RESUMO
Tumor heterogeneity complicates biomarker development and fosters drug resistance in solid malignancies. In lymphoma, our knowledge of site-to-site heterogeneity and its clinical implications is still limited. Here, we profiled 2 nodal, synchronously acquired tumor samples from 10 patients with follicular lymphoma (FL) using single-cell RNA, B-cell receptor (BCR) and T-cell receptor sequencing, and flow cytometry. By following the rapidly mutating tumor immunoglobulin genes, we discovered that BCR subclones were shared between the 2 tumor sites in some patients, but in many patients, the disease had evolved separately with limited tumor cell migration between the sites. Patients exhibiting divergent BCR evolution also exhibited divergent tumor gene-expression and cell-surface protein profiles. While the overall composition of the tumor microenvironment did not differ significantly between sites, we did detect a specific correlation between site-to-site tumor heterogeneity and T follicular helper (Tfh) cell abundance. We further observed enrichment of particular ligand-receptor pairs between tumor and Tfh cells, including CD40 and CD40LG, and a significant correlation between tumor CD40 expression and Tfh proliferation. Our study may explain discordant responses to systemic therapies, underscores the difficulty of capturing a patient's disease with a single biopsy, and furthers our understanding of tumor-immune networks in FL.
Assuntos
Evolução Clonal/genética , Linfoma Folicular/patologia , Análise de Célula Única , Adulto , Idoso , Antígenos de Neoplasias/biossíntese , Antígenos de Neoplasias/genética , Biópsia por Agulha Fina , Antígenos CD40/biossíntese , Antígenos CD40/genética , Ligante de CD40/biossíntese , Ligante de CD40/genética , DNA de Neoplasias/genética , Progressão da Doença , Feminino , Citometria de Fluxo , Rearranjo Gênico de Cadeia Leve de Linfócito B , Rearranjo Gênico do Linfócito T , Humanos , Linfonodos/química , Linfonodos/ultraestrutura , Linfócitos do Interstício Tumoral/imunologia , Linfoma Folicular/química , Linfoma Folicular/genética , Masculino , Pessoa de Meia-Idade , Proteínas de Neoplasias/biossíntese , Proteínas de Neoplasias/genética , Filogenia , RNA Neoplásico/genética , Alinhamento de Sequência , Homologia de Sequência do Ácido Nucleico , Células T Auxiliares Foliculares/imunologia , Células T Auxiliares Foliculares/metabolismo , Transcriptoma , Microambiente TumoralAssuntos
Medula Óssea/ultraestrutura , Histiocitose/etiologia , Imunoglobulinas/análise , Corpos de Inclusão/ultraestrutura , Leucoencefalopatias/etiologia , Linfonodos/ultraestrutura , Macroglobulinemia de Waldenstrom/complicações , Idoso , Cloridrato de Bendamustina/uso terapêutico , Histiócitos/ultraestrutura , Histiocitose/metabolismo , Histiocitose/patologia , Humanos , Corpos de Inclusão/química , Leucoencefalopatias/metabolismo , Leucoencefalopatias/patologia , Masculino , Plasmócitos/ultraestrutura , Síndrome , Macroglobulinemia de Waldenstrom/diagnóstico , Macroglobulinemia de Waldenstrom/tratamento farmacológico , Macroglobulinemia de Waldenstrom/patologiaRESUMO
BACKGROUND: To reveal detailed histopathological changes, virus distributions, immunologic properties and multi-omic features caused by SARS-CoV-2 in the explanted lungs from the world's first successful lung transplantation of a COVID-19 patient. MATERIALS AND METHODS: A total of 36 samples were collected from the lungs. Histopathological features and virus distribution were observed by optical microscope and transmission electron microscope (TEM). Immune cells were detected by flow cytometry and immunohistochemistry. Transcriptome and proteome approaches were used to investigate main biological processes involved in COVID-19-associated pulmonary fibrosis. RESULTS: The histopathological changes of the lung tissues were characterized by extensive pulmonary interstitial fibrosis and haemorrhage. Viral particles were observed in the cytoplasm of macrophages. CD3+ CD4- T cells, neutrophils, NK cells, γ/δ T cells and monocytes, but not B cells, were abundant in the lungs. Higher levels of proinflammatory cytokines iNOS, IL-1ß and IL-6 were in the area of mild fibrosis. Multi-omics analyses revealed a total of 126 out of 20,356 significant different transcription and 114 out of 8,493 protein expression in lung samples with mild and severe fibrosis, most of which were related to fibrosis and inflammation. CONCLUSIONS: Our results provide novel insight that the significant neutrophil/ CD3+ CD4- T cell/ macrophage activation leads to cytokine storm and severe fibrosis in the lungs of COVID-19 patient and may contribute to a better understanding of COVID-19 pathogenesis.
