原发性皮肤T细胞淋巴瘤

原发性皮肤T细胞淋巴瘤属结外非霍奇金淋巴瘤 ,以皮肤内辅助T细胞的单克隆扩增为特征。他与有相同组织学亚型的累及皮肤的原发性淋巴结淋巴瘤在临床及组织学特征、生物学行为及预后都明显不同。如今最常用的原发性皮肤T细胞淋巴瘤分类是EORTC分类 ,HTLV -Ⅰ可能与原发性皮肤T细胞淋巴瘤相关 ,有待进一步验证。原发性皮肤T细胞淋巴瘤有较为独特的免疫表型 ,T细胞受体基因重排检测有助于其早期诊断及疾病的转归和预防的判断     

原发性皮肤T细胞淋巴瘤

原发性皮肤T细胞淋巴瘤属结外非霍奇金淋巴瘤，以皮肤内辅助T细胞的单克隆扩增为特征。他与有相同组织学亚型的累及皮肤的原发性淋巴结淋巴瘤在临床及组织学特征、生物学行为及预后都明显不同。如今最常用的原发性皮肤T细胞淋巴瘤分类是EORTC分类，HTLV-Ⅰ可能与原发性皮肤T细胞淋巴瘤相关，有待进一步验证。原发性皮肤T细胞淋巴瘤有较为独特的免疫表型，T细胞受体基因重排检测有助于其早期诊断及疾病的转归和预防的判断。     

皮肤的结外鼻型NK/T细胞淋巴瘤1例

对1例皮肤的结外鼻型NK/T细胞淋巴瘤临床病理结合免疫组化染色、EB病毒原位杂交及T细胞受体基因重排进行分析.右胫后多发结节,组织病理特征为肿瘤组织在真皮及皮下组织内弥漫性浸润,肿瘤组织具有血管中心性及血管破坏性特点,肿瘤细胞具有异型性.瘤细胞表达CD2,CD56,颗粒酶B,EBER阳性,未检测到TCR克隆性基因重排.诊断为皮肤的结外鼻型NK/T细胞淋巴瘤.皮肤的结外鼻型NK/T细胞淋巴瘤恶性度高、预后差;诊断有赖于组织病理及免疫表型检测及EBER原位杂交技术.     

皮肤的结外鼻型NK/T细胞淋巴瘤1例及文献复习

目的报道1例皮肤结外鼻型NK/T细胞淋巴瘤,以引起临床和病理医师对此病的关注。方法通过临床病理分析结合免疫组化染色、EB病毒原位杂交及T细胞受体基因重排的PCR检测分析确诊。结果左膝内后方皮损初次活检诊断为结节性脂膜炎,1个月内再次活检示真皮和皮下脂肪内肿瘤大片坏死,瘤细胞异型性明显,血管中心性浸润和血管坏死。瘤细胞表达CD2,CD8,CD45RO,CD56,TIA-1,GranzymeB和LMP-1,EB病毒(+);未检测到TCR-γ的克隆性基因重排。诊断为皮肤的结外鼻型NK/T细胞淋巴瘤。结论皮肤的结外鼻型NK/T细胞淋巴瘤恶性度高、易误诊、预后差;诊断有赖于常规组织病理结合分子病理技术。     

原发性皮肤间变性大细胞淋巴瘤T细胞γ受体、IgH基因重排

目的：探讨原发性皮肤间变性大细胞淋巴瘤(C-ALCL)临床病理特点和基因诊断方法。方法：对6例C-ALCL的临床表现、病理形态学和免疫组化染色进行观察,并用PCR方法对石蜡标本进行T细胞γ受体(TCRγ)和重链免疫球蛋白(1gH)基因重排检测。结果：临床起病以孤立性结节多见,病情进展缓慢,个别可自行消退。5例患者经治疗病情稳定,1例死于淋巴结及肝脏转移。镜下以75％以上CD30^ 间变性大细胞弥漫浸润真皮及皮下脂肪组织为特点,多数瘤细胞表达T细胞免疫表型。5例标本TCRγ基因重排阳性。结论：C-ALCL是少见的原发皮肤的低度恶性T细胞性淋巴瘤,预后较好。综合临床表现、组织病理改变、免疫组化及基因重排检测有助于本病的正确诊断。     

