Serial assessment of circulating T lymphocyte phenotype and receptor repertoire during treatment of non-muscle invasive bladder cancer with adoptive T cell immunotherapy

Recurrence and progression of non-muscle-invasive bladder cancer (NMIBC), frequent despite the availability of multiple treatment modalities, may be partly explained by the presence of immunosuppressive cell populations. We hypothesized that progression of disease could be prevented by the administration of an activated T cell immunotherapy (ACT) at time points when immunosuppressive populations increased in peripheral blood. In an N-of-1 study, a patient with multiple primary bladder high grade urothelial carcinomas, previously treated with standard local resection and chemotherapy but with evidence of progression, received ACT consisting of dendritic cells mixed with cytokine induced killer cells (DC/CIK), intravenously 18 times over a 6 year period at indicated time of observed increases in peripheral blood immunosuppressive CD8⁺/CD28^- cells. Peripheral blood was analyzed for T cell phenotype by flow cytometry, T cell receptor (TCR) repertoire, and circulating tumor DNA (ctDNA) by next generation sequencing (NGS) at the time of each infusion. Cystoscopy and pelvic CT scans were performed at routine intervals to assess clinical status of disease. There has been no recurrence or metastasis of urothelial carcinoma. Peripheral blood cytotoxic T cells and unique TCR clones increased and suppressive T cell populations decreased after DC/CIK infusions evidenced by the two more proof-of concept cases. ctDNA analysis detected mutations in six genes (ARID1B, MYCN, CDH23, SETD2, NOTCH4 and FAT1) which appeared at different times, but all of them disappeared after the DC-CIK infusions. These data suggest that DC/CIK infusions may be associated with beneficial changes in T cell phenotype, TCR repertoire, decreases in circulating tumor DNA and sustained recurrence-free survival.     

Oligoclonal T cells in human cancer

Many solid tumors are characterised by the infiltration of lymphocytes and their presence has been correlated with a more favourable prognosis. These tumor-infiltrating lymphocytes (TIL), have been shown to possess specific cytolytic reactivity towards autologous tumours, thus suggesting that tumour cells may express antigens capable of eliciting an immune response. Expression of such tumour-associated antigens (TAA) in combination with appropriate accessory signals would lead to thein vivo accumulation of T cells with anti-tumour specificity. Analysis of the composition of the specific T-cell receptor (TCR) of TIL could thus provide information on the nature of the antigen(s) recognised by TIL. In this review, different aspects of the presence of clonal T cells in patients with cancer are discussed.     

Increased apoptosis of circulating T cells in myelodysplastic syndromes

The mechanism of T cell lymphopenia in myelodysplastic syndromes (MDS) is unknown. We investigated apoptosis in freshly isolated and cultured lymphocytes; the latter were used to detect cells not yet apoptotic but destined for apoptosis. Apoptosis increased in both fresh and cultured T cells in MDS compared with those from healthy controls. Furthermore, in lymphopenic MDS patients the lymphocyte count correlated negatively with the degree of T cell apoptosis. MDS T cells showed increased Fas expression. However, in MDS but not in controls, the degree of T cell apoptosis was independent of the Fas expression level, and exogenous anti-Fas antibodies did not modulate T cell apoptosis. Mechanisms other than the Fas–Fas ligand pathway may induce T cell apoptosis in MDS.     

Treatment of myelodysplastic syndromes

The IPSS scoring system is useful to establish the appropriate treatment plan in MDS. Growth factors may alleviate both anemia and neutropenia in some MDS patients. Serum Epo levels and need for transfusion serve as good predictors of the erythroid response to the combination of Epo and G-CSF. Subgroups of MDS patients may respond favorably to immunosuppressive therapies such as CyA and ATG. Low-dose chemotherapy may also improve peripheral blood counts. Platelet counts, bone marrow cellularity, chromosome aberrations, and ringed sideroblasts combine to create a model predicting the response to low-dose ara-C. High-dose chemotherapy may lead to complete remission in about half of MDS patients, but the duration of remission is often short. The only proven curative therapy for MDS is allogeneic stem cell transplantation, resulting in an overall disease-free survival rate of about 40%. Only a minority of patients, however, can undergo allogeneic transplantation, both because of patient age and the availability of suitable donors. Autologous stem cell transplantation may be an option for selected patients who are unable to find allogeneic donors. Because the clinical features of patients with MDS are quite heterogeneous, the development of more accurate predictive models may be necessary to improve the efficacy of treatment.     

