopoietic stem cell transplantat (allo-HSCT) for severe aplastic anemia (SAA). Methods Four patients of SAA underwent allo-HSCT at the bonemarrow transplant unit in our hospital from March 2003 to May 2009. Stem cell source was an HLA (human leukocyte antigen) matched related donor (MRD) in 3, HLA 1 (B) mismatched related donor in 1 patient A retrospective analysis was performed on interval from diagnosis to transplant,HSCT manners,conditioning regimens, hematopoiesis reconstitution, effectiveness and complication. Results The interval from diagnosis to transplant was 70 (19 - 180) days. Three patients (MRD) underwent BM + PBSCT, one was undergone BM + PBSC + CBSCT. Conditioning regimens of all patients were CY/ATG. Hematopoiesis reconstitution was achieved in 4 patients (100%). The median time of neutrophils which reached 0. 5 x 109/L and platelets reached 20 × 109/L were 14. 5 (9-28) and 16(9 -28) days. Two cases developed grade Ⅰ acute graft-versus-host diseaes (aGVHD), chronic local GVHD occurred in one patient. Four patients are alive with a median time of 40. 6(2 -63) months at the end of the following-up. Conclusions Allo-HSCT are an efficient and safe therapy for the patient with SAA,not only for patients with HLA matched related donor,but also for those only HLA mismatched related donor available.     

脉冲高容量血液滤过对脓毒症患者细胞免疫功能的影响

目的 探讨脉冲高容量血液滤过对脓毒症患者外周血辅助性T细胞(T helper,Th)17及CD4+ CD25+ 调节性T细胞(Treg)影响及其临床价值.方法 本研究为前瞻性对照研究,将2008年1月至2010年11月在安徽省立医院ICU住院的脓毒症患者40例(男/女=24/16),年龄25～75岁,按照疾病严重程度分为3组:脓毒症组14例(男/女=8/6);严重脓毒症组15例(男/女=9/6);脓毒症休克组11例(7/4).入选和疾病严重程度分级标准:根据1992年美国胸科医师学院(ACCP)/美国危重病医学会(SCCM)共识会议制定的脓毒症诊断标准.排除标准:患有自身免疫系统疾病、急性脑卒中、心肌梗死、病毒性肝炎、HIV感染的患者以及入院前3个月内使用过激素或免疫抑制剂的患者.其中入组5 d内未行血液净化治疗患者15例(男/女=8/7)入选为A组;5 d内行脉冲高容量血液滤过的25例患者(男/女=16/9)入选为B组.两组一般资料具有可比性.连续性血液净化以24 h为1周期,两次血滤之间间隔24 h.其中高容量血液滤过(70 mL·kg-1·h-1)治疗6～8 h后续行常规CVVH治疗16～18 h剂量(35 mL·kg-1·h-1).所有入选的40例脓毒症患者在入选当天和第5天清晨空腹抽外周血送检,行流式细胞术检测血中Th17细胞及CD4+ CD25+调节性T细胞的比例.计量资料采用t检验,配对t检验和One way ANOVA分析.小样本率的比较采用确切概率法.相关分析采用Peason相关分析.另选取本院体检中心的20例健康人为健康对照组.结果 健康对照组Th17表达率为(0.91±0.38)%,CD4+ CD25+ Treg细胞表达率为(0.39±0.23)%.40例脓毒症患者在第1天这两项指标明显升高(P<0.05):其中脓毒症组分别为(2.09±0.53)%,(1.72±0.59)%;严重脓毒症组(3.90±0.80)%,(2.72±0.22)%;脓毒性休克组(1.85±0.35)%,(3.55±0.51)%.Th17表达率,严重脓毒症组最高(P<0.05).而脓毒症休克组与脓毒症组比较差异无统计学意义(P>0.05).CD4+ CD25+ 调节性T细胞表达率则呈现:脓毒症休克组>严重脓毒症组>脓毒症组(P<0.05).B组与A组比较,脉冲高容量血液滤过能显著的下调脓毒症患者Th17[(1.87±0.43)vs.(2.48±1.05),P<0.05]和CD4+ CD25+ 调节性T细胞[(1.92±0.89)vs.(2.63±0.92),P<0.05]的表达.结论 脓毒症患者外周血Th17细胞和CD4+ CD25+调节性T细胞表达增加,提示Th17细胞和CD4+ CD25+调节性T细胞在脓毒症的免疫发病机制中可能起着重要作用.脉冲高容量血液滤过能有效的调整Th17细胞和CD4+ CD25+ 调节性T细胞的表达,可作为脓毒症免疫调节治疗的重要手段之一.