T细胞受体基因重排检测皮肤T细胞淋巴瘤

为了研究基因诊断在皮肤Ｔ细胞淋巴瘤的临床应用，采用聚合酶链反应技术，对６０例皮肤淋巴细胞浸润疾患进行了Ｔ细胞受体基因重排的检测。结果显示，检出ＴＣＲ－β和ＴＣＲ－γ基因克隆重排的有：３６／４０例ＣＴＣＬ，４／６例可疑蕈样肉芽肿／Ｓｅｚａｒｙ综合征和１／１例淋巴瘤样丘疹病。     

皮肤T细胞淋巴瘤细胞表型和基因型的研究进展

近年来对皮肤T细胞淋巴瘤的研究发现:皮肤T细胞淋巴瘤细胞的某些表型可能在疾病发展过程中丢失,或表达一些奇特的新表型;Southern印迹分析检测出T细胞受体基因重排带,提示该病是一种克隆性恶性疾病。应用多种单克隆抗体检测皮肤T细胞淋巴瘤细胞表型的变化及Southern印迹技术检测克隆性重排带可能对该病的早期诊断,鉴别诊断和预后判定具有意义。     

皮肤假性淋巴瘤石蜡包埋组织克隆性基因重排检测

皮肤假性淋巴瘤系指在组织学上类似皮肤恶性淋巴瘤,而临床表现为良性生物学行为,不完全符合皮肤淋巴瘤的诊断标准.区分淋巴细胞的良性反应性增生和恶性增生(淋巴瘤)一直是临床病理诊断的难题^[1].淋巴细胞成熟过程中免疫球蛋白和T细胞受体基因重排,为检测淋巴细胞的克隆性增生提供了理论基础^[2].     

蕈样肉芽肿T细胞抗原受体γ基因重排的研究

蕈样肉芽肿(MF)是一种低度恶性的皮肤T细胞淋巴瘤。淋巴细胞恶性增殖的主要特点是克隆性增殖,在T淋巴瘤表现为TCR的单克隆性重排,而绝大多数炎症及反应性淋巴细胞浸润则是表现为多克隆性的。根据此点,近十几年来,国外用Southern杂交或聚合酶链反应检测皮损的T淋巴细胞的克隆性,主要是检测TCR-β和γ基因重排。由于γ基因重排可出现在几乎所有的T淋巴细胞及T淋巴细胞瘤,且其结构相对简单,故与Southern印迹方法相比用聚合酶链反应(PCR)方法检测TCR-γ基因重排更简便易行,且检出阳性率较高。     

原发性皮肤T细胞淋巴瘤T细胞受体基因重排检测

Southern印迹分析（SBA）和取和合酶链反应（PCR）法检测早期（IA）和组织学上未确诊的蕈样肉芽肿（MF）皮肤损害的T细胞受体基因重排（TCRGR）阳性率分别为50％以上和19％，有助于诊断。IIA期MF患者特别是伴浅表淋巴结肿大（组织学上大都是反应性增生）的外周血中TCRGR阳性率较高（65％－80％），支持MF早期即为一系统性疾病的理论，但外周血中克隆T细胞和预后相关性则难于确定。Sezary综合征（SS）的TCRGR的发生率在皮肤、淋巴结和外周血中分别为70％、100％和86％。非MF／SS皮肤T细胞淋巴瘤的皮肤 损害和外周血中TCRGR亦较常见。淋巴瘤样丘疹病的皮肤损害和外周血中也可见TCRGR。     

Cellular coincidence of clonal T cell receptor rearrangements and complex clonal chromosomal aberrations-a hallmark of malignancy in cutaneous T cell lymphoma