Prognostic implication and characterization of the blast cell population in the myelodysplastic syndrome

The bone marrow smears of 18 confirmed cases of MDS were analyzed carefully for the presence of “hypergranular type III blasts”, defined as more than 20 fine azurophil primary granules per cells. The concordance was close to 80% among 5 observers. Thirty-nine percent (7 cases) were reclassified as RAEB-t rather than RAEB. The presence of these hypergranular blasts was not suggestive of increased differentiation but rather leukemic cells. The reassignment of cases altered the median survival for the various subcategories, providing a clearer separation with the introduction of type III blasts than without utilizing these cells which were separated from the promyelocyte family. The introduction of this new blast cell definition in a larger series of patients is recommended to confirm these preliminary observations.     

骨髓增生异常综合征免疫表型特征及其临床意义

 骨髓增生异常综合征（MDS）患者骨髓细胞免疫表型在整个疾病过程中呈现出紊乱及异常表达,其中一些改变对其诊断、分型、预后和治疗等方面有一定的价值。就此方面的进展作一综述。     

干细胞移植治疗骨髓增生异常综合征13例

 目的　探讨干细胞移植（HSCT）在骨髓增生异常综合征（MDS）治疗中的疗效以及预处理方式的选择。方法　对13例MDS患者行异基因造血干细胞移植 （allo-HSCT）（包括HLA配型全相合10例、半相合2例、脐血移植1例）。输注单个核细胞（MNC） 6.92（2.65～21.33）×108／kg,CD34细胞4.47（1.49～10.22）×106／kg。其中,5例选择全身照射+氟达拉滨+环磷酰胺（TBI+Flud+Cy）方案预处理,3例白消安（BU）／Cy预处理,3例TBI+CY,2例采用阿糖胞苷（Ara-C）+BU+Cy+替尼泊苷（VM26）预处理。移植物抗宿主病（GVHD）的预防：2例HLA配型半相合者给予抗胸腺细胞球蛋白（ATG）联合环孢素A（CsA）加短程甲氨蝶呤（MTX）治疗,并于移植后1～28 d持续给予霉酚酸酯（MMF）,其他病例仅给予CsA加短程MTX。结果　13例患者中9例造血完全重建,移植相关死亡4例。结论　HSCT是可以治愈MDS的有效方法。预处理选择应采取个体化。     

异基因造血干细胞移植治疗骨髓增生异常综合征:适应证及移植前是否需化疗

 异基因造血干细胞移植（allo-HSCT）治疗骨髓增生异常综合征（MDS）的适应证以FAB分型、国际预后评分系统（IPSS）、世界卫生组织预后评分系统（WPSS）为基础，IPSS中危-2及高危的患者应尽可能在诊断早期接受移植; 中危-1及以下者将从延迟移植中受益。临床决策中应动态随诊病情变化、及时把握疾病状态及进展速度适时移植。中低危患者血小板极低、中性粒细胞缺乏或输血依赖重者应尽早移植。MDS中高危患者移植前是否该应用去甲基化药物或化疗尚存争议，迄今临床研究数据表明移植前化疗对移植预后无益。     

Intensive chemotherapy for acute non-lymphoblastic leukemia after primary myelodysplastic syndrome

Twenty-five patients with a primary myelodysplastic syndrome (MDS) transformed into acute non-lymphoblastic leukaemia (ANL) were treated with intensive chemotherapy. A complete remission (CR) was obtained in six patients (24 per cent). In five of these six patients two courses of chemotherapy were needed to achieve CR. In eight patients chemotherapy cleared the bone marrow of blasts, but the aplasia was fatal. A partial effect on bone marrow blasts was seen in four patients and no effect in another six. Eleven patients (44 per cent) died from the consequences of chemotherapy-induced cytopenia. A short interval between MDS and transformation into ANL was associated with a better chance of achieving complete remission. Age, karyotype, type of MDS, peripheral blood or bone marrow findings had no influence on the result of chemotherapy. The median survival from start of treatment was 5 months (range 0.5-24 months). In the patients who achieved a CR, the median duration of the remission was 7 months (range 3-12 months). The poor response rate, the short duration of the remissions and the high treatment-related mortality suggest that current intensive anti-leukemic chemotherapy in ANL after primary MDS is of limited benefit.     