Assuntos
COVID-19/patologia , Hemorragia/patologia , Transplante de Pulmão , Pulmão/patologia , Linfonodos/patologia , Fibrose Pulmonar/patologia , Linfócitos B/patologia , Linfócitos B/ultraestrutura , Linfócitos B/virologia , COVID-19/genética , COVID-19/metabolismo , COVID-19/cirurgia , Cromatografia Líquida , Citometria de Fluxo , Perfilação da Expressão Gênica , Humanos , Interleucina-1beta/metabolismo , Interleucina-6/metabolismo , Células Matadoras Naturais/patologia , Células Matadoras Naturais/ultraestrutura , Células Matadoras Naturais/virologia , Pulmão/metabolismo , Pulmão/ultraestrutura , Pulmão/virologia , Linfonodos/metabolismo , Linfonodos/ultraestrutura , Linfonodos/virologia , Macrófagos Alveolares/patologia , Macrófagos Alveolares/ultraestrutura , Macrófagos Alveolares/virologia , Masculino , Pessoa de Meia-Idade , Monócitos/patologia , Monócitos/ultraestrutura , Monócitos/virologia , Neutrófilos/patologia , Neutrófilos/ultraestrutura , Neutrófilos/virologia , Óxido Nítrico Sintase Tipo II/metabolismo , Proteômica , Fibrose Pulmonar/genética , Fibrose Pulmonar/metabolismo , Fibrose Pulmonar/cirurgia , RNA-Seq , SARS-CoV-2 , Índice de Gravidade de Doença , Linfócitos T/patologia , Linfócitos T/ultraestrutura , Linfócitos T/virologia , Espectrometria de Massas em TandemRESUMO
OBJECTIVE: Measure the size and shape of talc particles in talcum powder and compare this data to the size and shape of talc particles found in surgically resected tissues from patients with ovarian carcinoma. METHODS: Using polarized light microscopy (PLM) and scanning electron microscopy (SEM), we measured the size and shape of talc particles in samples of talc-containing baby powder (TCBP) and surgically resected pelvic tissues (hysterectomies) from talc-exposed patients with ovarian carcinoma. RESULTS: The most frequent class of particles in TCBP can be unequivocally identified as talc, using both polarized light microscopy and scanning electron microscopy with energy dispersive X-ray analysis (SEM/EDX). The talc particles found in resected tissues from ovarian carcinoma patients are similar in size and shape to the most abundant morphological class of particles in TCBP. CONCLUSIONS: This finding, combined with previous epidemiological literature and tissue-based analytical studies, provides further evidence that the small, isodiametric particles that dominate TCBP can migrate from the perineum and become lodged in distal structures in the female reproductive tract, where they may lead to an increased risk of developing ovarian carcinoma.
Assuntos
Linfonodos/química , Omento/química , Ovário/química , Talco/análise , Adulto , Idoso , Carcinoma Epitelial do Ovário/patologia , Feminino , Humanos , Linfonodos/ultraestrutura , Microscopia Eletrônica de Varredura , Pessoa de Meia-Idade , Omento/ultraestrutura , Neoplasias Ovarianas/patologia , Ovário/ultraestrutura , Talco/efeitos adversos , Talco/farmacocinéticaRESUMO
Ultra-small superparamagnetic iron oxide (USPIO) nanoparticles appear to be promising tools for MR lymphography due to their unique magnetic properties. In clinical diagnosis, the effectiveness of USPIO will greatly affect the clinician's judgment to the enhanced MR images. In this study, we evaluated the effectiveness of CS015, a PAA-coated USPIO, with subcutaneous and intravenous administration. It appeared that subcutaneously injected particles had much higher efficiency to reach lymph nodes, and even worked at a very small dose 0.075 µmol/kg. Further, we compared CS015 with ferumoxytol and ferumoxtran-10 in MR lymphography and found that CS015 had the best performance. And the lymph node metastases in New Zealand rabbits were successfully detected using CS015 with one single dose. These merits of CS015 make it a promising MR lymphography contrast agent with potential applications in cancer therapy.
