Recent Advances in Diagnosis and Therapy of Angioimmunoblastic T Cell Lymphoma

Angioimmunoblastic T cell lymphoma (AITL) is a common subtype of mature peripheral T cell lymphoma (PTCL). As per the 2016 World Health Organization classification, AITL is now considered as a subtype of nodal T cell lymphoma with follicular helper T cells. The diagnosis is challenging and requires a constellation of clinical, laboratory and histopathological findings. Significant progress in the molecular pathophysiology of AITL has been achieved in the past two decades. Characteristic genomic features have been recognized that could provide a potential platform for better diagnosis and future prognostic models. Frontline therapy for AITL was mainly depending on chemotherapy and the management of relapsed or refractory AITL is still unsatisfactory with a very poor prognosis. Upfront transplantation offers better survival. Novel agents have been introduced recently with promising outcomes. Several clinical trials of combinations using novel agents are underway. Herein, we briefly review recent advances in AITL diagnosis and the evolving treatment landscape.     

Human Vγ2Vδ2 T cells limit breast cancer growth by modulating cell survival‐, apoptosis‐related molecules and microenvironment in tumors

Innate immune system has been known to play an important role in inhibiting the malignant transformation, tumor progression and invasion. However, the mechanistic basis remains ambiguous. Despite polyclonality of human γδ T cells, Vγ2Vδ2 T cell subset was shown to recognize and limit the growth of various tumors at various degrees. The differential recognition of the tumor cells by Vγ2Vδ2 T cells are yet to be defined. Our study reveals that γδ T cells limit in vitro growth of most breast tumor cells, such as SkBr7 (HER2+), MCF7 (ER+) and MDA‐MB‐231 (ER?) by inhibiting their survival and inducing apoptosis, except BrCa‐MZ01 (PR+) cells. To investigate detail mechanisms of antineoplastic effects, we found that cell death was associated with the surface expression levels of MICA/B and ICAM1. Molecular signaling analysis demonstrated that inhibition of cell growth by γδ T cells was associated with the lower expression levels of cell survival‐related molecules such as AKT, ERK and concomitant upregulation of apoptosis‐related molecules, such as PARP, cleaved caspase 3 and tumor suppressor genes PTEN and P53. However, opposite molecular signaling was observed in the resistant cell line after coculture with γδ T cells. In vivo, antineoplastic effects of γδ T cells were also documented, where tumor growth was inhibited due to the downregulation of survival signals, strong induction of apoptotic molecules, disruption of microvasculature and increased infiltration of tumor associated macrophages. These findings reveal that a complex molecular signaling is involved in γδ T cell‐mediated antineoplastic effects.     

MUC1-Tn-targeting chimeric antigen receptor-modified Vγ9Vδ2 T cells with enhanced antigen-specific anti-tumor activity

Chimeric antigen receptor (CAR) αβ T cell adoptive immunotherapy has shown great promise for improving cancer treatment. However, there are several hurdles to overcome for the wide clinical application of CAR-αβ T cells therapy, including side effects and a limited T cells source from cancer patients. Therefore, we sought to identify an alternative T cell subset that could avoid these limitations and improve the effectiveness of CAR-T immunotherapy. γδ T cells are a minor subset of T cells, which share the characteristic of innate immune cells and adaptive immune cells. Vγ9Vδ2 T cells are a predominant γδ T subset in the circulating peripheral blood. In this study, we investigated the antigen-specific antitumor activity of CAR-Vγ9Vδ2 T cells targeting MUC1-Tn antigen. Vγ9Vδ2 T cells were expanded from peripheral blood mononuclear cells of healthy volunteers with zoledronic acid and interleukin-2. CAR-Vγ9Vδ2 T cells were generated by transfection of lentivirus encoding MUC1-Tn CAR. Cytotoxicity assays with various cancer cell lines revealed that CAR-Vγ9Vδ2 T cells could effectively lyse tumor cells in an antigen-specific manner, with similar or stronger effects than CAR-αβ T cells. However, CAR-Vγ9Vδ2 T cells had shorter persistence, which could be improved with the addition of IL-2 to maintain the function of CAR-Vγ9Vδ2 T cells with consecutive stimulation of tumor cells. Using a xenograft mouse model, we further showed that CAR-Vγ9Vδ2 T cells more effectively suppressed tumor growth in vivo than Vγ9Vδ2 T cells. Therefore, MUC1-Tn CAR-modified Vγ9Vδ2 T cells may represent a novel, promising ready-to-use product for cancer allogeneic immunotherapy.     

