Methods in cell biology最新文献_第10页

A Hitchhiker's guide to high-dimensional tissue imaging with multiplexed ion beam imaging. 利用多路离子束成像技术进行高维组织成像的 "搭便车指南"。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-10 DOI: 10.1016/bs.mcb.2024.02.018

Yao Yu Yeo, Precious Cramer, Addison Deisher, Yunhao Bai, Bokai Zhu, Wan-Jin Yeo, Margaret A Shipp, Scott J Rodig, Sizun Jiang

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Degranulation assay to evaluate NK cell natural and antibody-dependent cell-mediated cytotoxicity against A549 tumor spheroids. 脱颗粒试验评估 NK 细胞对 A549 肿瘤球的天然细胞毒性和抗体依赖性细胞介导的细胞毒性。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-07-02 DOI: 10.1016/bs.mcb.2024.06.003

Ainara Lopez-Pardo, Ainhoa Amarilla-Irusta, Víctor Sandá, Mario Stan-Fontoba, Francisco Borrego, Laura Amo

{"title":"Degranulation assay to evaluate NK cell natural and antibody-dependent cell-mediated cytotoxicity against A549 tumor spheroids.","authors":"Ainara Lopez-Pardo, Ainhoa Amarilla-Irusta, Víctor Sandá, Mario Stan-Fontoba, Francisco Borrego, Laura Amo","doi":"10.1016/bs.mcb.2024.06.003","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.06.003","url":null,"abstract":"Adoptive natural killer (NK) cell-based immunotherapy is a promising treatment approach in cancer that is showing notable efficacy against hematological malignancies. However, the success of NK cell immunotherapy in patients with solid tumors is limited due to several barriers, which include the immunosuppressive tumor microenvironment (TME), heterogeneity of tumor cells and poor NK cell infiltration into the tumor. Advances in 3D in vitro culture technologies have opened new avenues for the development of more physiological human cancer models that mimic important tumor features which are absent in traditional 2D studies and may be essential for the improvement of immunotherapies against solid tumors. Here, we describe a comprehensive protocol to generate tumor spheroids from the A549 lung carcinoma cell line, then establish co-cultures with NK cells to, ultimately, determine NK cell functional response with a degranulation assay, a surrogate of NK cell cytotoxicity against tumor spheroids. Additionally, we studied degranulation by stimulating NK cell antibody-dependent cell-mediated cytotoxicity (ADCC) with cetuximab, an IgG1 monoclonal antibody used in cancer therapy. Likewise, other monoclonal antibodies or combination treatments could also be studied in this 3D co-culture system, providing very valuable information to define effective combinations of therapeutic agents able to generate NK cells with high cytotoxic potential that could lead to more successful adoptive NK cell-based therapies for the treatment of solid tumors.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406537","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Cytofluorometric assessment of calreticulin exposure on CD38⁺ plasma cells from the human bone marrow. 细胞荧光测定法评估人骨髓 CD38+ 浆细胞上的钙网蛋白暴露。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-06-10 DOI: 10.1016/bs.mcb.2024.05.009

Manuel Beltrán-Visiedo, Alfonso Serrano-Del Valle, Nelia Jiménez-Aldúan, Ruth Soler-Agesta, Javier Naval, Lorenzo Galluzzi, Isabel Marzo