Assuntos
Meios de Contraste/farmacologia , Linfonodos/ultraestrutura , Linfografia/métodos , Imageamento por Ressonância Magnética/métodos , Animais , Meios de Contraste/química , Dextranos/química , Dextranos/farmacologia , Humanos , Aumento da Imagem/métodos , Linfonodos/efeitos dos fármacos , Metástase Linfática/diagnóstico por imagem , Nanopartículas de Magnetita/química , CoelhosRESUMO
INTRODUCTION: The purpose of this study was to clarify the clinicopathological features of patients with false-negative fine needle aspiration cytology (FNAC) and to determine the factors associated with negative FNAC. METHODS: Patients with negative FNAC from January 2010 to December 2019 were included. The patients with positive sentinel nodes (SN) were divided into two groups: micrometastasis (≤2 mm) group and macrometastasis (>2 mm) group. The clinicopathological characteristics were compared between the two groups using the χ2 test. RESULTS: A total of 165 patients with negative FNAC were included; 52 (31.5%) had positive SNs. Of the 52 patients, 13 (25%) had micrometastasis and the remaining 39 (75%) had macrometastasis. Of the 113 patients with negative SNs, none had metastases found in non-SNs. No significant differences were observed in age, cT stage or subtype, and preoperative ultrasound findings between the two groups. CONCLUSIONS: The false-negative rate of FNAC was high (31.5%). Micrometastatic disease was seen in patients with negative FNAC, and this might be the cause of false-negative FNAC results.
Assuntos
Biópsia por Agulha Fina , Neoplasias da Mama/diagnóstico , Linfonodos/patologia , Metástase Linfática/diagnóstico , Adulto , Idoso , Idoso de 80 Anos ou mais , Axila/diagnóstico por imagem , Axila/patologia , Neoplasias da Mama/patologia , Feminino , Humanos , Biópsia Guiada por Imagem , Linfonodos/diagnóstico por imagem , Linfonodos/ultraestrutura , Metástase Linfática/patologia , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Biópsia de Linfonodo Sentinela/métodos , UltrassonografiaRESUMO
The most common malignancy of the esophagus is squamous cell carcinoma (SCC) and regional lymph node metastases are an important prognostic factor. Isolated tumor cells (ITCs) are defined as single tumor cells or small clusters of tumor cells not exceeding 0.2 mm. The prognostic role of ITCs is not clear. This study aimed to determine the prevalence of ITCs in regional lymph nodes in patients with esophageal SCC and to investigate how frequently ITCs represent part of a true metastasis. Surgical specimens from 100 patients with SCC of the esophagus were included. All original H&E stained slides containing lymph nodes were reviewed by two gastrointestinal pathologists. In lymph nodes containing ITCs, additional levels were cut and stained with a H&E- and a cytokeratin stain. Areas of tumor cells that measured >0.2 mm on the deeper sections were classified as metastases. A total of 2460 lymph nodes were examined. ITCs were detected in 10 lymph nodes (0.4%) from nine patients (9%). Deeper sections revealed metastases in five out of the 10 lymph nodes (50%). ITCs in regional lymph nodes of patients with SCC of the esophagus is a rare finding compared with patients with adenocarcinoma of the esophagogastric junction. However, deeper sections often revealed metastases. Therefore, in patients with SCC of the esophagus, we recommend additional sectioning and immunohistochemical examination of lymph nodes when ITCs are detected on the first slide.
