dual immunohistochemistry

  • 文章类型: Journal Article
    2型神经纤维瘤病(NF2)丢失发生在大约30%至50%的弥漫性胸膜间皮瘤(DPM)中,并在肿瘤核中积累了与PDZ结合基序(TAZ)相关的蛋白(YAP)1和转录共激活因子。NF2和YAP/TAZ代表潜在的治疗靶标。我们研究了NF2-YAP/TAZ双重免疫组织化学(IHC)在识别具有NF2改变的DPM以及将DPM与良性间皮增殖区分开方面的性能。随后在具有(n=10)或不具有(n=10)通过下一代测序(NGS)检测到的NF2改变和9例良性病例的DPM的发现队列中进行NF2-YAP/TAZIHC。在Discovery群组中确定NF2表达丧失和使用IHC的YAP/TAZ过表达的截断值。在验证队列(20例DPM和10例良性病例)中研究了NF2-YAP/TAZIHC的性能特征。在“发现”队列中,使用NGS进行NF2改变的所有DPM均显示NF2IHC评分<2,而所有NF2野生型DPM均显示评分≥2.NF2改变的DPM的YAP/TAZH评分(P<.001)显着高于NF2野生型DPM和良性胸膜(中位数H评分:237.5[范围,185-275],130.0[范围,40-225],和10.0[范围,0-75],分别)。NF2-YAP/TAZIHC显示95.2%的敏感性,100%特异性,100%阳性预测值,以NGS为金标准,检测DPM中NF2改变(n=40)的阴性预测值为95%,将DPM(n=40)与良性间皮增殖(n=19)区分为87.5%的敏感性和100%的特异性。NF2-YAP/TAZIHC对检测DPM中的NF2改变具有高灵敏度和特异性,对恶性肿瘤具有高特异性,强调指导NF2靶向治疗和区分DPM与良性模拟的潜在效用。
    Neurofibromatosis type 2 (NF2) loss occurs in approximately 30% to 50% of diffuse pleural mesothelioma (DPM) with accumulation of yes-associated protein (YAP) 1 and transcriptional coactivator with PDZ-binding motif (TAZ) in tumor nuclei. NF2 and YAP/TAZ represent potential therapeutic targets. We investigated the performance of NF2-YAP/TAZ dual immunohistochemistry (IHC) in identifying DPM that harbors NF2 alterations and in distinguishing DPM from benign mesothelial proliferations. NF2-YAP/TAZ IHC was subsequently performed in a Discovery cohort of DPMs with (n = 10) or without (n = 10) NF2 alterations detected by next-generation sequencing (NGS) and 9 benign cases. The cutoff values for loss of NF2 expression and YAP/TAZ overexpression using IHC were determined in the Discovery cohort. The performance characteristics of NF2-YAP/TAZ IHC were investigated in a Validation cohort (20 DPMs and 10 benign cases). In the Discovery cohort, all DPMs with NF2 alterations using NGS showed NF2 IHC scores of <2, whereas all NF2-wild-type DPMs showed scores of ≥2. NF2-altered DPMs had significantly higher YAP/TAZ H-scores (P < .001) than NF2-wild-type DPM and benign pleura (median H-scores: 237.5 [range, 185-275], 130.0 [range, 40-225], and 10.0 [range, 0-75], respectively). NF2-YAP/TAZ IHC demonstrated 95.2% sensitivity, 100% specificity, 100% positive predictive value, and 95% negative predictive value for detecting NF2 alterations in DPM (n = 40) with NGS as the gold standard and 87.5% sensitivity and 100% specificity for distinguishing DPM (n = 40) from benign mesothelial proliferations (n = 19). NF2-YAP/TAZ IHC has a high sensitivity and specificity for detecting NF2 alterations in DPM and a high specificity for malignancy, highlighting potential utility for guiding NF2-targeted therapies and distinguishing DPM from benign mimics.
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  • 文章类型: Journal Article
    口腔鳞状细胞癌(OSCC)是影响口腔颌面部区域的最常见癌症。本研究旨在探讨肿瘤干细胞(CSCs)在OSCC血管生成和缺氧反应中的作用。
    这项回顾性观察性研究使用双重免疫组织化学评估了56例OSCC。使用八聚体结合转录因子3/4(0CT3/4)标记来评估CSC活性。葡萄糖转运蛋白1(GLUT1)标记物用于评估缺氧反应和血管生成,而endoglin(CD105)用于评估血管生成和血管形成的晚期阶段。
    在12例III级OSCC患者中的11例观察到OCT3/4和GLUT1的共表达。然而,我们在22例I级OSCC患者中的13例未观察到这些标志物的共表达.尽管我们观察到GLUT1和OCT3/4的共表达与肿瘤分级之间存在显着相关性,OCT3/4和CD105的共表达与不同程度的OSCC之间没有显着相关性。
    CSCs在缺氧反应和血管生成的初始阶段可能发挥重要作用。我们的结果显示,在较高等级的OSCC中,GLUT1作为对缺氧情况的第一反应可能是CSC的结果。需要进一步的研究来发现其他生物标志物,他们的角色,以及OSCC中CSCs的相关通路。
    Oral squamous cell carcinoma (OSCC) is the most common cancer affecting the oral and maxillofacial region. This study aimed to investigate the role of cancer stem cells (CSCs) in angiogenesis and hypoxic response in OSCC.
