A study of Hodgkin/Reed-Sternberg cells using single cell polymerase chain reaction

OBJECTIVE: To investigate the characteristics of Hodgkin/Reed-Stemberg (H/R-S) cells found in patients with various types of Hodgkin's disease (HD). METHODS: H/R-S cells were micropicked from frozen sections of tissues affected by HD. The DNA from these cells was amplified by polymerase chain reaction (PCR) using immunoglobulin heavy chain gene FR III a/JH primers and light chain gene family-specific primers. RESULTS: A total of 52/135 (35.8%) isolated cells showed the specific products in the reactions. IgH and V kappa 4 rearrangements were repeatedly found in many cells from a lymphocyte predominance type sample; repeated V kappa 4 and individual IgH/V kappa 2,4 rearrangements and individual IgH, V lambda 3/ V kappa 4 rearrangements were found in two different cases of the nodular sclerosis type; repeated IgH/ V lambda 3 and individual V lambda 2,4 rearrangements, repeated V kappa 2,4 rearrangements, repeated V kappa 4 and individual IgH/ V kappa 3 rearrangements, repeated IgH and individual V kappa 3/ V lambda 4 rearrangements were detected in 3 cases of the mixed cellularity type. Repeated and individual IgH rearrangements were found in other 2 cases. CONCLUSION: The H/R-S cells isolated from the lymphocyte predominance subtypes of HD have IgH and V lambda 4 gene rearrangements. This suggests that the lymphocyte predominance type is a proliferation of neoplastic B cells. The cells isolated from the mixed cellularity and nodular sclerosis types derive from B lineage cells at various stages of differentiation because of the presence of their IgH, kappa and/or lambda gene rearrangements. To our knowledge, this is the first time that the lambda gene rearrangement was detected in H/R-S cells.

Correlation between Increased Circulating Endothelial Progenitor Cells and Stage of non-Hodgkin Lymphoma

This study aims to examine the levels of circulating endothelial progenitor cells （cEPCs） in the peripheral blood of patients with non-Hodgkin lymphoma （NHL） and their correlation with the tumor stage. Forty-one patients with biopsy-proven NHL and 16 healthy individuals were recruited. Pe- ripheral blood mononuclear cells （PBMCs） were isolated by density gradient centrifugation, and cEPCs were characterized by triple staining using antibodies against CD133, CD34 and vascular endothelial growth factor receptor-2 （VEGFR-2, CD309） and quantified by flow cytometry. In NHL patients, the number of cEPCs was significantly greater than in control group （P=-0.000）. The cEPCs counts in patients with NHL of stage III-1V were significantly greater than in stage I -II （P=-0.010）. FACS analysis revealed that the number of cEPCs in NHL patients had no correlation with the gender （P=0.401） or the pathological category （P=0.852）. It was suggested that the over-expression of cEPCs in NHL patients may serve as a novel biomarker for disease progression in NHL.

Composite diffuse large B-cell lymphoma and classical Hodgkin's lymphoma of the stomach:Case report and literature review

The combination of classical Hodgkin’s lymphoma(cHL)and non-Hodgkin lymphoma coexisting in the same patient is not common,especially in one extranodal location.Here we present a rare case of composite diffuse large B-cell lymphoma(DLBCL)and cHL occurring simultaneously in the stomach of a 53-year-old female who presented with upper abdominal discomfort and gas pain.Surgery was performed and the disease was diagnosed pathologically as composite lymphoma of DLBCL and cHL using hematoxylin-eosin and immunohistochemical staining.Epstein-Barr virus(EBV)infection was not detected by in situ hybridization for EBV-encoded RNA or immunohistochemistry for EBV latent membrane protein-1.Polymerase chain reaction analysis from the two distinct components of the tumor demonstrated clonal immunoglobulinκlight chain gene rearrangements.The patient died approximately 11 mo after diagnosis in spite of receiving eight courses of the CHOP and two courses of the rituximab-CHOP(RCHOP) chemotherapy regimen.This case report showed that the two distinct components,DLBCL and cHL,appeared to originate from the same clonal progenitor cell,and that EBV infection was not essential for transformation during the course of tumorigenesis.