Assuntos
Carcinoma de Células Escamosas/patologia , Neoplasias Esofágicas/patologia , Linfonodos/patologia , Adenocarcinoma/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Escamosas/cirurgia , Estudos de Casos e Controles , Dinamarca/epidemiologia , Neoplasias Esofágicas/secundário , Junção Esofagogástrica/patologia , Feminino , Humanos , Imuno-Histoquímica/métodos , Linfonodos/metabolismo , Linfonodos/ultraestrutura , Metástase Linfática/patologia , Metástase Linfática/ultraestrutura , Masculino , Pessoa de Meia-Idade , PrognósticoRESUMO
Fibroblastic reticular cells (FRCs) are immunologically specialized myofibroblasts of lymphoid organ, and FRC maturation is essential for structural and functional properties of lymph nodes (LNs). Here we show that YAP and TAZ (YAP/TAZ), the final effectors of Hippo signaling, regulate FRC commitment and maturation. Selective depletion of YAP/TAZ in FRCs impairs FRC growth and differentiation and compromises the structural organization of LNs, whereas hyperactivation of YAP/TAZ enhances myofibroblastic characteristics of FRCs and aggravates LN fibrosis. Mechanistically, the interaction between YAP/TAZ and p52 promotes chemokine expression that is required for commitment of FRC lineage prior to lymphotoxin-ß receptor (LTßR) engagement, whereas LTßR activation suppresses YAP/TAZ activity for FRC maturation. Our findings thus present YAP/TAZ as critical regulators of commitment and maturation of FRCs, and hold promise for better understanding of FRC-mediated pathophysiologic processes.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/metabolismo , Proteínas de Ciclo Celular/metabolismo , Diferenciação Celular , Fibroblastos/metabolismo , Linfonodos/citologia , Transativadores/metabolismo , Adipócitos/metabolismo , Animais , Quimiocinas/metabolismo , Fibroblastos/ultraestrutura , Linfonodos/ultraestrutura , Receptor beta de Linfotoxina/metabolismo , Mesoderma/metabolismo , Camundongos Endogâmicos C57BL , Miofibroblastos/metabolismo , Proteínas de Sinalização YAPRESUMO
The rising incidence of cancer worldwide is causing an increase in the workload in pathology departments. This, coupled with advanced analysis methodologies, supports a developing need for techniques that could identify the presence of cancer cells in cytology and tissue samples in an objective, fast, and automated way. Fourier transform infrared (FT-IR) microspectroscopy can identify cancer cells in such samples objectively. Thus, it has the potential to become another tool to help pathologists in their daily work. However, one of the main drawbacks is the use of glass substrates by pathologists. Glass absorbs IR radiation, removing important mid-IR spectral data in the fingerprint region (1800 cm-1 to 900 cm-1). In this work, we hypothesized that, using glass coverslips of differing compositions, some regions within the fingerprint area could still be analyzed. We studied three different types of cells (peripheral blood mononuclear cells, a leukemia cell line, and a lung cancer cell line) and lymph node tissue placed on four different types of glass coverslips. The data presented here show that depending of the type of glass substrate used, information within the fingerprint region down to 1350 cm-1 can be obtained. Furthermore, using principal component analysis, separation between the different cell lines was possible using both the lipid region and the fingerprint region between 1800 cm-1 and 1350 cm-1. This work represents a further step towards the application of FT-IR microspectroscopy in histopathology departments.
Assuntos
Leucócitos Mononucleares/ultraestrutura , Linfonodos/ultraestrutura , Neoplasias/ultraestrutura , Espectroscopia de Infravermelho com Transformada de Fourier/métodos , Linhagem Celular Tumoral , Vidro/química , HumanosRESUMO
BACKGROUND: Intrapulmonary lymph nodes (IPLNs) are often recognized as sub-centimeter solid pulmonary nodules (SCPNs). The present study investigated their clinical and pathological characteristics to allow clinicians to distinguish them from malignant nodules. METHODS: Among 194 SCPNs surgically resected between 2006 and 2016, 26 IPLNs were investigated histopathologically. In addition, 145 resected malignant SCPNs were compared radiographically with the 26 IPLNs. RESULTS: Radiographically, most IPLNs were in a middle or lower lobe, and all lesions were within 20â¯mm of the visceral pleura. Enlargement was seen in one lesion. Three lesions demonstrated linear density contiguous to pleura (LD), and 13 lesions were adjacent to the peripheral pulmonary vein (APV). Microscopically, all IPLNs showed adjacency to pulmonary veins, 23 showed interlobular septa extending from the IPLN, and 18 were surrounded by a dilatated lymphoid channel. Radiographical findings of LD and APV were also seen in malignant SCPNs (LD, 12/145; APV, 25/145). Comparative analysis revealed that enlargement and APV were significant predictors differentiating IPLNs from malignant SCPNs. The sensitivity/specificity of enlargement and APV were 92%/92% and 17%/50%, respectively. CONCLUSIONS: IPLNs show typical high-resolution computed tomography findings that reflect their histopathological characteristics. Such findings help identify IPLNs prior to surgery. Specifically, enlargement and APV may differentiate IPLNs from malignant SCPNs. However, atypical cases are also possible, and radiological findings are not specific for differentiating IPLNs from malignant lesions. Thus, clinicians should consider surgical exploration when diagnosing SCPNs.