Abstract：

Objective To study the effects of pulse high volume hemofiltration (PHVHF) on the changes of Th17 cells (T helper 17 cells) and CD4 + CD25 + reguratory T cells (Treg cells) in peripheral blood of patients with sepsis and to evaluate the clinical value of this intervention. Methods The patients were included in this prospective study as per the criteria of sepsis set by America Chest Physicians College/America Society for Critic Care Medicine in 1992. The patients were excluded: ① immune system disorder, ② acute stroke, ③ myocardial infarction, ④ virus hepatitis,⑤ human immunodeficiency virus infection, ⑥ under immunosuppressive therapy. Forty patients (24 males, 16 females, aged from 25 to 75years) with sepsis in ICU were enrolled from January. 2008 to November. 2010. According to the severity of disease, the patients were divided into three groups; moderate sepsis group (n = 14, 8 males, 6 females) , severe sepsis group (n = 15, 9 males, 6 females) , and septic shock group (n = 11, 7 males, 4 females). The initially clinical data of three groups were comparable. Twenty healthy individuals served as controls. According to the mode of treatment, forty patients were also divided into two groups: conventional treatment group (group A, n= 15) in which patients were treated without PHVHF within 5 days after admission and trial group (group B, n=25) in which patients were treated with pulsed high volume hemofiltration (PHVHF) within 5 days after admission. In group B, high volume hemofiltration (70 mL · kg-1 · h-1) was given to patients for 6 ～ 8 hours, and then conventional continuous vein - vein hemofiltration (35 mL · kg-1 · h-1) for 16 ~ 18 hours. The total length of period for continuum blood scavenging was 24 hours as one cycle. The interval between two cycles of blood scavenging was 24 hours. The changes of Th17 cells and CD4+ CD25 + Treg cells of 40 patients were detected with flow cytometry on the 1st day and the 5th day after admission. The data were analyzed by using SPSS version 13. 0 software. Measurement data were analyzed with Paired-samples t-test, independent-samples t-test or one way ANOVA . Ratio of small samples was compared with fisher's exact test, and the correlation was analyzed by using Pearson correlation analysis. Results The rates of Th17 cells were( 0.91 ±0.38)%, (2.09 ±0. 53)% , (3.90 ±0. 80)% , and ( 1. 85 ±0.35)% in control, moderate sepsis, severe sepsis, and septic shock groups, respectively, while the rates of CD4+ CD25+ Treg cells were (0.39 ±0.23)%, (1. 72 ±0. 59)% , (2.72 ±0. 22)% , and (3. 55 ±0. 51)% , respectively. The rate of Thl7 cells on the 1st day was higher in severe sepsis group than that in other two groups ( P < 0. 05 ) without significant difference between septic shock and moderate sepsis groups ( P > 0. 05). Moreover , the rate of CD4+ CD25 + Treg cells was up - regulated on the 1st day in the following order from high to low: septic shock group > severe sepsis group > sepsis group (P < 0.05). The rates of Th17 cells and CD4 + CD25 + Treg cells in patients of group B decreased in greater degree than that did in patients of group A (P < 0.05 ). Conclusions The changes of Th17 cells and CD4 + CD25 + Treg cells may play an important role in pathogenesis of sepsis, and the pulsed high volume hemofiltration may be one of the effective treatments for the patients with sepsis by regulating the rates of Thl7 cells and CD4 + CD25 + Treg cells.     