Cell Density-Dependent Proliferative Effects of Transforming Growth Factor (TGF)-β1, β2, and β3 in Human Chondrosarcoma Cells HCS-2/8 Are Associated with Changes in the Expression of TGF-β Receptor Type I

In this study, the growth properties of the human chondrosarcoma cell line HCS- 2/8, its response to transforming growth factor (TGF)-β isoforms 1, 2, and 3, and its expression of TGF-β receptors I and II were examined. We demonstrated that these tumor cells are not contact-inhibited and that they can proliferate in the absence of additional serum growth factors. In sparse cultures, all TGF-β forms inhibited the growth of HCS-2/8 cells, whereas they induced a 2-fold increase of DNA synthesis in serum-fed confluent cultures. In serum-free confluent conditions only TGF-β1 stimulated the proliferation rate, whereas TGF-β2 was without effect and TGF-β3 was rather inhibitory. This bimodal effect of TGF-β forms was associated with a greater level of TGF-β receptor I mRNA in confluent HCS-2/8 than in sparse cultures, suggesting that the growth response to TGF-β forms is dependent on the receptor profile expressed.     

NK/T细胞淋巴瘤

目的：了解和认识NK／T细胞淋巴瘤的临床病理和免疫学特点。方法：选用NK细胞特异性抗体CD56，T细胞特异性抗体CD3E，对117例T细胞淋巴瘤进行标记。结果：117例T细胞淋巴瘤中26例CD56（＋），CD3ε（＋），阳性表达率为22．2％，26例中结外淋巴瘤16例（61．5％），其中15例位于鼻部；淋巴结内10例（38．5％），26例中18例（69．2％）表现为细胞的多形性特点，12例（46．2％）有凝固性坏死。结论：NK／T细胞淋巴瘤免疫学特点是CD56与CD3ε共同表达，NK／T细胞淋巴瘤多发生在结外，以发生于鼻部者为多，少数发生在结内，肿瘤细胞的多形性和肿瘤组织的凝固性坏死是主要的病理学特点。     

DNA polymerase β gene mutations in human bladder cancer

We examined 24 human bladder cancer tissues for possible mutations in the entire coding region of the human DNA polymerase β gene using polymerase chain reaction analysis, single-strand conformational polymorphism analysis of RNA, and sequence analysis. DNA polymerase β gene mutations were observed in four of the 24 cases (16.7%) and included three missense point mutations and a single base insertion. The single base insertion was also observed in our previous study of human prostate cancer, suggesting that this region may be a hot spot for mutation of the DNA polymerase β gene. No clinical or pathological association was found among the four cases that contained the mutation. Three of the four cases with DNA polymerase β gene mutation had mutations of the p16 or RB genes or loss of heterozygosity of the p53 and APC gene loci. The results of the study presented here suggest that DNA polymerase β gene mutations, in combination with mutations of tumor suppressor genes, may be involved in certain cases of human bladder cancer. © 1996 Wiley-Liss, Inc.     

The CD226‐ERK1/2‐LAMP1 pathway is an important mechanism for Vγ9Vδ2 T cell cytotoxicity against chemotherapy‐resistant acute myeloid leukemia blasts and leukemia stem cells