Assuntos
Linfonodos/patologia , Nódulos Pulmonares Múltiplos/diagnóstico , Nódulos Pulmonares Múltiplos/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Diagnóstico Diferencial , Feminino , Humanos , Pulmão , Linfonodos/diagnóstico por imagem , Linfonodos/ultraestrutura , Masculino , Pessoa de Meia-Idade , Nódulos Pulmonares Múltiplos/diagnóstico por imagem , Nódulos Pulmonares Múltiplos/ultraestrutura , Veias Pulmonares , Intensificação de Imagem Radiográfica , Tomografia Computadorizada por Raios XRESUMO
Changes in the structure and cell composition of carinal lymph nodes were studied in humans during aging. Replacement of node parenchyma with fibrous connective tissue progressing with age was demonstrated. The medullary matter significantly prevailed over the cortical substance. The lymph nodes in the cortical substance were small and had no light centers; the concentration of mature CD20+ B cells was high; the paracortical area was fragmented and thinned and contained no CD4+ T helpers. Ki-67+ cells were absent in all structural components of the lymph nodes reflecting exhaustion of lymphopoietic function, which was determined by the replacement of the reticular tissue of the microenvironment with the connective tissue and by the absence of CD4+ T cells regulating cellular and humoral immunity. The disintegration of the reticular stroma in the sinus system that acts as a biological filter impairs the function of lymph purification.
Assuntos
Envelhecimento/imunologia , Linfócitos B/imunologia , Tecido Conjuntivo/imunologia , Linfonodos/imunologia , Tecido Parenquimatoso/imunologia , Idoso de 80 Anos ou mais , Envelhecimento/patologia , Antígenos CD20/genética , Antígenos CD20/imunologia , Autopsia , Linfócitos B/patologia , Biomarcadores/metabolismo , Tecido Conjuntivo/patologia , Tecido Conjuntivo/ultraestrutura , Feminino , Fibrose , Expressão Gênica , Granulócitos/imunologia , Granulócitos/patologia , Humanos , Antígeno Ki-67/genética , Antígeno Ki-67/imunologia , Linfonodos/patologia , Linfonodos/ultraestrutura , Contagem de Linfócitos , Macrófagos/imunologia , Macrófagos/patologia , Masculino , Tecido Parenquimatoso/patologia , Tecido Parenquimatoso/ultraestrutura , Células Estromais/imunologia , Células Estromais/patologia , Cavidade Torácica/imunologia , Cavidade Torácica/patologiaRESUMO
Lymph node diagnostics are essentially based on cutting thin sections of formalin fixed tissues. After hematoxylin and eosin stain, Giemsa stain and immunohistochemical staining of these tissues, the lymph node diagnosis is done using a light microscope, looking at two-dimensional pictures. Three-dimensional visualizations of lymph node tissue have not been used in lymphoma diagnostics yet. This article describes three-dimensional visualization of lymphoid tissue, using thick paraffin sections, immunostained with monoclonal antibodies, confocal laser scanning and data processing with appropriate software and the 3D printing process itself. The advantages and disadvantages of different printing techniques are discussed as well as the application of 3D models in diagnostics, teaching and research of lymph nodes.