Detection of a clonal T cell receptor (TCR) gene rearrangement is used in the diagnosis of primary cutaneous T cell lymphomas (CTCL) whereas chromosomal aberrations serve as a diagnostic tool for leukaemias and nodal lymphomas. To what extent both approaches specify the same cell population remains unknown. We investigated the coincidence of TCR clonality with complex clonal chromosomal aberrations, indicating qualitative alteration of the affected cells, in 17 CTCL patients. Out of 41 skin, blood, and lymph node samples studied, 34 gave results in chromosome and TCR analyses. With 88%, most specimens revealed corresponding results by both techniques (27 of 34 clonal, three of 34 non-clonal). In two patients, analysis of micro-dissected cells demonstrated that neoplastic T cells bear both a dominant TCR rearrangement and a complex chromosomal aberration. The cutaneous clone was found in blood samples of 11 of 12 patients (including early stages), and investigation of follow-up skin and blood samples indicated persistence of the T cell clone in 11 of 14 cases. In conclusion, we show that dominant TCR clones and chromosomal clones converge in all stages of CTCL. These clones disseminate into blood and skin at early disease stages and persist despite therapy. The coexistence of a dominant TCR clone and a clonal chromosomal aberration can thus be used as a hallmark of malignancy.     

New insights into the applicability of T-cell receptor gamma gene rearrangement analysis in cutaneous T-cell lymphoma

BACKGROUND: Detection of clonal T-cell receptor (TCR) gamma gene rearrangement by polymerase chain reaction (PCR) based method is a marker for cutaneous T-cell lymphoma (CTCL) although it can be seen in some benign dermatoses. To determine the accuracy of histologic criteria alone as well as the adjuvant diagnostic role of TCR gene rearrangement for the diagnosis of CTCL, we studied 100 patients with cutaneous T-cell infiltrates by both histology and TCR gene rearrangement. METHODS: The histologic features of the 100 patients were first reviewed by two independent dermatopathologists and their confidence in the diagnosis of CTCL was assigned one of four levels. Then the specimens were analyzed for TCR gene rearrangement either on paraffin-embedded or fresh-frozen tissue by PCR/denaturing gradient gel electrophoresis (DGGE). RESULTS: The clonality was detected in 100% (15/15) diagnostic of, 84.6% (11/13) consistent with, 57.6% (19/33) suggestive of CTCL. In 9 cases TCR gene rearrangement was compared between formalin-fixed and fresh specimens of the same individual, but with different degrees of histologic confidence (no lower than suggestive). In all cases fresh specimens were positive. In 5 of the cases (2-diagnostic, 2-consistent, 1-suggestive) formalin-fixed specimens were positive as well, and in 4 cases (1-consistent, 3-suggestive) formalin-fixed specimens were negative. When TCR gene rearrangement was studied in eight cases on sequential biopsies from the same patient, the clonality was detected in only one or two biopsies in four cases in which the histologic confidence was low (suggestive or nondiagnostic). The TCR gene rearrangement study showed identical banding patterns in lesions from different clinical stages in most patients. However, we observed that in one case, oligoclonal-banding pattern was seen in initial biopsy with histopathologic consistent with CTCL, while monoclonal banding pattern in more advanced lesion. CONCLUSIONS: Our data have demonstrated that TCR gene rearrangement studies by PCR/DGGE are consistently positive regardless of tissue fixation (formalin-fixed, paraffin-embedded vs. fresh-frozen tissue) and biopsy site when the histologic degree of confidence is very high (diagnostic). So, it may be of less importance as an adjuvant to histopathologic diagnosis for the cases with diagnostic CTCL histology. However, TCR gene rearrangement studies are particularly important in earlier cases with less conclusive histology, which provides strong confirmatory evidence of an evolving CTCL. In these cases, multiple biopsies may be required to establish the diagnosis and analysis of fresh tissue is suggested to increases the sensitivity. Moreover, our observation also suggested that some CTCL might not be monoclonal de novo, but oligoclonal instead.     