Autologous peripheral blood stem cell mobilization following dose-adjusted cyclophosphamide, doxorubicin, vincristine and prednisolone chemotherapy alone or in combination with rituximab in treating high-risk non-Hodgkin's lymphoma

Background: The regimen of cyclophosphamide, doxorubicin, vincristine, and prednisolone(CHOP) is an eicient treatment of non-Hodgkin's lymphoma(NHL). This study aimed to assess the eicacy and toxicity of dose-adjusted CHOP alone or in combination with rituximab(R-CHOP) by examining the stem cell mobilization in NHL patients. Factors afecting the collection of CD34+ cells were also explored.Methods: Our retrospective study included 39 patients eligible for autologous stem cell transplantation: 14 patients who expressed CD20 and were inancially eligible received R-CHOP for autologous peripheral blood stem cell(APBSC) mobilization; the remaining 25 patients received CHOP.Results: The median CD34+ cell yield was 7.01 × 106 cells/kg body weight(range 1.49–28.39 × 106 cells/kg body weight), with only two patients failing to meet the target CD34+ cell harvest of ber of apheresis procedures per patient was 1(range 1–3). The≥2.0 APBS× 106 cells/kg body weight. The median numC mobilization yield of the CHOP group appeared to be higher than that of the R-CHOP group(P response(CR) rate in = 0.005), whereas the success rate was similar between groups. R-CHOP elevated the completeB cell lymphoma patients as compared with CHOP(P = 0.01). No signiicant diferences in toxicity or engraftment were observed between the two groups.Conclusion: The present study demonstrated that dose-adjusted CHOP chemotherapy efectively mobilized APBSCs in NHL patients and that the addition of rituximab to dose-adjusted CHOP chemotherapy elevated the CR rate for patients with B-cell lymphoma.

HCV infection, B-cell non-Hodgkin's lymphoma and immunochemotherapy: Evidence and open questions

There is plenty of data confirming that hepatitis C virus (HCV) infection is a predisposing factor for a B-cell non-Hodgkin's lymphoma (B-NHL) outbreak, while relatively few reports have addressed the role of HCV in affecting B-NHL patients' outcome. HCV infection may influence the short-term outcome of B-NHL because of the emergence of severe hepatic toxicity (HT) during immunochemotherapy. Furthermore, the long term outcome of HCV-related liver disease and patients' quality of life will possibly be affected by Rituximab maintenance, multiple-lines of toxicity during chemotherapy and hematopoietic stem cell transplantation. In this review, data dealing with aggressive and low-grade B-NHL were separately analyzed. The few retrospective papers reporting on aggressive B-NHL patients showed that HCV infection is a risk factor for the outbreak of severe HT during treatment. This adverse event not infrequently leads to the reduction of treatment density and intensity. Existing papers report that low-grade B-NHL patients with HCV infection may have a more widespread disease, more frequent relapses or a lower ORR compared to HCV-negative patients. Notwithstanding, there is no statistical evidence that the prognosis of HCV-positive patients is inferior to that of HCV-negative subjects. HCV-positive prospective studies and longer follow-up are necessary to ascertain if HCV-positive B-NHL patients have inferior outcomes and if there are long term sequels of immunochemotherapies on the progression of liver disease.

HIGH DOSE CHEMORADIOTHERAPY WITH AUTOLOGOUS HEMATOPOIETIC STEM CELL TRANSPLANTATION IN THE TREATMENT OF ADVANCED HODGKIN's LYMPHOMA:A REPORT OF 11 CASES