Vγ9Vδ2 T cells are attractive effector cells for immunotherapy with potent cytotoxic activity against a variety of malignant cells. However, the effect of Vγ9Vδ2 T cells on chemotherapy‐resistant acute myeloid leukemia (AML) blasts, especially highly refractory leukemia stem cells (LSCs) is still unknown. In this study, we investigated the effect of cytotoxicity of allogeneic Vγ9Vδ2 T cells on chemotherapy‐resistant AML cell lines, as well as on primary AML blasts and LSCs obtained from refractory AML patients. The results indicated that Vγ9Vδ2 T cells can efficiently kill drug‐resistant AML cell lines in vitro and in vivo, and the sensitivity of AML cells to Vγ9Vδ2 T cell–mediated cytotoxicity is not influenced by the sensitivity of AML cells to chemotherapy. We further found that Vγ9Vδ2 T cells exhibited a comparable effect of cytotoxicity against LSCs to primary AML blasts. More importantly, we revealed that the CD226–extracellular signal–regulatory kinase1/2 (ERK1/2)–lysosome‐associated membrane protein 1 (LAMP1) pathway is an important mechanism for Vγ9Vδ2 T cell–induced cytotoxicity against AML cells. First, Vγ9Vδ2 T cells recognized AML cells by receptor‐ligand interaction of CD226–Nectin‐2, which then induced ERK1/2 phosphorylation in Vγ9Vδ2 T cells. Finally, triggering the movement of lytic granules toward AML cells induced cytolysis of AML cells. The expression level of Nectin‐2 may be used as a novel marker to predict the susceptibility/resistance of AML cells to Vγ9Vδ2 T cell treatment.     

IL‐17A‐producing CD30+ Vδ1 T cells drive inflammation‐induced cancer progression

Although it has been suspected that inflammation is associated with increased tumor metastasis, the exact type of immune response required to initiate cancer progression and metastasis remains unknown. In this study, by using an in vivo tumor progression model in which low tumorigenic cancer cells acquire malignant metastatic phenotype after exposure to inflammation, we found that IL‐17A is a critical cue for escalating cancer cell malignancy. We further demonstrated that the length of exposure to an inflammatory microenvironment could be associated with acquiring greater tumorigenicity and that IL‐17A was critical for amplifying such local inflammation, as observed in the production of IL‐1β and neutrophil infiltration following the cross‐talk between cancer and host stromal cells. We further determined that γδT cells expressing Vδ1 semi‐invariant TCR initiate cancer‐promoting inflammation by producing IL‐17A in an MyD88/IL‐23‐dependent manner. Finally, we identified CD30 as a key molecule in the inflammatory function of Vδ1T cells and the blockade of this pathway targeted this cancer immune‐escalation process. Collectively, these results reveal the importance of IL‐17A‐producing CD30⁺ Vδ1T cells in triggering inflammation and orchestrating a microenvironment leading to cancer progression.     

Clinicopathological analysis of 17 primary cutaneous T‐cell lymphoma of the γδ phenotype from Japan

Primary cutaneous γδ T‐cell lymphoma (PCGD‐TCL) is an aggressive lymphoma consisting of clonal proliferation of mature activated γδ T‐cells of a cytotoxic phenotype. Because primary cutaneous γδ T‐cell lymphoma is a rare disease, there are few clinicopathological studies. In addition, T‐cell receptor (TCR) γδ cells are typically immunostained in frozen sections or determined by TCRβ negativity. We retrospectively analyzed 17 primary cutaneous T‐cell lymphomas of the γδ phenotype (CTCL‐γδ) in a clinicopathological and molecular study using paraffin‐embedded sections. Among 17 patients, 11 had CTCL‐γδ without subcutaneous panniculitis‐like T‐cell lymphoma (SPTCL) features and six had CTCL‐γδ with SPTCL features. Immunophenotypically, some significant differences were found in CD8 and CD56 positivity between our patient series of CTCL‐γδ patients with SPTCL features and SPTCL‐γδ patients described in the previous literature. A univariate analysis of 17 CTCL‐γδ patients showed that being more than 60 years old, presence of visceral organ involvement, and small‐to‐medium cell size were poor prognostic factors. In addition, the 5‐year overall survival rate was 42.4% for the CTCL‐γδ patients without SPTCL features and 80.0% for those with SPTCL features. Consequently, there was a strikingly significant difference in overall survival among SPTCL, CTCL‐γδ with SPTCL features and CTCL‐γδ without SPTCL features (P = 0.0005). Our data suggests that an indolent subgroup may exist in CTCL‐γδ. Studies on more cases, including those from other countries, are warranted to delineate the clinicopathological features and the significance in these rare lymphomas.     