骨髓增生异常综合征患者染色体核型分析

 目的　对骨髓增生异常综合征（MDS）染色体核型进行分析并结合血细胞计数、骨髓原始细胞数对其预后进行评估。方法　采用直接法、短期培养法和反带技术制备染色体,进行核型分析。结果　49例MDS患者中有22例（44.9 %）检出异常克隆。核型异常包括数目异常和结构异常,数目异常以-7,+8最常见。难治性贫血伴原始细胞增多（RAEB1和RAEB2）较难治性贫血（RA）和难治性贫血伴有环状铁粒幼细胞（RARS）检测到更高的异常核型比例。MDS染色体核型异常与原始细胞比例呈正相关。结论　染色体核型分析对MDS诊断、治疗及预后评估有重要价值。     

Reduced-intensity conditioning therapy with fludarabine,idarubicin, busulfan and cytarabine for allogeneic hematopoietic stem cell transplantation in acute myeloid leukemia and myelodysplastic syndrome

We retrospectively analyzed allogenic stem cell transplantation (allo-SCT) outcomes in 82 patients with AML or MDS were conditioned with fludarabine, idarubicin, intravenous-busulfan and cytarabine (FIBA) or busulfan and cyclophosphamide (BuCy). Compared to BuCy regimen, reduced intensity conditioning (RIC) with FIBA was associated with a lower incidence of severe acute GVHD, lower NRM and a similar relapse rate. There was no significant difference in the 3 year overall survival (OS), but this is possibly due to the limited number of patients. The FIBA regimen is promising to replace BuCy regimen because of better security and similar relapse rate.     

Epidemiological data from the registry of patients with myelodysplastic syndrome in a single hospital center of Romania

We present the first Romanian study on the epidemiological characteristics of MDS, based on the data existing in Fundeni Clinical Institute, Hematological Department, Bucharest. The files at diagnosis of the adult patients with primary MDS admitted during the period 1982–2005, recorded in the registration forms provided by the MDS Foundation (USA), represented the primary database. This study indicates an increase in the number of new MDS cases over the period of time investigated. Also, a 10 years lower median age of the patients, a noticeable proportion of young patients and a low proportion of patients ≥81years have been found, which situates our findings in the middle between the Eastern and Western epidemiological reported data on MDS.     

骨髓增生异常综合征和再生障碍性贫血患者骨髓CD34+细胞测定

 目的 检测骨髓增生异常综合征（MDS）和再生障碍性贫血（AA）患者骨髓CD+34细胞占单个核细胞（MNC）的比率,以探讨二者可能的发病机制。方法 用流式细胞术（FCM）检测22例MDS患者、13例AA患者及12例非血液病患者骨髓CD+34细胞占MNC的比率。结果 AA组与对照组、AA组与MDS-RA组、AA组与MDS-RAEB组、MDS-RA组与MDS-RAEB组的骨髓MNC中CD+34细胞的比率的比较差异有统计学意义（P＜0.05）。大多数重型AA（SAA）患者（3／4）及很少慢性AA（CAA）患者（1／9）的骨髓MNC中的CD+34细胞的比率＜0.1 %。结论 骨髓CD+34细胞的检测有助于判断AA患者病情及MDS患者的预后,亦可用于鉴别AA和MDS。     

造血干细胞移植治疗骨髓增生异常综合征的研究进展

 人类白细胞抗原（HLA）匹配同胞供者造血干细胞移植是治疗骨髓增生异常综合征（MDS）最有效的方法,其疗效在过去十几年里有明显的提高。在缺乏同胞供者来源时,无关供者移植、脐带血移植、自体造血干细胞移植可以作为替代的治疗选择。现对MDS患者移植适应证、影响移植疗效的因素（包括年龄、移植时机、移植前诱导缓解化疗、预处理方案、造血干细胞来源等）进行综述。     