Assuntos
Linfonodos/ultraestrutura , Modelos Biológicos , Impressão Tridimensional/instrumentação , HumanosAssuntos
Leucemia Mieloide Aguda/diagnóstico , Leucemia de Células T/diagnóstico , Linfonodos/patologia , Idoso , Antineoplásicos/administração & dosagem , Antineoplásicos/uso terapêutico , Biópsia , Células da Medula Óssea/patologia , Diagnóstico Diferencial , Erros de Diagnóstico/prevenção & controle , Evolução Fatal , Técnicas Histológicas , Humanos , Leucemia Mieloide Aguda/tratamento farmacológico , Leucemia Mieloide Aguda/patologia , Leucemia de Células T/tratamento farmacológico , Leucemia de Células T/patologia , Linfonodos/ultraestrutura , Masculino , Leucemia-Linfoma Linfoblástico de Células Precursoras/diagnóstico , Leucemia-Linfoma Linfoblástico de Células Precursoras/patologiaRESUMO
Whole slide imaging (WSI) uses robotic microscopes for computerising entire slides into digital images. The aim of this study was to assess the agreement between WSI and optical microscopy for evaluating canine lymphoma cytological samples. Forty-four slides were computerised using a WSI scanner and the digital and glass slides were examined by three observers with different levels of expertise. Morphology and grade of lymphoma were scored on the basis of the updated Kiel classification and intra-observer agreement was assessed. The accuracy of determining the grade of lymphoma with digital and glass slides based on the results of flow cytometry (FC) was established. The overall intra-observer agreement for cytomorphological features was fair to moderate (κ=0.34-0.52) for the three observers and moderate (κ=0.44-0.53) for the evaluation of grade of malignancy. The diagnostic agreement between FC and digital slides was slight (κ=0.16) for the inexperienced observer, fair (κ=0.32) for the mildly experienced observer and moderate (κ=0.50) for the very experienced observer. The diagnostic agreement between FC and glass slides was fair (κ=0.37) for the inexperienced observer, substantial (κ=0.63) for the mildly experienced observer and moderate (κ=0.50) for the very experienced observer. These findings underline the importance of observer experience in determining the grade of malignancy, especially if digital slides are used. The study also identifies some technical limitations of the WSI scanner used in this study, mainly linked to image quality, which might affect the morphological evaluation of neoplastic cells.
Assuntos
Doenças do Cão/diagnóstico por imagem , Interpretação de Imagem Assistida por Computador , Linfoma/veterinária , Microscopia/veterinária , Variações Dependentes do Observador , Animais , Cães , Linfonodos/citologia , Linfonodos/ultraestrutura , Linfoma/diagnóstico por imagem , Linfoma/ultraestrutura , Microscopia/instrumentação , Microscopia/métodos , Patologia ClínicaRESUMO
PURPOSE: The lymphatic flow along the posterior gastric artery (PGA) is considered of possible clinical importance in terms of lymphatic metastasis; however, little is known about the lymph nodes (LNs) around this artery. The purpose of this study was to establish if LNs exist around the PGA and to evaluate their clinical implications. METHODS: We examined the tissues surrounding the PGA from 21 cadavers to search for LNs. We also investigated the patterns of lymphatic metastases in patients who underwent surgery for gastric neoplasms at our institute to detect their presence along the PGA. RESULTS: The PGA was identified in 11 cadavers, and LNs around the PGA were detected microscopically in 2 of these. Lymphatic metastasis directly to the LNs at the splenic artery without any metastases was regarded as skip metastasis along the PGA. Skip metastasis was found in two of ten patients who underwent surgery for remnant gastric cancer. CONCLUSIONS: The existence of LNs around the PGA was confirmed, and based on our findings, lymphatic metastasis through the PGA is possible in patients with remnant gastric cancer.
Assuntos
Artérias Epigástricas/anatomia & histologia , Linfonodos/anatomia & histologia , Linfonodos/patologia , Neoplasias Gástricas/irrigação sanguínea , Neoplasias Gástricas/patologia , Humanos , Linfonodos/ultraestrutura , Metástase Linfática , Neoplasias Gástricas/ultraestruturaRESUMO
BACKGROUND: As only a minor portion of the information present in histological sections is accessible by eye, recognition and quantification of complex patterns and relationships among constituents relies on digital image analysis. In this study, our working hypothesis was that, with the application of digital image analysis technology, visually unquantifiable breast cancer microarchitectural features can be rigorously assessed and tested as prognostic parameters for invasive breast carcinoma of no special type. METHODS: Digital image analysis was performed using public domain software (ImageJ) on tissue microarrays from a cohort of 696 patients, and validated with a commercial platform (Visiopharm). Quantified features included elements defining tumour microarchitecture, with emphasis on the extent of tumour-stroma interface. The differential prognostic impact of tumour nest microarchitecture in the four immunohistochemical surrogates for molecular classification was analysed. Prognostic parameters included axillary lymph node status, breast cancer-specific survival, and time to distant metastasis. Associations of each feature with prognostic parameters were assessed using logistic regression and Cox proportional models adjusting for age at diagnosis, grade, and tumour size. RESULTS: An arrangement in numerous small nests was associated with axillary lymph node involvement. The association was stronger in luminal tumours (odds ratio (OR) = 1.39, p = 0.003 for a 1-SD increase in nest number, OR = 0.75, p = 0.006 for mean nest area). Nest number was also associated with survival (hazard ratio (HR) = 1.15, p = 0.027), but total nest perimeter was the parameter most significantly associated with survival in luminal tumours (HR = 1.26, p = 0.005). In the relatively small cohort of triple-negative tumours, mean circularity showed association with time to distant metastasis (HR = 1.71, p = 0.027) and survival (HR = 1.8, p = 0.02). CONCLUSIONS: We propose that tumour arrangement in few large nests indicates a decreased metastatic potential. By contrast, organisation in numerous small nests provides the tumour with increased metastatic potential to regional lymph nodes. An outstretched pattern in small nests bestows tumours with a tendency for decreased breast cancer-specific survival. Although further validation studies are required before the argument for routine quantification of microarchitectural features is established, our approach is consistent with the demand for cost-effective methods for triaging breast cancer patients that are more likely to benefit from chemotherapy.