Hematopoietic Progenitors and Synergism of Interferon-γ and Stem Cell Factor

Interferon-γ (IFN-γ), an immunoregulatory cytokine produced by activated T cells and natural killer cells in response to viral infection or other stimuli, is generally recognized as a suppressor of hematopoiesis. IFN-γ inhibited in vitro colony formation by granulocyte-macrophage (GM), erythroid and multipotential progenitors. This cytokine exerted direct suppression on the proliferation process, but not on the commitment, of GM progenitors. The antiproliferative effects of IFN-γ may, in part, result from the prolongation of the doubling time of GM progenitors. Clinically, IFN-γ may play an important role in the pathogenesis of pancytopenia in aplastic anemia and in the hemophagocytic syndrome. However, as well as showing inhibitory effects, IFN-γ increased the number of pure and mixed megakaryocyte colonies formed by post-5-fluorouracil treated bone marrow cells and, moreover, the addition of IFN-γ to culture containing stem cell factor resulted in a synergistic effect on the development of both primitive hematopoietic progenitors and mature populations. These findings suggest that IFN-γ has bifunctional activity in hematopoiesis.     

Rap2b基因真核表达载体构建及其对NIH3T3细胞P38通路的影响

目的构建pcDNA3.1-Rap2b真核表达载体,以外源基因Rap2b转染NIH3T3细胞,以了解该基因对NIH3T3细胞AKT、ERK、JNK和P38信号转导通路的影响,为探讨该基因在人肺癌发生中的作用提供实验依据。方法构建人Rap2b真核表达载体pcDNA3.1-Rap2b并稳定转染NIH3T3细胞,利用Western blot方法对转染的细胞进行AKT、ERK、JNK和P38磷酸化蛋白及总蛋白表达水平检测。结果转染pcDNA3.1-Rap2b质粒的细胞与转染空质粒pcDNA3.1的细胞相比,其AKT、ERK、JNK磷酸化蛋白表达水平差异无统计学意义（P＞0.05）,而P38磷酸化蛋白表达水平明显上调（P＜0.05 ）。结论Rap2b基因外源性高表达可能对P38通路有活化作用,对JNK、ERK、AKT通路无活化作用。     

Anti-proliferative activity of heat shock protein (Hsp) 90 inhibitors via β-catenin/TCF7L2 pathway in adult T cell leukemia cells

The aim of this study is to evaluate the effect of heat shock protein 90 (Hsp90) inhibition, and to identify molecular pathways responsible for anti-proliferative effect on adult T cell leukemia/lymphoma (ATL) cells. For Hsp90 inhibition, we used geldanamycin derivates, 17-AAG (17-allylamino-17-demethoxygeldanamycin) and 17-DMAG (17-(dimethylaminoethylamino) 17-demethoxygeldanamycin) in this study. The inhibitory concentration (IC₅₀) of 17-AAG in an ATL cell line, designated as TaY, and two HTLV-1 transformed cell lines (MT-2 and MT-4) was 300–700 nM, and that of 17-DMAG was 150–200 nM. Fresh ATL cells obtained from patients were more sensitive to both 17-AAG and 17-DMAG. Gene expression analysis of TaY cells revealed up-regulation of HSPA1A encoding Hsp70, a hallmark of Hsp90 inhibition. Genes regulating cell proliferation or anti-apoptosis (i.e. BCL2 and BIRC5), genes related to cytokines or chemokines (i.e. IL9 and CCL27), and notably TCF7L2, a down-stream effecter of β-catenin were remarkably down-regulated. Down-regulation of TCF7L2 mRNA was noted in the three cell lines and two patient specimens after Hsp90 inhibition. Hsp90 inhibitors dephosphorylate AKT, thereby, activate GSK-3β, which phosphorylates β-catenin for ubiquitination. This indicates the possibility that β-catenin/TCF7L2 pathway plays an important role in Hsp90 inhibitor-induced cell death in ATL cells and HTLV-1 transformed cells. Our results have provided new insights into the complex molecular pharmacology of Hsp90 inhibitors, and suggest that Hsp90 inhibitors might be beneficial as anti-proliferative agents in treating ATL patients.     