Cytogenetic characteristics and prognosis analysis in 231 myelodysplastic syndrome patients from a single institution

We analyzed the clinical and hematologic data of 231 patients diagnosed with de novo myelodysplastic syndrome (MDS), identified cytogenetic characteristics, and evaluated the significance of prognostic systems. The median age was 51 years and the distribution of MDS subtypes demonstrated a markedly low incidence of MDS with deletion 5q (0.9%). The proportions of World Health Organization (WHO) categories differed according to patient age group. Refractory anemia with excess blasts-2 demonstrated the most significant trend toward increased frequency with advancing age. The incidence of abnormal karyotypes in our study was comparable to a previous study (50.2%), although with different patterns. The most frequent cytogenetic abnormality was +8 (34.5% of patients with abnormality), followed by 1q+ (17.2%), 5q− (15.5%), and 20q− (12.9%). Majority of +8, 1q+, −5/5q− and −7/7q− cases combined with additional cytogenetic abnormalities (60.0%, 75.0%, 88.5% and 100%, respectively). The median survival time was 49.5 months and 13.8% patients developed acute leukemia. WHO Prognostic Scoring System (WPSS) and age group were significant factors associated with overall survival. Otherwise, International Prognostic Scoring System was not included in the model. These results demonstrated the different cytogenetic features in Korean MDS patients compared to those of Western country. In addition, WPSS and age group are applicable to our patients as an effective and reliable prognostic model.     

骨髓增生异常综合征对白血病的诊断价值分析

目的探讨骨髓增生异常综合征对白血病的诊断价值。方法选取2010年2月至2013年9月间收治的急性髓系白血病患者120例作为观察组,选择同期进行健康体检者120例作为对照组,两组均进行血常规检测与细胞形态学分析,同时抽取骨髓进行骨髓免疫检测和骨髓增生程度判定。结果观察组患者的白细胞、红细胞与血小板计数均明显低于对照组,差异有统计学意义(P<0.05)。观察组患者的细胞形态学主要表现为细胞大小明显不均匀,核形态分叶过多、形态异常,染色质网状排列等。观察组患者的CD+34CD+123/CD+34比例为(38.24±14.74)%,而对照组为(1.90±0.89)%,差异有统计学意义(P相似文献   

The efficacy of unrelated cord blood transplantation for adult myelodysplastic syndrome

Although allogeneic stem cell transplantation from a human leukocyte antigen (HLA)-identical related donor offers a potential cure for patients with myelodysplastic syndrome (MDS), a suitably matched related donor is unavailable for approximately two thirds of patients. Recently, umbilical cord blood from unrelated donors have been used as an alternative stem cell source for adult patients with MDS. Here, we updated the results of unrelated cord blood transplantation (CBT) after myeloablative conditioning for 22 adult patients with MDS. Diagnosis at transplantation included refractory anemia (RA) (n = 3), refractory anemia with excess blasts (RAEB) (n = 2), RAEB-t (n = 2), and MDS-related secondary acute myeloid leukemia (AML) (n = 15). All patients were treated with total body irradiation (12 Gy), cytosine arabinoside (Ara-C) and cyclophosphamide followed by unrelated HLA-mismatched CBT. The median age was 40 years (range, 19 - 51 years), the median weight was 54.5 kg (range, 43 - 75 kg), and the median number of cryopreserved nucleated cells was 2.43 × 10⁷/kg (range, 1.82 - 4.10 × 10⁷/kg). Twenty one patients had myeloid reconstitution and the median time to more than 0.5 × 10⁹/l absolute neutrophil count was 22.5 days. A self-sustained platelet count more than 50 × 10⁹/l was achieved in 19 patients at a median time of 49 days. Acute GVHD above grade II occurred in seven of 21 evaluable patients and chronic GVHD in 16 of 19 evaluable patients. Among 16 chronic GVHD patients, in eight patients the disease was extensive. Seventeen patients are alive and free of disease at between 371 and 2562 days after transplantation. With a median follow-up of 1505 days, the probability of disease-free survival at 4 years was 76.0%. These results suggest that adult MDS patients without suitable related or unrelated bone marrow donors should be considered as candidates for CBT.     