    This retrospective observational study evaluated 56 cases of OSCC using dual immunohistochemistry. Octamer-binding transcription factor 3/4 (OCT3/4) marker was used to evaluate CSC activity. Glucose transporter 1 (GLUT1) marker was used to evaluate the hypoxic response and angiogenesis, while endoglin (CD105) was used to evaluate the late stage of angiogenesis and blood vessel formation.
    Co-expression of OCT3/4 and GLUT1 was noted in 11 of 12 patients with grade III OSCC. However, we did not observe co-expression of these markers in 13 of 22 patients with grade I OSCC. Although we observed a significant correlation between co-expression of GLUT1 and OCT3/4 and tumor grade, there was no significant correlation between co-expression of OCT3/4 and CD105 and different grades of OSCC.
    CSCs could play important roles in the initial stages of hypoxic response and angiogenesis. Our result reported that in higher grades of OSCC, GLUT1 as a first response to hypoxic situations might be a result of CSCs. Further studies are required to discover other biomarkers, their roles, and associated pathways of CSCs in OSCC.
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  • 文章类型: Journal Article
    Pilomatricoma是一种相对常见的良性皮肤附件肿瘤和公认的实体,而其色素变体则不那么常见,报道也较少。其估计频率范围从11%到24%,根据有限数量的出版案例系列。本文介绍了一名42岁男子的病例,该男子表现出乳晕周围区域的牢固皮下结节。组织病理学检查显示囊性病变由基质和基质上细胞组成,并伴有异物肉芽肿细胞反应。有趣的是,可检测到色素沉着过度的区域,有许多增生的黑素细胞和很少的有丝分裂。为了评估色素沉着区有丝分裂活性成分的细胞谱系,使用Ki67/Mart1和p63/SOX10进行双重免疫组织化学。色素性绒毛瘤是一种被低估的,漏报变体,和双重免疫组织化学染色是提供正确解释不同细胞群体中增殖活性的有效工具。
    Pilomatricoma is a relatively common benign cutaneous adnexal tumor and a well-recognized entity, while its pigmented variant is far less common and less reported. Its estimated frequency ranges from 11 to 24%, according to a limited number of published case series. This article describes the case of a 42-year-old man presenting a firm subcutaneous nodule of the periareolar region. Histopathologic examination revealed a cystic lesion composed of matrical and supramatrical cells accompanied by a foreign body granulomatous cell reaction. Interestingly, a hyperpigmented area with numerous hyperplastic melanocytes and few mitoses was detectable. In order to assess the cell lineage of the mitotically active component in the hyperpigmented area, double immunohistochemistry with Ki67/Mart1 and p63/SOX10 was performed. Pigmented pilomatricoma is an underrecognized, underreported variant, and double immunohistochemistry stain is an effective tool in providing the correct interpretation of the proliferative activity in the different cellular populations.
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  • 文章类型: Journal Article
    我们回顾了341例边缘区淋巴瘤(MZL)(47)或滤泡性淋巴瘤(FL)(294)的连续病例,其中7例由于滤泡灶中肿瘤细胞对BCL6和BCL2的共表达而难以区分。这刺激了我们开发双重BCL6/BCL2免疫组织化学,允许我们评估单个细胞之间的共表达。双重染色证实7例中有6例共表达,所有结外MZL(ENMZL)基于整体特征,占本系列MZL的13%。这些发现证实了MZL细胞在生发中心(GC)微环境中的蛋白质表达具有可塑性,一个重要的诊断缺陷。有趣的是,在所有表达BCL6的MZL中,定植卵泡内的非肿瘤性GCB细胞显示CD10表达减少或缺失,但保留了BCL6和高ki67。这一发现表明,在MZL定植的背景下,非肿瘤性GCB细胞中CD10表达的可塑性,可能与NF-kB失调有关。
    We reviewed 341 consecutive cases of marginal zone lymphoma (MZL) (47) or follicular lymphoma (FL) (294) of which 7 were difficult to distinguish due to perceived coexpression of BCL6 and BCL2 by tumor cells in follicular foci. This stimulated us to develop dual BCL6/BCL2 immunohistochemistry, allowing us to assess coexpression among individual cells. Dual staining confirmed coexpression in 6 of 7 cases, all extranodal MZL (ENMZL) based on overall features and representing 13% of MZL in this series. These findings confirm that MZL cells have plasticity regarding protein expression within the germinal center (GC) microenvironment, an important diagnostic pitfall. Intriguingly, in all MZL expressing BCL6, non-neoplastic GC B cells within colonized follicles showed diminished or absent CD10 expression but preserved BCL6 and high ki67. This finding suggests plasticity of CD10 expression in non-neoplastic GC B cells in the context of colonization by MZL, possibly related to NF-kB dysregulation.