Objective: High dose therapy (HDT) with autologous hematopoietic stem cell transplantation (ASCT) has become one of the important salvage treatments for the Hodgkin抯 Lymphoma patients with relapsed or resistant disease, but its role as the primary treatment remains indefinite. This study was designed to further evaluate its status in the combined modality treatment, especially, to discuss its value in the primary treatment of the patients who had advanced disease with poor prognostic factors. Methods: Eleven patients who had advanced or relapsed disease with poor prognostic factors were enrolled in this study. Among them, 9 cases had primary treatment, and 2 cases had secondary treatment; one patient received autologous bone marrow transplantation (ABMT), and 10 patients received autologous peripheral blood stem cell transplantation (APBSCT). After induction treatment 4 cases achieved complete response (CR) and 7 cases achieved partial response (PR). High dose chemotherapy combined with total body irradiation (TBI) or total lymph node irradiation (TLI)/subtotal lymph node irradiation (STLI) were adopted in 7 cases and only high dose chemotherapy were adopted in 4 cases as the transplant preparative regimens. 5 cases received complementary irradiation in the primary sites after transplant. Results: The patients who had CR before transplantation were given consolidative therapy. Among the rest with PR, 2 cases achieved CR, 1 case PR, and 4 cases SD. Furthermore all these patients who maintained SD had bone involvement. With a median follow-up for all patients of 13(1-80) months, all of them are alive currently. Four cases are event-free survival (EFS); 4 cases with bone involvement are progression-free survival (PFS); 3 cases experienced relapse after transplant, one of them is EFS for 42 months again after a local relapsed site irradiation; the other two cases are being given further salvaged treatment now. According to the Life Tables method, the cumulative probability of 6-year PFS and OS is 55.68% and 100% respectively. The dominating transplant- related toxicity was bone marrow suppression in grades IV. No obvious cardiac, hepatic, and nephritic toxicity was found. No transplant related mortality. Conclusion: HDT combined with ASCT is a method worthwhile to further study for the treatment of the patients with advanced or relapsed Hodgkin抯 Lymphoma with poor prognostic factors.

THE ULTRASTRUCTURAL STUDY OF NON－HODGKIN＇S LYMPHOMA CELL，CHRONIC LYMPHOCYTIC AND HAIRY CELL LEUKEMIA

Non-Hodgkin’s lymphoma cell leukemia (NHLCL),chronic lymphocytic leukemia (CLL) and hairy cell leukemia (HLC) are the diseases very similar to each other. The differential diagnosis is very difficult,especially when there are small lymphoid cells in Periphcral blood and bone marrow under light microscope. We have observed 34 cases with electron microscope. The studies were correlated with clinical manifestation, cytology, pathology and immunologic histochemistry. Ultrastructural features strongly indicated the difference in three various diseases, although all the immunologic markers showed B-cell type.It is concluded that electron microscopic examination is of a definite significance in the diaguosis and successful treatment.

‘Les liaisons dangereuses’: Hepatitis C, Rituximab and B-cell non-Hodgkin’s lymphomas

Rituximab has provided a revolutionary contribution to the treatment of B-cell non-Hodgkin’s lymphomas (NHL). A high prevalence of hepatitis C virus (HCV) infection has been described in B-cell NHL patients. Cases of liver dysfunction in HCV-positive patients have been reported with Rituximab-containing regimens. In this paper we review the recent data regarding the effects of Rituximab in NHL patients with HCV infection. We also added a section devoted to improving communication between oncohaematologists and hepatologists. Furthermore, we propose a common methodological ground to study hepatic toxicity emerging during chemotherapy.

Therapeutic Efficacy of L-asparaginase in the Treatment of Refractory Midfacial Peripheral T-Cell Non-Hodgkin’s Lymphoma

Objective: To improve the efficacy of refractory midfacial peripheral T-cell non-Hodgkin’s lymphoma (MPTC-NHL) with L-asparaginase (L-ASP) based salvage chemotherapy. Methods: 21 patients with refractory MPTC-NHL were analyzed. 11patients (L-ASP group) received L-asparaginase based salvage chemotherapy consisting of L-asparaginase, vincristine and dexame-thosone. 10 patients (control group) received salvage combination chemotherapy without L-asparaginase. Results: Complete remission rates were 45.6% for L-ASP group and 0.0% for control group (p<0.05). Overall response rates (CR+PR) were 63.6% for L-ASP group and 10.0% for control group, respectively (p<0.05). 2-year survival rates were 45.5% for L-ASP group and 0.0% for control group (p<0.05). The major adverse effects of L-ASP were leukopenia, elevation of serum bilirubin and hyperglycemia. Conclusion: The preliminary clinical study shows that the L-ASP based salvage chemotherapy may improve the response rate and 2-year survival rate of the patients with refractory MPTC-NHL. It is necessary to continue the study further.