{"title":"Cytofluorometric assessment of calreticulin exposure on CD38+ plasma cells from the human bone marrow.","authors":"Manuel Beltrán-Visiedo, Alfonso Serrano-Del Valle, Nelia Jiménez-Aldúan, Ruth Soler-Agesta, Javier Naval, Lorenzo Galluzzi, Isabel Marzo","doi":"10.1016/bs.mcb.2024.05.009","DOIUrl":"10.1016/bs.mcb.2024.05.009","url":null,"abstract":"Exposure of the endoplasmic reticulum chaperone calreticulin (CALR) on the surface of stressed and dying cells is paramount for their effective engulfment by professional antigen-presenting cells such as dendritic cells (DCs). Importantly, this is required (but not sufficient) for DCs to initiate an adaptive immune response that culminates with an effector phase as well as with the establishment of immunological memory. Conversely, the early exposure of phosphatidylserine (PS) on the outer layer of the plasma membrane is generally associated with the rapid engulfment of stressed and dying cells by tolerogenic macrophages. Supporting the clinical relevance of the CALR exposure pathway, the spontaneous or therapy-driven translocation of CALR to the surface of malignant cells, as well as intracellular biomarkers thereof, have been associated with improved disease outcome in patients affected by a variety of neoplasms, with the notable exception of multiple myeloma (MM). Here, we describe an optimized protocol for the flow cytometry-assisted quantification of surface-exposed CALR and PS on CD38+ plasma cells from the bone marrow of patients with MM. With some variations, we expect this method to be straightforwardly adaptable to the detection of CALR and PS on the surface of cancer cells isolated from patients with neoplasms other than MM.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"142406536","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Severe acute respiratory syndrome coronavirus 2 (SARS-CoV2) culture and sample preparation for correlative light electron microscopy. 严重急性呼吸系统综合征冠状病毒 2（SARS-CoV2）培养和相关光电子显微镜样本制备。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2024-03-04 DOI: 10.1016/bs.mcb.2024.02.031

Maximilian Erdmann, Lorna Hodgson, Isobel Webb, Andrew D Davidson, Paul Verkade

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Humanized mouse models for anti-cancer therapy. 用于抗癌治疗的人源化小鼠模型。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-09-09 DOI: 10.1016/bs.mcb.2023.06.002

Maria Francesca Baietti, Eleonora Leucci

引用次数: 0

Identification of TCR repertoire patterns linked with anti-cancer immunotherapy. 确定与抗癌免疫疗法相关的 TCR 重排模式。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-09-23 DOI: 10.1016/bs.mcb.2023.05.001

Romi Vandoren, Sofie Gielis, Kris Laukens, Pieter Meysman

{"title":"Identification of TCR repertoire patterns linked with anti-cancer immunotherapy.","authors":"Romi Vandoren, Sofie Gielis, Kris Laukens, Pieter Meysman","doi":"10.1016/bs.mcb.2023.05.001","DOIUrl":"10.1016/bs.mcb.2023.05.001","url":null,"abstract":"The highly diverse T cell receptor (TCR) repertoire is a crucial component of the adaptive immune system that aids in the protection against a wide variety of pathogens. This TCR repertoire, comprising the collection of all TCRs in an individual, is a valuable source of information on both recent and ongoing T cell activation. Cancer cells, like pathogens, have the ability to trigger an adaptive immune response. However, because cancer cells use a variety of strategies to escape immune responses, this is often insufficient to completely eradicate them. As a result, immunotherapy is a promising treatment option for cancer patients. This treatment is expected to increase T cell activation and subsequently alter the TCR repertoire composition in these patients. Monitoring TCR repertoires before and after immunotherapy can therefore provide additional insight into T cell responses and might identify cancer-associated TCR sequences. Here we present a computational strategy to identify those changes in the TCR repertoire that occur after treatment with immunotherapy. Since this method allows the identification of TCR patterns that might be treatment-associated, it can help future research by revealing those patterns that are related with response. This TCR analysis workflow is illustrated using public data from three different cancer patients who received anti-PD-1 treatment.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140318632","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Methods behind oncolytic virus-based DC vaccines in cancer: Toward a multiphase combined treatment strategy for Glioblastoma (GBM) patients. 基于溶瘤病毒的直流电疫苗治疗癌症的方法：针对胶质母细胞瘤（GBM）患者的多阶段综合治疗策略。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-10-19 DOI: 10.1016/bs.mcb.2023.06.001

Stefaan W Van Gool, Peter Van de Vliet, Linde F C Kampers, Jennifer Kosmal, Tobias Sprenger, Ella Reich, Volker Schirrmacher, Wilfried Stuecker

引用次数: 0

Training of epitope-TCR prediction models with healthy donor-derived cancer-specific T cells. 用健康供体来源的癌症特异性 T 细胞训练表位-TCR 预测模型。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-09-15 DOI: 10.1016/bs.mcb.2023.08.001