Oncogenic role of Tc17 cells in cervical cancer development

BACKGROUND As one of the subsets of CD8+T cells,Tc17 cells have recently been identified and are characterized by the secretion of interleukin(IL)-17,which is related to inflammatory diseases.AIM To assess the status of Tc17 cells in cervical cancer and investigate the biological function of Tc17 cells in cervical cancer development.METHODS Flow cytometry assay,immunohistochemistry,and immunofluorescence were performed to detect the levels and phenotype of Tc17 cells in blood and tumor samples from patients with cervical cancer.Prior to cell suspension culture,ELISA was carried out to measure the production of IL-6,IL-1β,IL-23,CXCL12,and IL-17 in tumor tissue supernatant and co-cultured supernatant of patients with cervical cancer.In addition,multivariate analysis was performed to identify factors associated with overall survival using the Cox proportional hazards model.RESULTS Compared with normal tissues,Tc17 cells specifically accumulated in tumor tissues of cervical cancer patients.Cancer cells produced a greater amount of IL-6,IL-1β,and IL-23,which in turn promoted Tc17 cell polarization.Unlike the traditional cytotoxic CD8^+T cells,Tc17 cells secreted IL-17,which subsequently promoted CXCL12 expression in tumor cells,eventually enhancing the proliferation and migration of tumor cells.Thus,the ratio of tumor-infiltrating Tc17 cells was highly correlated with poor clinical outcome in patients with cervical cancer.CONCLUSION Our data identified the oncogenic role of Tc17 cells in the development of cervical cancer.We propose that the ratio of Tc17 cells may be a useful index in the prognosis of patients with cervical cancer.     

100例异基因造血干细胞移植后移植物抗宿主病的发生及其对患者复发和生存的影响

本研究探讨异基因造血干细胞移植（allo—HSCT）后急性移植物抗宿主病（aGVHD）和慢性移植物抗宿主病（cGVHD）的危险因素及其与预后的关系。对我院100例行allo—HSCT患者的临床资料进行了总结,对aGVHD、cGVHD的发生率和危险因素及其对复发和生存的影响进行了分析。结果表明：31例患者发生Ⅱ-Ⅳ度aGVHD,累积发生率为34．4％;14例发生Ⅲ-Ⅳ度aGVHD,累积发生率为17．7％。HLA相合程度与aGVHD的发生无显著相关性（P〉0．05）。曾发生的Ⅱ-Ⅳ度aGVHD是发生cGVHD的危险因素（HR=2．303,P=0．088）,女性因素为保护性因素（HR=0．401,P=0．055）。发生cGVHD者复发率较低。发生Ⅱ-Ⅳ度aGVHD为影响总生存率（OS）的危险因素（P〈0．05）,重度（Ⅲ-Ⅳ）aGVHD患者的死亡率极高（81．0％）,与0-Ⅰ度患者的死亡率（35．7％）相比有显著性差异（P=0．000）。结论：GVHD和移植物抗肿瘤效应（GVL）尚不能分离,但GVL的益处可能被GVHD相关死亡因素抵消,因此需积极防治重度aGVHD。局限型cGVHD可能是长期生存的有益因素。     