淋巴结滤泡辅助T细胞淋巴瘤伴霍奇金/Reed-Sternberg样细胞临床病理学特征、鉴别诊断及基因遗传学特征

目的探讨淋巴结滤泡辅助T细胞淋巴瘤伴霍奇金/Reed-Sternberg(HRS)样细胞的临床病理学特征、鉴别诊断及基因遗传学特征。方法对5例淋巴结滤泡辅助T细胞淋巴瘤(nTFHL)伴HRS样细胞的临床资料、淋巴结活检组织形态学、免疫组化(IHC)、T/B细胞克隆性评估及NGS(二代测序)特征进行分析。结果5例患者,男性4例,年龄分别为65、40、69、75岁;女性1例,年龄57岁。IHC结果示:异型淋巴细胞表达CD3、CD5、PD-1、BCL-6、CD4、CD10,Ki-67阳性率40~70%;HRS样细胞表达CD30、CD20、PAX-5、C-MYC、MUM-1阳性,Ki-67阳性率90%;1例HRS样细胞表达BOB-1,不表达OCT-2;3例HRS样细胞表达CD15、1例HRS样细胞不表达CD15、1例未行CD15染色;EBV原位杂交示:5例HRS样细胞均为阳性。T/B细胞克隆性评估结果:5例TCRG基因检测到单克隆重排,伴或不伴有IGH基因重排。NGS检测结果示:5例均检测到RHOA基因G17V及TET2基因突变。综合淋巴结形态学、IHC、基因重排及NGS结果诊断为nTFHL伴HRS样细胞。结论nTFHL罕见病例可出现HRS样细胞,这些细胞呈现B细胞免疫表型。肿瘤性T细胞经TCR重排研究证实为克隆性。此外,在肿瘤性T细胞中还发现了RHOA和TET2突变。根据文献报道,TET2突变为导致nTFHL发展的第二次打击RHOA突变奠定了基础。若要深入探讨HRS细胞对于nTFHL细胞遗传学的影响,显微切割HRS样细胞进行单细胞测序及基因检测可能更能说明问题。

Allogeneic and autologous stem cell transplantation with busulfan, cyclophosphamide, and etoposide conditioning therapy for relapsed/refractory non-Hodgkin lymphoma

The optimal stem cell transplantation (SCT) conditioning therapy for relapsed/refractory non-Hodgkin lymphoma (NHL) is not clearly defined. In a retrospective analysis, we examined 25 patients with “high risk” relapsed/refractory NHL who received busulfan, cyclophosphamide, and etoposide (Bu/Cy/VP16) conditioning with autologous or allogeneic SCT. The majority of patients had aggressive histology and 52% had primary refractory NHL. Furthermore, 48% of patients had chemotherapy-resistant disease at the time of SCT. Fifty-six percent of patients underwent allogeneic SCT, while 44% had autologous SCT. The median engraftment time for neutrophils and platelets was 13.5 and 14 days, respectively. The 100-day treatment-related mortality (TRM) was 16%, while the 2-year non-relapse mortality (NRM) rate was also 16%. At a median follow-up of 15 months, the estimated 2-year disease-free survival (DFS) rate was 64% (95% confidence interval (CI): 36%-82%) and the estimated 2-year overall survival (OS) was 69% (95% CI: 40%-86%). Furthermore, the 2-year disease-specific survival (DSS) rate was 73% (95% CI: 40%-90%). Using Cox proportional hazard modeling, the International Prognostic Index at time of relapse predicted DFS and OS. Altogether, Bu/Cy/VP16 was associated with early TRM;however, late toxicities (including NRM) were uncommon resulting in relatively good survival rates in a high-risk relapsed/refractory NHL population.