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  • 文章类型: Journal Article
    背景:淋巴管浸润(LVI)被认为是黑素瘤细胞可以扩散到区域淋巴结和远处部位的机制,并且可能是不良结果的预测因素。在苏木精-伊红(HE)染色的切片上通常很难检测到淋巴管浸润,容易用双重免疫组织化学(IHC)鉴定黑素细胞和血管标志物。
    方法:纳入100例Breslow厚度为1-4mm且常规HE评估缺乏LVI的原发性皮肤恶性黑色素瘤病例。我们比较了CD31/S100和CD34/S100以及D2-40/S100双重染色的LVI检出率,并检查了LVI与临床结果的关联。
    结果:CD31/S100,CD34/S100和D2-40/S100的双重免疫组织化学阳性为40(40%),17(17%)和35(35%),分别。在多变量分析中,LVI是SLN状态的独立预测因子。多因素分析显示,LVI和男性是影响总生存的独立危险因素。
    结论:双重IHC改善了LVI的识别,并预测了SLN转移。使用双IHC检测LVI,特别是通过CD31/S100和D2-40/S100的组合是一个有用的步骤,建议将其纳入皮肤黑色素瘤的基本评估参数中。
    BACKGROUND: Lymphovascular invasion (LVI) is believed to be the mechanism by which melanoma cells can disseminate to regional lymph nodes and distant sites and may be predictive of adverse outcome. Lymphovascular invasion often difficult to detect on hematoxylin-eosin (HE) stained sections, are readily identified with dual immunohistochemistry (IHC) for melanocytic and vascular markers.
    METHODS: A total of 100 primary cutaneous malignant melanoma cases that had a Breslow thickness of 1-4 mm and lacked LVI by conventional HE assessment were included. We compared the LVI detection rates of double staining for CD31/S100 and CD34/S100, and D2-40/S100, and examined the association of LVI with clinical outcomes.
    RESULTS: The dual immunohistochemical positivity for CD31/S100, CD34/S100, and D2-40/S100 were 40(40%), 17(17%) and 35(35%), respectively. On multivariate analysis, LVI was an independent predictor of SLN status. Multivariate analysis revealed that LVI and male gender were independent risk factors for overall survival.
    CONCLUSIONS: The recognition of LVI is improved by dual IHC and predicts SLN metastasis. The detection of LVI using dual IHC, especially by a combination of CD31/S100 and D2-40/S100 is a useful step that inclusion should be recommended in basic evaluation parameters for cutaneous melanoma.
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  • 文章类型: Journal Article
    Immunohistochemistry is an integral technique for tissue-based diagnostics and biomarker detection with broad worldwide adoption. Advances in core chemistries, antibody design, and automation have ushered unprecedented sensitivity, specificity, and reproducibility in immunohistochemistry assays. As a result, clinical immunohistochemistry assays that utilize dual-color approaches and mutation-specific antibodies provide novel tools in clinical diagnostics that until recently were in the realm of investigational research. This review provides an overview of innovations in clinical immunohistochemistry assays with emphasis on those used for patients with hematopoietic neoplasms.
    Advances in clinical-grade immunohistochemistry techniques have allowed labs to develop and validate multiplex assays that improve diagnostic utility-such as CD5/PAX5 and TCF4/CD123 dual-color stains-and have the potential to enhance the specificity of biomarker detection. In addition, the increased availability of immunohistochemistry assays that detect mutant proteins (e.g., BRAF V600E and IDH1 R132H) provides a helpful replacement and/or adjunct for molecular testing. These techniques are highly reproducible, entail reasonable technical and interpretation complexity, and are relatively cost-effective, making them valuable novel tools in modern cancer care. Multiplex and mutation-specific immunohistochemistry assays represent important innovations that provide improved utility in the context of personalized medicine and targeted therapy.
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  • 文章类型: Journal Article
    Immunohistochemistry on mouse tissue utilizing mouse monoclonal antibodies presents a challenge. Secondary antibodies directed against the mouse monoclonal primary antibody of interest will also detect endogenous mouse immunoglobulin in the tissue. This can lead to significant spurious staining. Therefore, a \"mouse-on-mouse\" staining strategy is needed to yield credible data. This paper presents a method that is easy to use and highly flexible to accommodate both an avidin-biotin detection system as well as a biotin-free polymer detection system. The mouse primary antibody is first combined with an Fab fragment of an anti-mouse antibody in a tube and allowed sufficient time to form an antibody complex. Any non-complexed secondary antibody is bound up with mouse serum. The mixture is then applied to the tissue. The flexibility of this method is confirmed with the use of different anti-mouse antibodies followed by a variety of detection reagents. These techniques can be used for immunohistochemistry (IHC), immunofluorescence (IF), as well as staining with multiple primary antibodies. This method has also been adapted to other models, such as using human antibodies on human tissue and using multiple rabbit antibodies in dual immunofluorescence.
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