抑制性和激活性免疫球蛋白样受体在亲缘半相合造血干细胞移植中的作用

目的 分别探讨NK细胞中抑制性和激活性免疫球蛋白样受体(KIR)在亲缘半相合造血干细胞移植(HSCT)中的作用.方法 检测47例亲缘半相合HSCT中的供者KIR基因型和患者HLA-C基因型,随访移植后2年总生存率、Ⅲ～Ⅳ度急性移植物抗宿主病(GVHD)发生率和复发率.结果 ①按供受者抑制性KIR与其配体HLA-C是否匹配分为两组,匹配组的2年总生存率明显高于不匹配组[分别为(87.5±8.3)%和(54.5±9.0)%,P=0.03];复发率明显降低[分别为0和(25.4±9.5)%,P=0.05].将病例分为髓系白血病和淋巴细胞白血病,其中30例髓系白血病患者中匹配组复发率明显低于不匹配组[分别为0和(35.0±14.4)%,P=0.04].其他项目差异无统计学意义.②常见的激活性KIR主要包括3个:KIR2DS1、KIR2DS2、KIR2DS3.分别按其供者是否表达进行单因素分析,其中KIR2DS1(+)组Ⅲ～Ⅳ度急性GVHD发生率比KITR2DS1(-)组低(分别为13%和28%,P＞0.05);KIR2DS2(+)组复发率低于KIR2DS2(-)组[分别为0和(17.3±7.1)%,P＞0.05];KIR2DS3(+)组Ⅲ～Ⅳ度急性GVHD发生率低于KIR2DS3(-)组(分别为11%和26%,P＞0.05);其他无明显差异.结论 ①亲缘半相合HSCT中,供者抑制性KIR与患者HLA-C匹配组较不匹配组总体生存率明显升高,复发率明显降低.特别在髓系白血病中,复发率明显降低.②对于激活性KIR受体中KIR2DS1或KIR2DS3的表达,可能降低急性重度GVHD的发生率;而KIR2DS2(+)组表达,可能降低复发率.

Abstract：

Objective To investigate the effect of inhibitory and activating KIRs on a cohort of Chinese leukemia patients who received haplo-identical hematopoietic stem cell transplantation (HSCT).Methods Donor' s inhibitory and activating KIRs and recipient' s HLA-C from 47 cases who received haplo-identical HSCT were tested by PCR-SSP.2 year overall survival ( OS), incidence of severe ( grade Ⅲ to Ⅳ )acute GVHD (aGVHD) and relapse rate(RR) were analyzed.Results ①According to Matched (M) vs Mis-Matched( MM ) between donor' s inhibitory KIR and recipient' s HLA-C1/C2 subgroup, 2 year OS rate in M group[(87.5 ±8.3)%]was significantly higher than that in MM group (54.5 ±9.0)%, (P=0.03).Lower incidence of relapse rate was seen in M group than in M/MM groups[0 vs (25.4 ±9.5)%, P =0.05].In 30 cases of myeloid leukemia patients, there was lower RR in M group than in MM groups[0 vs (35.0 ± 14.4) % ,P = 0.04].②According to the 3 activating KIR genes: KIR2DSI/KIR2DS2/KIR2DS3,lower incidence of grade Ⅲ - Ⅳ aGVHD was seen in KIR2DS1 ( + ) group than in KIR2DS1 ( - ) group ( 13% vs 28%, respectively, P＞0.05); and so was done in KIR2DS3( + ) group( 11% vs 26%, respectively, P＞0.05).The RR was lower in KIR2DS2( + ) group[0% vs (17.3 +7.1)%, respectively, P＞0.05].Conclusions In our haplo-identical HSCT setting, match between donor' s inhibitory KIR and recipient's HLA-C can significantly reduce the incidence of relapse rate and improve OS.Although lower incidences of severe aGVHD are noted in the donors with KIR2DS1 ( + ) or KIR2DS3 ( + ), and lower relapse rate is noted in the donors with KIR2DS2( + ) but without statistic difference, no remarkable effects of activating KIRs on OS have been found in our relatively small clinical series.     