Unusual Presentation of Non-Hodgkin’s B-Cell Lymphoma with Unilateral Right Limb Lymphedema

Background: In clinical practice and setting of general practice it is common to see patients with leg edema. To correctly identify the etiology of the edema and then properly manage the cause is not always easy. The unilateral lymphedema of the lower limb has rarely been reported as an initial presentation for lymphoma, especially in females, usually without classic signs or symptoms, but often with inguinal lymphadenopathy or abdominal masses. Case Report: In this article, we report a rare case of unilateral lower limb edema in a healthy obese woman who complained about the appearance of the disease for several months and for whom deep vein thrombosis and other diseases had been excluded. The histological examination of the biopsy of an enlarged lymph node in the right groin, which was compressing the iliac and femoral vein, revealed the presence of B cell non-Hodgkin lymphoma with high-grade malignancy. Conclusions: A common challenge for primary care physicians is to determine the cause and find an effective treatment for leg edema of unclear etiology. Non-Hodgkin’s B-cell Lymphoma should be considered in the differential diagnosis in patients with unilateral leg edema when the swelling is chronic and deep venous thrombosis is promptly excluded.

非霍奇金淋巴瘤患者自体外周血造血干细胞动员采集的影响因素分析

目的:分析非霍奇金淋巴瘤患者自体外周血造血干细胞动员采集的影响因素。方法:收集西安交通大学第一附属医院血液内科2011年1月至2020年6月共计85例行自体外周血造血干细胞动员采集的非霍奇金淋巴瘤患者临床资料。采用单因素和多因素分析性别、年龄、疾病分型、Ann Arbor分期、移植前状态、国际预后指数评分、动员前化疗次数及动员方案对造血干细胞采集效果的影响。结果:单因素分析发现患者年龄、疾病分型、分期、动员前状态与动员采集CD34^(+)细胞数、动员采集成功率无相关性(P>0.05);男性患者CD34^(+)细胞数多于女性(P=0.01);动员前化疗周期≤4次是采集的有利因素(P<0.05);CHOP-E及CHOP-MTX联合G-CSF动员方案均可作为自体外周血造血干细胞有效的动员方案。多因素结果显示采集前化疗周期数是影响造血干细胞采集成功的独立因素,动员前化疗周期>4次是造血干细胞能否采集成功的不良因素。结论:CHOP-MTX+G-CSF动员方案比较CHOP-E+G-CSF采集CD34^(+)细胞数更多。男性患者、动员采集前化疗周期数较少时更有利于NHL患者自体外周血造血干细胞动员采集。

Non Hodgkin’s Lymphoma with Right Atrial Intra Cardiac Metastases

Background: Diffuse Large B-Cell Lymphoma (DLBCL) is the most variant of Non-Hodgkin’s Lymphoma (NHL) and also the most common variant with secondary intracardiac masses. Case summary: 7 years old child presented to emergency with acute decompensated cardiac failure, ascites and tender hepatomegaly. 2D echo evaluation was suggestive of large intracardiac mass in the right atrium almost completely obstructing Tricuspid valve orifice, gross pericardial effusion and dilated Inferior Vena Cava (IVC). Emergency tumor excision surgery was performed which revealed 4 × 4 cm pinkish firm mass arising from anterior Tricuspid annulus which was completely excised. Child was extubated on postoperative day (POD) 0 and was on minimal inotropic support. Ascites reduced significantly on POD1 allowing abdominal palpation which revealed a mass in the epigastric region. This prompted evaluation by pediatrician and oncology workup suggestive of increased 18-Flouro Deoxy Glucose (18-FDG) uptake in the mediastinum, abdomen, bilateral proximal thighs, all mediastinal lymph nodal stations, bilateral lung hilar stations 10R, 10L involving all encasing the heart and great vessels with pleural deposits, Celiac trunk, superior Mesenteric Artery (SMA), Portal vein, IVC and abdominal aorta. Histo pathology Examination (HPE) and Immuno Histo Chemistry (IHC) of intracardiac mass revealed DLBCL which is metastatic in nature. Chemotherapy was started as per (French American British Lymphomes Malins B) FAB LMB-96 protocol with the child currently in the Induction phase having poor prognosis and less survival interval. Conclusion: Surgery can be considered a treatment option for metastatic intracardiac masses during emergency scenarios like cardiogenic shock to relieve obstruction along the pathway of blood flow in the heart even though we may not be able to completely excise the tumor surgically.