T‐cell receptor γ gene rearrangement in cutaneous T‐cell lymphoma: comparative study of polymerase chain reaction with denaturing gradient gel electrophoresis and GeneScan analysis

Background The usefulness of T‐cell receptor gene rearrangement (TCR‐GR) analyses for differentiating cutaneous T‐cell lymphoma (CTCL) from benign inflammatory disorders (BID) has been insufficiently studied to date. Objectives To evaluate the diagnostic value of TCR‐GR analyses, comparing polymerase chain reaction (PCR) with denaturing gradient gel electrophoresis (DGGE) analysis and BIOMED‐2 standardized protocol PCR with GeneScan analysis (BIOMED‐2‐GS). Methods Both types of PCR were performed in 157 patients evaluated for initial features suggestive of CTCL between 1996 and 2007. After clinical and histological review, the final diagnosis was CTCL in 77 cases and BID in 80 cases. Results DGGE and BIOMED‐2‐GS had a similar diagnostic value for distinguishing CTCL from BID, with a sensitivity of 74% and 77%, respectively, and a specificity of 86%. The observed concordance between both methods was 90% and the kappa coefficient was 0·79. Positivity rates did not depend on the PCR method but varied according to the type of CTCL (73–75% in mycosis fungoides, 90–100% in Sézary syndrome, 40–60% in lymphomatoid papulosis and 100% in other types). The positivity rate in BID was 14% with both methods. The most frequent BID with a monoclonal pattern were drug‐induced cutaneous lymphoid hyperplasia, erythrodermic psoriasis and pityriasis lichenoides chronica. Conclusions BIOMED‐2‐GS analysis of the TCRγ gene is as sensitive and specific as DGGE for CTCL diagnosis. In addition, BIOMED‐2‐GS is less time‐consuming and gives more information concerning the size and nature of TCR‐GR.     

T‐cell receptor gene rearrangement analysis of sequential biopsies in cutaneous T‐cell lymphomas with the Biomed‐2 PCR reveals transient T‐cell clones in addition to the tumor clone

Detection of a dominant T‐cell clone by T‐cell receptor (TCR) gene rearrangement analysis is often essential for the diagnosis of cutaneous T‐cell lymphomas (CTCL). The occurrence of T‐cell clones in addition to the diagnostic T‐cell clone during the course of CTCL has been reported, but the data of these studies have been contradictory. We retrospectively evaluated the data of 114 lesional skin biopsies from 26 patients with Mycosis fungoides and two patients with primary cutaneous anaplastic large cell lymphoma, which were analysed with the standardized Biomed‐2 PCR for the TCRγ and TCRβ locus. A dominant T‐cell clone was repetitively detected in 93% (26/28) of patients. Additional T‐cell clones appeared temporarily in 39% (11/28) of patients. Correlation with the clinical data did not show an association of the presence of additional T‐cell clones with age, number of treatments, progression of disease or survival. Our findings demonstrate that a persistent T‐cell clone, most likely the disease causing tumor clone, is detectable in almost all CTCL patients. In addition, transiently appearing T‐cell clones frequently occur during the course of disease. The biological relevance of these additional clones has still to be determined. However, it is important to take the possibility of additional T‐cell clones into account for diagnostic analyses.     

Genotypic analysis of cutaneous T-cell lymphomas

The gene encoding the beta-chain of the T-cell antigen receptor (TCR) has been analyzed for evidence of rearrangement in skin, blood, and lymph node specimens from 23 cases of known or suspected cutaneous T-cell lymphoma (CTCL). Two cutaneous large cell lymphomas, 4 cases of Sézary syndrome, and 5 cases of advanced (tumor) stages of mycosis fungoides showed clonal rearrangement of the TCR beta-chain gene in all samples, including lymph nodes in which histologic examination revealed only dermatopathic lymphadenitis. These results indicate that DNA analysis provides a valuable means for improving the diagnosis of extracutaneous disease in advanced stages of CTCL. In contrast, the gene was in a germline configuration in all samples from 12 patients with plaque stages of mycosis fungoides or suspected early CTCL, suggesting that in these 2 conditions the T-cell proliferation is either polyclonal or contains very few monoclonal (i.e., neoplastic) cells.     