Assuntos
Neoplasias da Mama/ultraestrutura , Mama/ultraestrutura , Linfonodos/ultraestrutura , Prognóstico , Adulto , Idoso , Mama/diagnóstico por imagem , Neoplasias da Mama/diagnóstico por imagem , Neoplasias da Mama/tratamento farmacológico , Intervalo Livre de Doença , Feminino , Humanos , Processamento de Imagem Assistida por Computador , Linfonodos/diagnóstico por imagem , Metástase Linfática , Pessoa de Meia-Idade , Modelos de Riscos ProporcionaisRESUMO
Pathologists typically encounter many disparate exogenous materials in clinical specimens during their routine histopathological examinations, especially within the skin, lymph nodes, and lungs. These foreign substances may be free extracellular deposits or induce several clinical abnormalities or histopathological patterns. However, pathologists almost never investigate or report the chemical nature of exogenous metals in clinical specimens due to a lack of convenient and available technologies. In this paper, a novel strategy based on laser-induced breakdown spectroscopy (LIBS) technology is evaluated for in situ multi-elemental tissue imaging. The improved procedures allow visualization of the presence of chemical elements contained within paraffin-embedded specimens of medical interest with elemental images that are stackable with conventional histology images. We selected relevant medical situations for which the associated pathology reports were limited to the presence of lymphohistiocytic and inflammatory cells containing granules (a granuloma and a pseudolymphoma) or to lymph nodes or skin tissues containing pigments or foreign substances. Exogenous elements such as aluminum, titanium, copper, and tungsten were identified and localized within the tissues. The all-optical LIBS elemental imaging instrument that we developed is fully compatible with conventional optical microscopy used for pathology analysis. When combined with routine histopathological analysis, LIBS is a versatile technology that might help pathologists establish or confirm diagnoses for a wide range of medical applications, particularly when the nature of external agents present in tissues needs to be investigated.
Assuntos
Reação a Corpo Estranho/patologia , Espectrofotometria Atômica/métodos , Humanos , Lasers , Linfonodos/química , Linfonodos/ultraestrutura , Metais/análise , Inclusão em Parafina , Estudos Retrospectivos , Pele/química , Pele/ultraestruturaRESUMO
The extracellular matrix (ECM) is a master regulator of cellular phenotype and behavior. It has a crucial role in both normal tissue homeostasis and disease pathology. Here we present a fast and efficient approach to enhance the study of ECM composition and structure. Termed in situ decellularization of tissues (ISDoT), it allows whole organs to be decellularized, leaving native ECM architecture intact. These three-dimensional decellularized tissues can be studied using high-resolution fluorescence and second harmonic imaging, and can be used for quantitative proteomic interrogation of the ECM. Our method is superior to other methods tested in its ability to preserve the structural integrity of the ECM, facilitate high-resolution imaging and quantitatively detect ECM proteins. In particular, we performed high-resolution sub-micron imaging of matrix topography in normal tissue and over the course of primary tumor development and progression to metastasis in mice, providing the first detailed imaging of the metastatic niche. These data show that cancer-driven ECM remodeling is organ specific, and that it is accompanied by comprehensive changes in ECM composition and topological structure. We also describe differing patterns of basement-membrane organization surrounding different types of blood vessels in healthy and diseased tissues. The ISDoT procedure allows for the study of native ECM structure under normal and pathological conditions in unprecedented detail.