淋巴瘤患者行干细胞移植联合CD30嵌合抗原受体T细胞输注治疗的护理

目的 总结6例复发/难治性CD30阳性淋巴瘤患者进行自体干细胞移植联合抗CD30嵌合抗原受体T细胞输注治疗的护理经验。方法 对6例复发/难治性CD30阳性淋巴瘤患者,完善治疗前的护理评估,重点落实预处理毒性、细胞因子释放综合征以及植入综合征的护理,并将心理支持及健康教育纳入整个治疗过程。结果 6例患者均成功植入造血干细胞,外周血中检测到持续性CAR30转基因,其中5例完全缓解,1例部分缓解。5例出现细胞因子释放综合征,均为Ⅰ级,未观察到神经毒性。随访20.4(12.1,34.4)个月,患者均存活并保持其反应性。结论 自体干细胞移植联合抗CD30嵌合抗原受体T细胞输注治疗具有良好的耐受性和高度活性,护理在治疗各阶段以及毒副反应管理中发挥重要作用。

经典型霍奇金淋巴瘤中CXCR5的表达及临床病理意义

目的探讨经典型霍奇金淋巴瘤(classic Hodgkin lymphoma,CHL)中CXCR5的表达及意义。方法采用免疫组化EnVision两步法检测33例CHL中CXCR5的表达,分析CXCR5在CHL四种亚型中的表达情况及临床病理诊断意义;同时收集10例ALK阳性间变性大细胞淋巴瘤(anaplastic large cell lymphoma,ALCL)和10例ALK阴性ALCL作为对照组,对比分析CXCR5的表达情况。结果33例CHL中,31例CXCR5阳性(93.94%):其中结节硬化型15例(15/16,93.75%)、混合细胞型12例(12/13,92.31%)、淋巴细胞丰富型2例,淋巴细胞消减型2例。CHL中CXCR5的表达情况分别为:33例CD30阳性和PAX5弱阳性中31例阳性(93.94%);14例CD15阴性中12例阳性(85.71%);26例CD20阴性中24例阳性(92.31%);6例LMP1阴性中5例阳性;11例EBER阴性中10例阳性(90.91%)。对照组20例ALCL中,肿瘤细胞CXCR5均阴性。结论CHL中CXCR5阳性率较高,当肿瘤细胞中CD15、LMP1和CD20阴性,或EBER阴性时,CXCR5仍有较高的阳性率,有助于CHL的诊断。尤其是组织形态学和免疫表型不典型的CHL病例,可借助免疫组化标记CXCR5与ALCL鉴别。

PD-1在外周血Treg细胞中的表达与不同病理、分期、治疗方式非霍奇金淋巴瘤患者预后相关性分析

目的 探讨PD-1在外周血Treg细胞中的表达与不同病理、分期、治疗方式非霍奇金淋巴瘤患者预后的相关性。方法 选择2020年1月至2021年4月于我院接受治疗的非霍奇金淋巴瘤(NHL)患者共计74例,所有患者均经病理学组织检查确诊。统计所有患者淋巴瘤病理类型、临床分期以及治疗方式,测定患者程序性细胞死亡受体(PD-1)在患者外周血调节性T细胞(Treg)中的表达水平,并根据PD-1测定结果进行COX回归,分析患者预后情况。结果 (1)本研究74例患者分为:15例前驱淋巴性肿瘤(20.27%),21例惰性NHL(28.38%),19例侵袭性NHL(25.68%),19例成熟T细胞和NK细胞淋巴瘤(25.68%)。临床分期:Ⅰ期4例(5.41%),Ⅱ期23例(31.08%),Ⅲ期36例(48.65%),Ⅳ期11例(14.86%)。治疗方式:放疗16例(21.62%),联合化疗29例(39.19%),靶向药物治疗11例(14.86%),手术切除18例(24.32%)。(2)将侵袭性NHL作为对比因素,惰性NHL(P=0.000,HR=0.205)、前驱淋巴性肿瘤(P=0.013,HR=0.331)、成熟T细胞和NK细胞淋巴瘤(P=0.037,HR=0.329)的患者生存情况更好;将临床Ⅳ期作为对比因素,临床Ⅱ期(P=0.013,HR=0.252)、临床Ⅰ期(P=0.007,HR=0.104)的患者生存情况更好;将手术切除作为对比因素,靶向药物治疗(P=0.031,HR=1.279)的患者生存情况与手术切除类似。(3)测定患者外周血Treg细胞中的PD-1水平,结果显示:在患者不同病理分类、临床分期、治疗方式的分类下,PD-1水平均表现出了差异性(P <0.05),其中,侵袭性NHL、Ⅳ期以及联合化疗的情况下患者PD-1水平最高。结论 PD-1在不同病理类型、临床分期、治疗方式的NHL患者中表达存在差异;PD-1高水平表达的NHL患者预后较差。