自体干细胞移植治疗T细胞淋巴瘤31例临床分析

目的 探讨自体干细胞移植治疗T细胞淋巴瘤的疗效和其预后因素.方法 回顾性分析31例给予自体干细胞移植治疗的T细胞淋巴瘤的临床资料,观察3年总生存率和无进展生存率,分析一般状况(PS)、乳酸脱氢酶(LDH)、移植前状况、分期、外周T细胞淋巴瘤预后指数(PIT)评分对生存的影响.结果 全组中位随访时间为28(5～68)个月...     

非何杰金氏淋巴瘤的T细胞受体和免疫球蛋白重链基因重排的初步研究

T细胞受体及免疫球蛋白的基因重排研究对鉴定T.B细胞的单克隆增生,协助诊断淋巴瘤有重要意义。本研究用~(32)P标记的Tβ,Tγ,JH探针对26例淋巴瘤及2例急性淋巴细胞白血病标本的DNA进行Southern印迹杂交,发现在11例B细胞淋巴瘤均出现JH基因重排,13例T细胞淋巴瘤中11例出现Tβ链,4例出现Tγ链基因重排,4例出现JH基因重排。2例免疫组化不能确定细胞来源的淋巴瘤、1例有Tγ,另1例有JH重排。2例急性淋巴细胞白血病分别出现Tβ及Tγ重排。该初步结果说明基因重排研究是确定淋巴瘤细胞克隆性增生的有力手段。     

Clinical Significance of the t(14;18) And BCL2 Overexpression in Follicular Large Cell Lymphoma

Follicular large cell lymphoma (FLCL) is an aggressive disease that responds to anthracy-cline-containing chemotherapy much like diffuse large B-cell lymphoma (DLBCL). Since the t(14; 18) and/or bcl2 protein expression are less common in FLCL than in its low-grade counterparts, we sought to determine whether these features were predictive of survival as in DLBCL. We studied 50 patients with FLCL who were treated with curative intent. The t(14; 18) was found by cytogenetic analysis in 56% of the patients and bcl2 protein was expressed by the tumor cells in 73%, but neither was predictive of survival. However, abnormalities of chromosome 17p and the presence of trisomy 21 were adverse predictors of survival, as were a number of clinical features. We conclude that neither the absence of the t(14; 18) nor the lack of bcl2 expression explain the good response of a subset of patients with FLCL to curative therapy.     

Tumor Necrosis Factor-β and Hypercalcemia

Hypercalcemia in hematological malignancy is frequently encountered in lymphoid malignancies such as adult T-cell leukemia (ATL) and multiple myeloma and is difficult to manage. As a causative agent of hypercalcemia in ATL, tumor necrosis factor-β (TNF-β), previously known as lymphotoxin, has been carefully studied and reviewed here. Bone resorption studies showed the presence of activity in culture supernatants of HTLV-I infected cells. Enzyme linked immunosorbent assays (ELISA) for TNF-β detected elevated TNF-β in the sera of ATL patients with hypercalcemia. Immunostaining by monoclonal anti-TNF-β antibody demonstrated the presence of TNF-β in both HTLV-I infected cell lines and freshly isolated ATL cells. Furthermore biological TNF-β activity assay including inhibition of anti-TNF-β antibody confirmed the conventional documentation of TNF-β activity in the sera and culture supernatants of HTLV-I infected cell lines. These studies showed that the TNF-β secreted from ATL cells might be one of the factors contributing to the hypercalcemia in patients with ATL functioning as an osteoclast activating factor (OAF).     