Assuntos
Membrana Basal/ultraestrutura , Neoplasias da Mama/ultraestrutura , Matriz Extracelular/ultraestrutura , Neoplasias Mamárias Experimentais/ultraestrutura , Proteômica , Microambiente Tumoral , Animais , Membrana Basal/metabolismo , Neoplasias da Mama/metabolismo , Matriz Extracelular/metabolismo , Feminino , Humanos , Imageamento Tridimensional , Lactação , Neoplasias Pulmonares/metabolismo , Neoplasias Pulmonares/secundário , Neoplasias Pulmonares/ultraestrutura , Linfonodos/metabolismo , Linfonodos/ultraestrutura , Metástase Linfática , Glândulas Mamárias Humanas/metabolismo , Glândulas Mamárias Humanas/ultraestrutura , Neoplasias Mamárias Experimentais/metabolismo , Melanoma Experimental/metabolismo , Melanoma Experimental/ultraestrutura , Camundongos , Imagem Óptica , Nervos Periféricos/metabolismo , Nervos Periféricos/ultraestrutura , Neoplasias da Língua/metabolismo , Neoplasias da Língua/ultraestruturaRESUMO
Parenchymal migration of naive CD4+ T cells in lymph nodes (LNs) is mediated by the Rac activator DOCK2 and PI3Kγ and is widely assumed to facilitate efficient screening of dendritic cells (DCs) presenting peptide-MHCs (pMHCs). Yet how CD4+ T cell motility, DC density, and pMHC levels interdependently regulate such interactions has not been comprehensively examined. Using intravital imaging of reactive LNs in DC-immunized mice, we show that pMHC levels determined the occurrence and timing of stable CD4+ T cell-DC interactions. Despite the variability in interaction parameters, ensuing CD4+ T cell proliferation was comparable over a wide range of pMHC levels. Unexpectedly, decreased intrinsic motility of DOCK2-/- CD4+ T cells did not impair encounters with DCs in dense paracortical networks and, instead, increased interaction stability, whereas PI3Kγ deficiency had no effect on interaction parameters. In contrast, intravital and whole-organ imaging showed that DOCK2-driven T cell motility was required to detach from pMHClow DCs and to find rare pMHChigh DCs. In sum, our data uncover flexible signal integration by scanning CD4+ T cells, suggesting a search strategy evolved to detect low-frequency DCs presenting high cognate pMHC levels.
Assuntos
Linfócitos T CD4-Positivos/imunologia , Comunicação Celular , Movimento Celular , Células Dendríticas/imunologia , Proteínas Ativadoras de GTPase/metabolismo , Animais , Apresentação de Antígeno , Classe Ib de Fosfatidilinositol 3-Quinase/deficiência , Classe Ib de Fosfatidilinositol 3-Quinase/genética , Classe Ib de Fosfatidilinositol 3-Quinase/metabolismo , Proteínas Ativadoras de GTPase/deficiência , Proteínas Ativadoras de GTPase/genética , Fatores de Troca do Nucleotídeo Guanina , Antígenos de Histocompatibilidade/imunologia , Microscopia Intravital/métodos , Linfonodos/imunologia , Linfonodos/ultraestrutura , Ativação Linfocitária , CamundongosRESUMO
A significantly increased incidence of pleural mesothelioma in Biancavilla (Sicily, Italy) has been attributed to exposure to fluoro-edenite (FE), a fibrous amphibole extracted from a local stone quarry. The lymph-nodes draining the pulmonary lobes of sheep grazing around the town were examined, to gain insights into fibre diffusion. The pasture areas of six sheep flocks lying about 3km from Biancavilla were located using the global positioning system. The cranial tracheobronchial and one middle mediastinal lymph-node as well as four lung tissue samples were collected from 10 animals from each flock and from 10 control sheep for light and scanning electron microscopy (SEM) examination. The lymph-nodes from exposed sheep were enlarged and exhibited signs of anthracosis. Histologically, especially at the paracortical level, they showed lymph-follicle hyperplasia with large reactive cores and several macrophages (coniophages) containing grey-brownish particulate interspersed with elements with a fibril structure, forming aggregates of varying dimensions (coniophage nodules). Similar findings were detected in some peribronchiolar areas of the lung parenchyma. SEM examination showed that FE fibres measured 8-41µm in length and 0.4-1.39µm in diameter in both lymph-nodes and lung tissue. Monitoring of FE fibres in sheep lymph-nodes using appropriate techniques can help set up environmental pollution surveillance.