端粒酶基因和Ki-67蛋白在Hodgkin淋巴瘤肿瘤细胞中的表达

目的 :研究Hodgkin淋巴瘤R S细胞端粒酶基因表达及与Ki 6 7表达的相关性 ,探讨Hodgkin淋巴瘤的生物学行为。方法 :用原位杂交的方法在 4例正常淋巴结、7例反应性增生淋巴结和 35例Hodgkin淋巴瘤中检测端粒酶基因hTR和hTRT的表达 ,同时应用免疫组化技术检测了Ki 6 7蛋白表达。结果 :端粒酶基因hTRT、hTR在正常淋巴结和反应性增生淋巴结内有低水平或中等量的表达 ,hTRT和hTR分别在 30 / 35、32 / 35例Hodgkin淋巴瘤的R S细胞中具有高表达。Hodgkin淋巴瘤内反应性淋巴细胞hTRT的表达弱 ,且较少 ,而hTR较为广泛。Ki 6 7在R S细胞中的表达与hTRT、hTR较为一致 ,具有明显表达。结论 :Hodgkin淋巴瘤的肿瘤细胞大部分具有hTRT、hTR和Ki 6 7的表达 ,因此R S细胞具有恶性增殖能力。

基于多链结构的CD30 CAR-T细胞的抗肿瘤作用研究

目的基于衔接蛋白DAP12的多链嵌合抗原受体T细胞(CAR-T)开发靶向CD30的CAR-T细胞药物,研究CD30 CAR-T对霍奇金淋巴瘤肿瘤细胞的体外和体内临床前药效。方法通过基因合成和分子克隆技术,设计构建靶向CD30的CAR质粒,进行慢病毒包装,将得到的慢病毒转染T细胞,其中靶向CD30的多链CAR-T为CD30-KIRS2/Dap12-BB组,单链二代CAR-T为CD30-41BBζ组,未做病毒侵染的T细胞为NTD组,利用流式细胞术检测CAR阳性率情况,通过乳酸脱氢酶(LDH)释放检测细胞的杀伤活性,采用酶联免疫吸附试验(ELISA)检测细胞因子干扰素γ(IFN-γ)的分泌水平,进一步通过小鼠异种移植瘤模型检测CD30 CAR-T在小鼠体内抗肿瘤活性。结果靶向CD30的多链CAR-T和单链二代CAR-T进行对比,研究发现多链CAR-T与单链CAR-T的杀瘤作用相似。但值得注意的是,多链CAR-T的IFN-γ分泌水平更高(P<0.001)。更重要的是,在小鼠的肿瘤模型实验中,多链CAR-T实现了肿瘤的完全消退。结论靶向CD30的多链CAR-T在抗肿瘤活性方面优于传统单链CAR-T。

应用多重标记法探讨经典型Hodgkin淋巴瘤H/RS细胞的细胞属性

目的 探讨经典型Hodgkin淋巴瘤H/RS细胞的细胞属性。方法 应用免疫组化多重标记法和原位杂交技术检测CD3、CD20、CD15、CD30、CD10、MPO、bcl 6、κmRNA和λmRNA在 62 例经典型Hodgkin淋巴瘤H/RS细胞的表达。结果 62 例的H/RS细胞CD30 全部阳性,41 例表达 CD15,只有5例表达CD20,均不表达 CD3、bcl 6、CD10、κmRNA、λmRNA。结论 经典型 Hodgkin淋巴瘤的 H/RS细胞为生发中心后期分化的B细胞起源;H/RS细胞对B系标记有较大的变异。它不表达免疫球蛋白轻链κ和λ,可能是这些肿瘤细胞无免疫球蛋白的合成。