Donovan Flumens, Sofie Gielis, Esther Bartholomeus, Diana Campillo-Davo, Sanne van der Heijden, Maarten Versteven, Hans De Reu, Evelien Smits, Benson Ogunjimi, Kris Laukens, Pieter Meysman, Eva Lion

{"title":"Training of epitope-TCR prediction models with healthy donor-derived cancer-specific T cells.","authors":"Donovan Flumens, Sofie Gielis, Esther Bartholomeus, Diana Campillo-Davo, Sanne van der Heijden, Maarten Versteven, Hans De Reu, Evelien Smits, Benson Ogunjimi, Kris Laukens, Pieter Meysman, Eva Lion","doi":"10.1016/bs.mcb.2023.08.001","DOIUrl":"10.1016/bs.mcb.2023.08.001","url":null,"abstract":"Discovery of epitope-specific T-cell receptors (TCRs) for cancer therapies is a time consuming and expensive procedure that usually requires a large amount of patient cells. To maximize information from and minimize the need of precious samples in cancer research, prediction models have been developed to identify in silico epitope-specific TCRs. In this chapter, we provide a step-by-step protocol to train a prediction model using the user-friendly TCRex webtool for the nearly universal tumor-associated antigen Wilms' tumor 1 (WT1)-specific TCR repertoire. WT1 is a self-antigen overexpressed in numerous solid and hematological malignancies with a high clinical relevance. Training of computational models starts from a list of known epitope-specific TCRs which is often not available for new cancer epitopes. Therefore, we describe a workflow to assemble a training data set consisting of TCR sequences obtained from WT137-45-reactive CD8 T cell clones expanded and sorted from healthy donor peripheral blood mononuclear cells.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"140318640","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Assessing microbiota composition in the context of aging. 评估衰老背景下的微生物群组成。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-02-08 DOI: 10.1016/bs.mcb.2022.12.007

Diana Campos-Iglesias, José M P Freije, Carlos López-Otín

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Detection of radiation-induced senescence by the Debacq-Chainiaux protocol: Improvements and upgrade in the detection of positive events. 用德巴克-凯尼奥（Debacq-Chainiaux）方案检测辐射引起的衰老：阳性事件检测的改进和升级。

4区生物学

Methods in cell biology Pub Date : 2024-01-01 Epub Date: 2023-07-11 DOI: 10.1016/bs.mcb.2022.10.015

V Paget, O Guipaud, A François, F Milliat

{"title":"Detection of radiation-induced senescence by the Debacq-Chainiaux protocol: Improvements and upgrade in the detection of positive events.","authors":"V Paget, O Guipaud, A François, F Milliat","doi":"10.1016/bs.mcb.2022.10.015","DOIUrl":"10.1016/bs.mcb.2022.10.015","url":null,"abstract":"Senescent cells are blocked in the cell cycle but remain metabolically active. These cells, once engaged in the senescence process, fail to initiate DNA replication. Due to the shortening of telomeres, replicative senescence can be triggered by a DNA damage response. Moreover, cells can also be induced to senesce by DNA damage in response to elevated reactive oxygen species (ROS), activation of oncogenes, cell-cell fusion or after ionizing radiation. There are multiple experimental ways to detect senescent cells directly or indirectly. Senescence-associated cellular traits (SA β-Gal activity, increase in cell volume and lysosome content, appearance of γ-H2AX foci, increase of ROS and oxidative damage adducts, etc.) can be identified by numerous methods of detection (flow cytometry, confocal imaging, in situ staining, etc.). Here, we improved an existing flow cytometry protocol and further developed a new one specifically tailored to ionizing radiation-induced endothelial senescence. Thus, we have upgraded the Debacq-Chainiaux protocol and added improvements in this protocol (i) to better detect positive events (ii) to offer a compatibility to simultaneously analyze various intracellular molecules including phosphorylated signaling proteins and cytokines, whether related or not to senescence processes.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":null,"pages":null},"PeriodicalIF":0.0,"publicationDate":"2024-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139672169","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0