异基因外周血干细胞移植治疗白血病中移植物抗宿主病的研究

目的　分析异基因外周血造血干细胞移植 (allo PBSCT)治疗白血病过程中移植物抗宿主病 (GVHD)的发生状况 ,探讨其有效防治方法。方法　allo PBSCT治疗白血病 5 0例 (急性 2 9例 ,慢性 2 1例 ) ;预处理采用环磷酰胺 (CTX) +足叶乙甙 (Vp16 ) +全身照射 (TBI)或CTX +TBI方案 ;32例患者GVHD预防采用常规环孢菌素A(CsA) +短程甲氨蝶呤 (MTX)方案 ;18例患者采用霉酚酸酯 (MMF)联合CsA +短程MTX方案。结果　患者经allo PBSCT均获得造血功能重建 ,中性粒细胞 >0 .5× 10 9 L和血小板 >2 0× 10 9 L时间分别为移植后 14 (10～ 2 2 )d和 2 0 (10～ 6 8)d。发生急性GVHD(aGVHD) 2 0例(40 % ) ,其中Ⅲ～Ⅳ度 6例 (12 % )。生存 6个月以上可评价的 33例中 2 2例 (6 6 7% )发生慢性GVHD(cGVHD) ,其中广泛性 11例 (33.3% )。aGVHD阳性组可溶性白细胞介素 2受体 (sIL 2R)水平为 (2 77.3± 2 6 .4 )U L ,CD2 5+ 细胞为 (8.1± 3.4 ) % ,明显高于aGVHD阴性组 [分别为 (12 8.1± 96 .7)U L和 (3.6±1.7) % ](P <0 .0 5 )。采用MMF联合CsA +MTX方案组aGVHD和广泛性cGVHD发生率分别为 16 .7%和 9.1% ,明显低于常规CsA +MTX方案组 (分别为 5 3.1%和 4 5 .5 % ) (P <0 .0 5 )。本组患者中位随访30 (3～ 70 )个月 ,GVHD阳性组     

Clinical study on the relapsd leukemia patients with graft versus host disease after allogeneic hematopoietic stem cell transplantation

OBJECTIVE: To explore the dissociation of graft-versus-leukemia (GVL) effects from graft-versus-host disease (GVHD) in the patients who experienced GVHD during leukemia relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT). METHODS: The primary disease, disease status, GVHD, response to donor lymphocyte infusion (DLI) and prognosis were analysed in 11 leukemia patients who relapsed with GVHD after allo-HSCT. RESULTS: Of the 11 relapsed, 5 were acute lymphoblastic leukemia and 6 acute myeloid leukemia. Five received DLI before relapse and all developed post-DLI GVHD, including 2 grade II acute GVHD (aGVHD), 1 limited chronic GVHD (cGVHD) plus grade II aGVHD, and 2 extensive cGVHD. After relapse of the 5 patients, 2 received Chemo-DLI, one achieved CR with extensive cGVHD and then relapsed again, the other didn't achieved CR. The other 6 patients didn't received DLI before relapse and also developed post-HSCT GVHD while relapsing, including 3 extensive cGVHD, 1 grade I aGVHD and 2 grade II-IV aGVHD. After relapse, these 6 patients received Chemo-DLI, 2 achieved CR and then relapsed again, 4 didn't achieved CR. CONCLUSION: The elicited GVHD after allo-HSCT may not accompany effective GVL effects inhibiting leukemic relapse.     

Impact of donor and recipient characteristics on graft-versus-host disease and survival in HLA-matched sibling hematopoietic stem cell transplantation

ObjectiveTo analyze the impact of donor- and recipient-related factors on Graft-versus-host disease (GVHD) and overall survival of transplantation from matched sibling donors.Methodwe retrospectively analyzed the clinical features of 68 consecutive hematological patients undergoing allogeneic hematopoietic stem cell transplantation (allo-HSCT) from matched sibling from 2011 and 2017.ResultsThe incidence of Ⅱ- Ⅳacute GVHD (aGVHD) and chronic GVHD (cGVHD) after transplantation was 13.6 % and 19.7 %, respectively. We also noted the donor and recipient characteristics had no impact on Ⅱ- Ⅳ aGVHD incidence.We found sex mismatch (F-M) did not increase the risk of cGVHD in the model if a female donor was younger than 30 years (P = 1.000), but cGVHD increased if the female donor was ≥30 years (P = 0.002). Recipients≥40 years undergoing HCT from donors ≥30 years were at higher risk for cGVHD (P = 0.021). Development of Ⅱ- Ⅳ aGVHD and cGVHD had no effect on overall survival (P = 0.159, 0.081). Non-remission status at allo-HCT was linked to lower overall survival (P = 0.001).ConclusionThe incidence of cGVHD was higher when male recipients received hematopoietic progenitor cells from female ≥30 years donors, and when older than 40 years recipients received hematopoietic progenitor cells from ≥30 years donors. Patients in non-remission status at allo-HCT was inclined to have lower overall survival.     