丙戊酸钠联合地西他滨治疗骨髓增生异常综合征疗效分析

目的 探讨丙戊酸钠联合地西他滨治疗骨髓增生异常综合征(MDS)的有效性和安全性.方法 选择2012年2月至2017年2月山西大医院血液科收治的42例MDS患者为研究对象.将患者按照随机数字表法分为对照组(21例)和试验组(21例),对照组接受地西他滨治疗,剂量20 mg·m-2·d-1,2 h完成静脉滴注,连续治疗5 d,4周为1个疗程;试验组在使用地西他滨治疗的基础上,口服丙戊酸钠0.2 g/次,3次/d,1周后加量至0.4 g/次,3次/d.两组均至少进行4个疗程的治疗.出现严重的不良反应或疾病明显进展时停用,治疗后每4周复查一次骨髓涂片,评价疗效.并分别于治疗前后用荧光定量聚合酶链反应检测患者骨髓细胞中ASXL1、DNMT3A、TET2的表达情况.结果 试验组与对照组的总体治疗反应率分别为76.2%(16/21)和57.1%(12/21),差异有统计学意义(P<0.05);两组的总缓解率分别为47.6%(10/21)和38.1%(8/21),差异无统计学意义(P>0.05);两组患者均有轻微的药物不良反应,不良反应发生率分别为42.9%(9/21)和38.1%(8/21),差异无统计学意义(P>0.05);治疗后两组TET2 mRNA、DNMT3A mRNA含量较治疗前下降,差异均具有统计学意义(P<0.05),但两组间治疗后比较差异无统计学意义(P>0.05);对照组ASXL1 mRNA含量较治疗前无显著变化,试验组患者的ASXL1 mRNA含量较治疗前下降,差异有统计学意义(P<0.05).结论 丙戊酸钠联合地西他滨治疗MDS效果良好,不良反应轻微,且对MDS相关基因TET2、DNMT3A以及ASXL1的表达均有影响.     

Immune correlates of clinical benefit in a phase I study of hyperthermia with adoptive T cell immunotherapy in patients with solid tumors

Abstract

Purpose: To characterize the T cell receptor (TCR) repertoire, serum cytokine levels, peripheral blood T lymphocyte populations, safety, and clinical efficacy of hyperthermia (HT) combined with autologous adoptive cell therapy (ACT) and either salvage chemotherapy (CT) or anti-PD-1 antibody in patients with previously treated advanced solid tumors.

Materials and methods: Thirty-three (33) patients with ovarian, pancreatic, gastric, colorectal, cervical, or endometrial cancer were recruited into the following therapeutic groups: HT?+?ACT (n?=?10), HT?+?ACT?+?anti-PD-1 inhibitor (pembrolizumab) (n?=?11) and HT?+?ACT?+?CT (n?=?12). Peripheral blood was collected to analyze TCR repertoire, measurements of cytokines levels and lymphocyte sub-populations before and after treatment.

Results: The objective response rate (ORR) was 30% (10/33), including three complete responses (CR) (9.1%) and seven partial responses (PR) (21.2%) and a disease control rate (DCR?=?CR?+?PR?+?SD) of 66.7% (22 of 33). The most common adverse reactions, blistering, subcutaneous fat induration, local heat-related pain, vomiting and sinus tachycardia, were observed in association with HT. IL-2, IL-4, TNF-α, and IFN-γ levels in peripheral blood were significantly increased among the clinical responders (p?<?0.05) while IL-6 and IL-10 were elevated among those with progressive disease (p?<?0.05). Peripheral blood CD8⁺/CD28⁺ T cells increased (p?=?0.002), while the CD4⁺/CD25⁺/CD127⁺Treg cells decreased after therapy (p?=?0.012). TCR diversity was substantially increased among the clinical responders.

Conclusions: Combining HT with ACT plus either CT or anti-PD-1 antibody was safe, generated clinical responses in previously treated advanced cancers, and promoted TCR repertoire diversity and favorable changes in serum IL-2, IL-4, TNF-α, and IFN-γ levels in clinical responders.