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鼻NK/T细胞淋巴瘤瘤细胞属性及其亚型探讨
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摘要
鼻NK(Nature killer)/T细胞淋巴瘤是一类主要发生于淋巴结外的特殊的非霍奇金氏恶性淋巴瘤。鼻腔是最常见的原发部位,因而“鼻NK/T细胞淋巴瘤”可以作为发生在鼻部病例的同义词。该肿瘤与EB病毒感染高度相关,高发于亚洲、墨西哥及中、南美洲,而在欧洲、北美少见。旧称中线恶性网状细胞增生症、多形性网状细胞增生症、致死性中线肉芽肿、恶性肉芽肿、坏死性肉芽肿、血管中心性T细胞淋巴瘤、中线T细胞淋巴瘤及中线外周T细胞淋巴瘤等。该肿瘤侵袭性强,临床常表现为鼻腔及面中线部进行性破坏性改变。组织学形态以突出的凝固性坏死、炎性背景、多形性瘤细胞呈血管中心性生长、浸润并破坏血管壁为特征。长期以来,鼻NK/T细胞淋巴瘤瘤细胞属性问题一直是研究的热点。但由于其在西方发病率低,缺乏大样本深入的研究,对其瘤细胞来源的认识及肿瘤命名方面一直存在争议。
     从目前的研究结果来看,大多数病例似乎是NK细胞肿瘤(EBV+,CD56+),但少数病例具有EBV+,CD56-的细胞毒性T细胞表型,因而推测其肿瘤细胞可能来源于自然杀伤细胞(NK细胞)或毒性T细胞(Tc细胞),并命名其为鼻NK/T细胞淋巴瘤。结合其好发部位、特殊的临床表现,肿瘤细胞的分子学特征(免疫表型和基因型)以及其与EB病毒的密切关系,在新的2001年WHO淋巴造血系统肿瘤分类中,鼻NK/T细胞淋巴瘤作为一种新的单独病变实体被提出。
     毒性T细胞(Tc细胞或CTL),与NK细胞都属于细胞毒性淋巴细胞,有部分Tc表达NK相关抗原CD56,称为NK样T细胞(NKT)。这样,细胞毒性淋巴细胞就包括Tc、NKT和NK三类。
     现代免疫学研究表明T细胞与NK细胞发育分化关系十分密切,两者具有共同的前体细胞。在不同的生物学调节机制下,NK/T前体细胞选择性分化为T或NK细胞的前体细胞。成熟T细胞按抗原识别受体及生
    
    第四军医大学硕士学位论文
    物学功能方式的不同分为4个亚型,CD4+T辅助细胞(Thl汀h2)和CDS十
    毒性T细胞(Tcl瓜2),CD4+Thl型细胞主要分泌IFN一Y、IL一2和TNF一p
    等细胞因子,介导细胞免疫应答,而CD4+ThZ型细胞主要分泌IL一4、IL一5
    和IL一10等细胞因子,促进抗体的产生,介导体液免疫反应。Tcl和TcZ
    细胞也分别分泌Thl和ThZ样细胞因子。近几年来,有研究表明NK细
    胞也可以分为NKI(分泌IFN一种和NKZ(分泌IL一5和IFN一帕亚型。
     综上所述,虽然WHO最新淋巴瘤分类已将鼻NK八,细胞淋巴瘤单
    独列为一种类型,但仍未解决其亚型问题。鼻部恶性淋巴瘤是否都是
    NKfl,细胞淋巴瘤?有无其它类型淋巴瘤?在鼻NKll,细胞淋巴瘤中,
    NK细胞淋巴瘤的构成比究竟有多大?T细胞淋巴瘤比例如何?T细胞琳
    巴瘤的亚型如何?NK细胞淋巴瘤亚型如何?回答这些问题不仅对鼻咽
    恶性淋巴瘤,特别是WHO分类中作为一种新认识的独立淋巴瘤类型的
    鼻NKfl,细胞淋巴瘤的正确诊断具有十分重要的实际意义,而且对理解
    患者临床表现,鼻NKfl,细胞淋巴瘤病理特征等也具有十分重要的指导
    意义,并且关于鼻NK(NKI,NKZ)汀(Tcl,Tc2)细胞淋巴瘤的亚型
    属性研究尚未见文献报道。
     本研究通过对以往明确诊断为中线恶网、恶性肉芽肿、中线外周T
    细胞淋巴瘤及鼻NKJI,细胞淋巴瘤的41例原发于鼻部区域的NHL进行临
    床病理、免疫表型和抗原受体分子基因重排、EBV感染情况等研究,根
    据2001年WHO标准采用多项指标诊断鼻NK厂1,细胞淋巴瘤,并探讨这
    些指标在诊断中的意义,旨在加深对鼻NK几,细胞淋巴瘤的认识。在此基
    础上,进一步研究其肿瘤细胞属性,初步探讨其瘤细胞亚型情况。
     方法
     收集临床资料:采用WHO分类法进行组织学分型;免疫组织化学
    (IHC)采用SP法,选用的抗体有T细胞标记(CD45RO、CD3。、CD4、
    CDS)、B细胞标记(CD20、CD79a)、NK细胞相关抗原(CD56)、毒性
    颗粒相关蛋白(T细胞内抗原,TIA.l)、EBV基因表达产物(LMP一l)、
    分泌Thl样细胞因子淋巴细胞标记(CXCR3、CD134/0X40)、分泌ThZ
    样细胞因子淋巴细胞标记(CCR4、CD30)等:运用EBERI/2.RNA原位
    杂交(IsH)检测肿瘤组织EB病毒潜伏感染情况;运用聚合酶链式反应
    