受者特异性产生干扰素γ细胞与急性移植物抗宿主病的关系

目的研究异基因造血干细胞移植(allo-HSCT)中针对受者特异性产生干扰素γ细胞 (IFN-γ PC)与急性移植物抗宿主病(aGVHD)的关系。方法以37例HLA相合的同胞allo-HSCT患者为研究对象,将移植前供者、移植后患者外周血单个核细胞(PBMNC)分别作为反应细胞(RC),移植前受者PBMNC作为异基因刺激细胞(allo-SC),在混合淋巴细胞反应后用酶联免疫斑点法检测针对受者特异性IFN-γ PC,分析其与aGVHD的关系。结果 allo-HSCT前检测受者特异性IFN-γ PC水平, 移植后发生aGVHD组显著高于无aGVHD组(P<0．01);供者针对受者特异性IFN-γ Pc≥20／2×10~5 RC时,发生Ⅱ-Ⅳ度aGVHD的比例显著高于<20／2×10~5RC组(P<0．05)。aGVHD发生时患者针对移植前受者特异性IFN-γ PC水平显著高于移植前相应供者针对受者的水平(P<0．05);亦显著高于 aGVHD患者针对移植前供者的水平(P<0．01)。移植后14天(+14天)和+28天时,aGVHD组受者特异性IFN-γ PC水平均显著高于无aGVHD组(P值均<0．01)。aGVHD患者经过免疫抑制剂治疗7 d后,受者特异性IFN-γ PC较aGVHD发生时显著降低(P<0．05)。结论受者特异性IFN-γ PC的水平能够反映allo-HSCT前后的同种异体反应,有助于临床aGVHD的预测、诊断和监测。     

非血缘关系异基因造血干细胞移植66例分析

目的对66例血液病患者进行非血缘关系异基因造血干细胞移植（allo-HSCT），探索提高移植疗效的措施。方法慢性粒细胞白血病（CML）患者24例，急性白血病（AL）患者40例，其他血液病患者2例，经预处理治疗后，进行人类白细胞抗原（HLA）基本相合的非血缘关系骨髓移植（BMT）48例，外周血干细胞移植（PBSCT）18例：部分患者采用长程加强的移植物抗宿主病（GVHD）的预防方案（将环孢菌素A提前至预处理开始时使用，同时加用霉酚酸酯）。结果64例患者达到完全稳定的供者植入，WBC植活中位时间15d（BMT组16d；PBSCT组12d，P〈0．01）。45例患者发生急性GVHD（aGVHD），累积发生率为71．16％，其中28例患者发生Ⅰ～Ⅱ度GVHD，累积发生率57．15％；17例患者发生Ⅲ～Ⅳ度GVHD，累积发生率32．25％；COX模型分析得出HLA配型及移植方式是影响aGVHD发生的因素，HLA配型相合、采用G-CSF动员的PBSCT可以降低aGVHD，尤其是重度GVHD的发生。可供分析的36例患者中有21例发生慢性GVHD。66例接受移植的患者中复发6例，死亡27例，5年的预期生存率为52．91％。用COX模型分析得出aGVHD以及aGVHD与GVHD的预防方案的交互冈素是影响生存率的惟一因素，其相对危险度分别为1.517和1.255。结论提高非血缘关系allo-HSCT疗效的关键是控制aGVHD，而选择HLA配型相合的供者，加强移植早期的免疫抑制，可以减少aGVHD的发生     