Analyses of T-cell receptor β–chain genes by Southern blotting in known and suspected cutaneous T–cell lymphoma. A study of 67 samples from 32 patients

In this study we have investigated the configuration of the T–cell receptor (TCR) β–chain genes in benign cutaneous conditions (n= 5) and known (n= 22) or suspected (n= 5) cutaneous T–cell lymphoma (CTCL). Sequential biopsies from skin, lymph node, blood and/or bone marrow were available in 12 cases of the 22 confirmed CTCL, and a total of 67 samples were analysed. In the benign conditions, clonal rearrangements of the TCR β–chain genes were seen in neither skin nor blood samples. In contrast, in CTCL clonal rearrangements were detected in all skin samples from plaque or tumour lesions of mycosis fungoides. Clonal TCR rearrangements were also present in skin and blood samples from two patients with Sèzary's syndrome, and in skin and blood samples from three of five patients with clinically suspected CTCL. In 10 patients with large cell lymphomas, clonal rearrangements were detected in skin samples in half of the cases. In the remaining patients, clonal TCR rearrangements could not be detected in the skin, but only in the blood and/or bone marrow specimens. Results from the analyses of sequential biopsies showed identical patterns of rearrangement in 11 patients. In the remaining patient, the pattern of rearrangement differed between skin and lymph node. These data confirm and extend previous reports and indicate that analysis of TCR β–chain genes by Southern blotting forms a useful supplement to other methods for the diagnosis of known and suspected CTCL. They also emphasize the importance of studying not only skin, but also extracutaneous sites.     

Polyclonal expansion of T cells with the TCR V beta type of the tumour cell in lesions of cutaneous T-cell lymphoma: evidence for possible superantigen involvement

The involvement of superantigens in the pathology of cutaneous T-cell lymphomas (CTCL) has been suggested before, but without unequivocal evidence for superantigen activity in the patients. Seeking evidence for superantigen activity we analysed clones and microdissected single cells isolated from the epidermis of early-stage lesions of a CTCL patient for their T-cell receptor (TCR) V beta expression and TCR V gamma gene rearrangements. The vast majority of these T cells expressed the TCR V beta family type of the tumour. From their TCR gamma gene rearrangements, however, these cells were polyclonal. The tumour cell clone accounted for about 60% of these cells, about 40% were of heterogeneous origin. This dominance of a single V beta family in the polyclonally expanded dermal T-cell populations implies superantigen activity in the CTCL lesions.     

Nasal-type T/natural killer cell angiocentric lymphoma, Epstein-Barr virus-associated, and showing clonal T-cell receptor gamma gene rearrangement

Nasal-type T/natural killer (NK) cell lymphoma, which often shows an angiocentric growth pattern, is a distinct clinicopathological entity highly associated with the Epstein-Barr virus (EBV). This tumour has a characteristic immunophenotype, whereas the cytological spectrum is broad. It is known that a clonal T-cell receptor (TCR) gene rearrangement is not found in this tumour. However, it is still unresolved as to whether the finding of a clonal TCR gene rearrangement excludes the diagnosis of nasal-type T/NK cell lymphoma. We describe a case of nasal-type T/NK cell angiocentric lymphoma, EBV-associated, and showing clonal TCR gamma gene rearrangement. The patient died of sepsis 5 months after diagnosis in spite of aggressive chemotherapy.     