    第四军医大学硕士学位论文
    (PCR)检测免疫球蛋白重链基因IgH及T细胞抗原受体p、丫链(TCRp
    及TC助)基因重排情况.
     结果
    1.组织学特征:镜下组织学形态多表现为大片凝固性坏死、炎性背景、
     多形性瘤细胞呈血管中心性生长、浸润并破坏血管壁。根据高倍镜下
     瘤细胞大小可分为多形性小细胞为主型(8/41例,19.5%)、多形性中
     等大小细胞为主型(巧/41例,36.6%)和大小细胞混合型三个亚型
     (18/41例,43.9%)。其中,绝大多数为多形性中等大小细胞为主型
     和大小细胞混合型。
    2.根据免疫组化、原位杂交结合基因重排结果对鼻部NHL分型:①免
     疫组化结果:CD45RO阳性29/4l例(70.7%),CD3£阳性26/4l例
     (63.4%),CDZO阳性5/4l例(12.2%),CD79a阳性6/4l例(14.6%),
     CD56阳性17/4l例(4 1 .5%),TIA一l阳性25/41例(60.9%),LMP一l
     阳性6例(6/41例,14.6%)。②原位杂交结果:EBERI/2 .RNA原位
     杂交(ISH)阳性31/41例(75.6%)。根据最新WHO关于鼻NK月,
     细胞淋巴瘤诊断标准CD3。+、CD56+/-、T
Nasal NK/T-cell lymphoma is a peculiar extranodal lymphoma among non-Hodgkin lymphoma. The tumor is typically located in the nasal cavity and therefore the name "nasal NK/T-cell lymphoma" may be used as a synonym for those cases presenting in the nasal region. It has a close relationship with IFNection of EBV and more prevalent in Asia, Mexico, and Central and South America whereas it is rare in Europe and North America. Nasal NK/T-cell lymphoma was named as malignant midline reticulosis, polymorphic reticulosis, lethal midline granuloma, malignant granuloma, necrosis granuloma, angiocentric T-cell lymphoma, T-cell lymphoma of midline, peripheral T-cell lymphoma of midline etc in the past. The neoplasm is characterized by its aggressive clinical features representing nasal cavity and midfacial destroy and the histological features presenting as mass of coagulative necrosis, a heavy admixture of inflammatory cells, polymorphic medium cell show an angiocentric and angiodestructive growth pattern. The lineage o
    f the neoplasm has long been studied during the past years, but Owning to its low incidence of the disease, the research is far below enough. Its neoplastic cell lineage and the name of the tumor have long been controversial.
    In the present study, most cases appear to be NK-cell neoplasms (EBV+ CD56+) while rare cases show an EBV+ CD56- cytotoxic T-cell phenotype. As a result, it is postulated that the neoplastic cell originated from NK cell (NK) or cytotoxic T-cell (Tc) and it is renamed as nasal NK/T cell lymphoma. Recently, new well-defined disease entities have been described in the WHO lymphoma classification based on tumor cell biology combined with
    
    
    
    anatomical site, clinical features, Epstein-Barr virus (EBV) status, and cell lineage as determined by immunophenotype and genotype.
    Cytotoxic T-cell (Tc or CTL), along with NIC-cell make up of the cytotoxic lymphocyte. Parts of Tc were named as NK like T-cell because they can express NFC-cell associated marker (NKT). So the cytotoxic lymphocytes include in fact Tc, NKT and NK. That is to say the nasal NK/T cell lymphoma maybe originated from either of them.
    Recently immunological research indicates that T cell and NK cell are close related in the growth and differentiation procession, they have common precursor cells. Under different biology medulation mechanism, the precursor NK/T cell differentiated into T or NK cell. Mature T cells exist in four populations, defined by their difference antigen recognized receptor and function. They were CD4+ T helper cell (Thl/Th2) and CD8+ cytotoxic (Tcl/Tc2) subsets. Thl-type CD4 cells primarily secrete cytokines such as IL-2 ,TNF-P and IFN- Y , which mediate cellar immunoreactions whereas Th2 cells secrete cytokines such as IL-4, IL-5, and IL-10, which promote the production of antibody and mediate humoral immunoreactions. Recently, NK cells were also shown to be divisible into NK1 (IFN-r producing) and NK2(IL-5 and IFN-r producing).
    To conclude, the question about the subsets of the neoplastic cell of the nasal NK/T cell lymphoma still hasn't been answered although it has been described as a new well-defined disease entity in the WHO lymphoma classification. Are lymphomas located in the nasal all originated NK/T cell? Are there any other types of lymphoma? How about the proportion of NK and T-cell originated lymphoma in nasal NK/T cell lymphoma? How about the subsets in NK or T-cell lymphoma? To answer these questions will benefit not only the diagnoses of nasal NK/T cell lymphoma or other types of lymphoma but also the understanding of its peculiar clinical and histopathology character. The research of the subsets about nasal NK/T cell lymphoma has't found reported.
    To further understand the importance of the standards for nasal NK/T cell lymphoma as a newly defined entity, 41 cases of lymphoma originated from nasal region that diagnosed as midline reticulosis, malignant granuloma,
    
    
    peripheral T-cell lymphoma of midline and nasal NK/T cell lymphoma in the past were studied for their clinicopathol
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