无血缘关系供者外周血干细胞移植和骨髓移植治疗白血病的比较

目的比较无血缘关系供者外周血干细胞移植和骨髓移植在造血重建、T 细胞重建、感染、移植物抗宿主病(GVHD)及疗效等的差异。方法 53例白血病患者中,21例接受无血缘关系供者外周血干细胞移植(PBSCT 组),32例接受无血缘关系供者骨髓移植(BMT 组)。统计分析二者移植后白细胞和血小板重建时间、T 细胞重建、感染率、GVHD、白血病复发、无病生存(DFS)情况。结果PBSCT 组和 BMT 组移植后白细胞重建时间分别为(12.43±3.67)天和(16.16±2.99)天(P<0.01),血小板重建时间分别为(14.67±6.19)天和(21.23±8.25)天(P<0.01)。PBSCT 组和 BMT 组移植后1,3,6,9及12个月的 T 细胞重建差异无统计学意义。PBSCT 组和 BMT 组移植后早期感染率分别为42.86%和53.13%(P>0.05)。PBSCT 组与 BMT 组急性 GVHD 的发生率分别为61.90% 和71.885(P>0.05);在可统计的患者中,慢性 GVHD 的发生率在 PBSCT 组和 BMT 组分别为47.06%和43.48%(P>0.05)。PBSCT 组与 BMT 组移植后分别有4例和2例复发,二者复发率差异无统计学意义(P>0.05);PBSCT 组与 BMT 组移植后2年 DFS 率分别为(50.14±12.00)% 和(59.81±8.99)%,二者差异也无统计学意义(P>0.05)。结论无血缘关系供者 PBSCT 后的造血重建比BMT,但二者移植后 T 细胞重建、感染发生率、GVHD 及 DFS 的差异均无统计学意义。     

Ly49A转基因小鼠脾细胞对半相合异基因骨髓移植后GVHD和GVL的作用

目的　观察Ly4 9A基因转染的C5 7BL 6小鼠脾细胞对半相合异基因骨髓移植 (allo BMT)后移植物抗宿主病 (GVHD)和移植物抗白血病效应 (GVL)的影响。方法　经由逆转录病毒介导将Ly4 9A基因转染至C5 7BL 6小鼠的脾细胞 ;流式细胞仪检测Ly4 9A受体在转染后脾细胞上的表达率 ;以亲代C5 7BL 6H 2 b 小鼠为供者 ,以接种EL96 11红白血病细胞的 (BALB c×C5 7BL 6 )F1H 2 d b(CB6F1)小鼠为受者 ,预处理条件为全身照射 (TBI,6 0 Co照射 10 .5Gy) ,进行脾细胞混合骨髓细胞移植 ,建立半相合异基因急性GVHD模型并在该模型基础上观察Ly4 9A基因转染的脾细胞对GVHD和GVL的作用。结果　Ly4 9A基因转染的C5 7BL 6小鼠的脾细胞 2 4h后蛋白表达率为 (42 .2 0± 4 .87) % ,空载体转染组为(18.6 7± 2 4 8) % ,未转染对照组为 (18.73± 3.82 ) % ,在半相合异基因移植中 (C5 7BL 6 H 2b →CB6F1H 2d b) ,未进行移植的单纯照射组生存期为 (7.80± 3.36 )d ;环磷酰胺治疗组生存期为 (2 1.70±2 87)d ;脾细胞混合骨髓细胞移植组生存期为 (2 9.4 0± 6 .4 3)d ;空载体转染的脾细胞混合骨髓细胞移植组生存期为 (2 9.10± 7.39)d ;Ly4 9A转染的脾细胞混合骨髓细胞移植组生存期为 (45 .0 0± 12 .38)d ,较上述各组生存期明显延长 (P