Immunophenotypic shift of CD4 and CD8 antigen expression in primary cutaneous T‐cell lymphomas: a clinicopathologic study of three cases

Primary cutaneous T‐cell lymphomas (CTCL) comprise a heterogeneous group of neoplasms with diverse clinical behavior. Mycosis fungoides (MF) is the most common type of CTCL. Immunophenotypical shift during progression of the disease is a rare event and its significance is unknown. We present three primary CTCL cases that showed an immunophenotypical shift and poor prognosis. Conventional hematoxylin/eosin and immunohistochemical‐stained sections were examined in all the cases. Molecular analysis for rearrangement of the T‐cell receptor (TCR) gene was performed in two cases. One case was classified as MF, while the other two lacked epidermotropism, and were considered primary cutaneous peripheral T‐cell lymphoma (PTCL), NOS. Two cases were CD3+/CD4+ and one case was CD3+/CD8+ at diagnosis. The first two patients suffered many relapses and eventually, new CTCL lesions with a CD3+/CD8+ phenotype were observed. Both cases revealed identical clonal TCR rearrangements on the initial and late lesions, supporting the interpretation of a single clonal proliferation with different phenotypes. The third case progressed with skin recurrences and pulmonary lesions with a predominant CD3+/CD4+/CD8? phenotype. All cases manifested poor prognosis and two patients died of lymphoma. Immunophenotypical shift between CD4 and CD8 in CTCL seems to be a rare phenomenon that may be associated with disease progression.     

T-cell receptor gamma gene rearrangement by multiplex polymerase chain reaction/heteroduplex analysis in patients with cutaneous T-cell lymphoma (mycosis fungoides/Sézary syndrome) and benign inflammatory disease: correlation with clinical, histological and immunophenotypical findings

BACKGROUND: A dominant T-cell clone can be detected by polymerase chain reaction (PCR) in 40-90% of cutaneous samples from patients with cutaneous T-cell lymphoma (CTCL). MATERIALS AND METHODS: From 1996 to 2003 we analysed 547 cutaneous biopsies performed to exclude CTCL (mycosis fungoides, MF/Sézary syndrome, SS). The final diagnosis was benign inflammatory disease (BID) in 353 samples (64.5%) and CTCL in 194 (35.5%). T-cell receptor (TCR)-gamma gene rearrangement was studied by using a multiplex PCR/heteroduplex (HD) analysis. The PCR results were correlated with the clinical picture, the histological pattern and the presence of T-cell lineage antigen loss, using univariate and multivariate logistic regression analyses. OBJECTIVE: To determine the sensitivity and specificity of the multiplex PCR/HD analysis and to identify which are the clinical, histopathological or immunophenotypical features significantly associated with a positive T-cell clonality. RESULTS: A clonality was demonstrated in 83.5% of CTCL and in 2.3% of BID (P < 0.001). A significantly higher percentage of clonal cases was associated with the cutaneous T-score (71.4% in T1, 76.1% in T2 and 100% in nodular and erythrodermic MF samples) and with the presence of a T-cell lineage antigen loss (93.9% vs. 77.4%). Moreover, clonality was closely related to an increase in the histopathological score (51.3% in the samples with a score < 5, compared with 92% in the lesions with > or = 5). No significant difference in the percentage of clonal cases was found between T1/T2 and T3/T4 lesions with a histopathological score > or = 5. The multivariate logistic regression showed that the density and extent of the cell infiltrate, the degree of epidermotropism and the presence of cytological atypia share an independent predictive value for clonality in T1/T2 samples, even if the highest odds ratios (3.6) were associated with the density of the cell infiltrate. The disease course of T1/T2 patients was analysed according to the PCR findings. All the PCR-negative patients showed a long-standing stable disease course; on the other hand, a disease progression occurred in 12/87 (13.8%) positive patients. CONCLUSIONS: The multiplex PCR/HD analysis is associated with a high diagnostic accuracy (92.7%) in CTCL patients. The finding of a clonal T-cell rearrangement is more closely associated with the histological pattern (in particular with the density and extent of the cell infiltrate) rather than with the MF cutaneous T-score or immunophenotype.