鼻腔鼻窦NK/T细胞淋巴瘤的诊治与预后分析

[目的]探讨鼻腔鼻窦NK/T细胞淋巴瘤的临床特点、治疗方法、预后以及影响预后的相关因素。[方法]回顾分析34例鼻腔鼻窦NK/T细胞淋巴瘤的临床资料,采用KaplanMeier法及Cox回归模型进行生存分析。[结果]鼻腔鼻窦NK/T细胞淋巴瘤总5年生存率、3年生存率及1年生存率分别为5.9%,17.6%,70.6%,中位生存时间为20.0月;Ⅰ期患者1年生存率为100.0%,3年生存率为33.3%;Ⅱ期患者1年生存率为62.5%,3年生存率为31.2%;Ⅲ、Ⅳ期患者1年生存率为66.6%,3年生存率为0。单纯手术治疗者1年生存率为66.6%,3年生存率为16.6%;CHOP方案化疗或术后化疗者1年生存率为50.0%,3年生存率为0;综合放化疗者1年生存率为100.0%,3年生存率为50.0%。有B症状者1年生存率为37.5%,3年生存率为25.0%;而无B症状患者1年生存率为69.1%,3年生存率为19.1%。[结论]鼻腔鼻窦NK/T细胞淋巴瘤预后差,肿瘤的分期、治疗的方式与预后有关,综合放化疗患者疗效明显好于其它治疗方案,放疗是早期NK/T细胞淋巴瘤的主要治疗方法。     

Metabolomics Investigation of Cutaneous T Cell Lymphoma Based on UHPLC-QTOF/MS

Objectives: The identification of cutaneous T cell lymphoma (CTCL) biomarkers may serve as a predictor of disease progression and treatment response. The aim of this study was to map potential biomarkers in CTCL plasma. Design and Methods: Plasma metabolic perturbations between CTCL cases and healthy individuals were investigated using metabolomics and ultrahigh performance liquid chromatography–quadrupole time-of-flight mass spectrometry (UHPLC-QTOF/MS). Results: Principal component analysis (PCA) of the spectra showedclear metabolic changes between the two groups. Thirty six potential biomarkers associated with CTCL were found. Conclusions: Based on PCA, several biomarkers were determined and further identified by LC/MS/MS analysis. All of these could be potential early markers of CTCL. In addition, we established that heparin as a nticoagulant has better pre-treatment results than EDTA with the UHPLC-QTOF/MS appraoch.     

Response to 5‐azacytidine in a patient with TET2‐mutated angioimmunoblastic T‐cell lymphoma and chronic myelomonocytic leukaemia preceded by an EBV‐positive large B‐cell lymphoma

We report the case of a patient with a history of Epstein–Barr virus‐positive large B‐cell lymphoma, who relapsed with an angioimmunoblastic T‐cell lymphoma (AITL) associated with a chronic myelomonocytic leukaemia (CMML). We performed targeted next‐generation sequencing on CMML and AITL DNA, which revealed mutations of TET2, DNMT3A, SRSF2, NRAS and IDH1, thus confirming that the spectrum of AITL mutations share similarities with myeloid disorders. The frequencies of TET2/DNMT3A and SRSF2 variants could support the hypothesis that TET2/DNMT3A mutations occurred in an early progenitor cell, which later progressed to both the AITL and CMML clones. Treatment with 5‐azacytidine led to the complete remission of both diseases. Thus, targeting DNA methylation abnormalities in AITL may be an alternative strategy to chemotherapy. Copyright © 2016 John Wiley & Sons, Ltd.     

两种化疗方案一线治疗外周 T细胞淋巴瘤的临床对照研究

目的：比较EPOCH方案和CHOP方案一线治疗外周T细胞淋巴瘤的疗效及不良反应。方法将2010年1月至2015年1月北京军区总医院收治的32例外周T细胞淋巴瘤患者分为2组。16例采用EPOCH方案化疗：依托泊苷50 mg／m2 d1～4、吡柔比星10 mg／m2 d1～4、长春地辛1 mg／d d1～4、环磷酰胺750 mg／m2 d5、强地松60 mg／m2 d1～5。16例采用CHOP方案化疗：环磷酰胺750 mg／m2 d1、吡柔比星45 mg／m2 d1、长春地辛4 mg／d d1、强的松60 mg／m2 d1～5。每3周左右完成1个化疗周期,共进行6～8个化疗周期,比较2种化疗方案的临床疗效。结果随访至2016年1月,2组患者完全缓解效率分别为56．3％和31．3％,总有效率分别为81．3％和62．5％,中位生存期分别为28．5个月和21．8个月。结论 EPOCH方案一线治疗外周T细胞淋巴瘤较CHOP方案具有显著的优势。