Methods in cell biology最新文献_第2页

In vitro culture of intestinal organoids for cell death assessments. 肠道类器官体外培养用于细胞死亡评估。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-06-02 DOI: 10.1016/bs.mcb.2026.05.001

Luying Zheng, Daichao Xu

引用次数: 0

High-throughput time-lapse imaging of B16 target cell killing by ovalbumin-specific CD8⁺ T cells. 卵清蛋白特异性CD8+ T细胞杀伤B16靶细胞的高通量延时成像。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-04-01 DOI: 10.1016/bs.mcb.2026.03.001

Beatriz Chaves, Nicolas Socquet-Juglard, Loïc Dupré, Hélène Daniels-Treffandier

{"title":"High-throughput time-lapse imaging of B16 target cell killing by ovalbumin-specific CD8+ T cells.","authors":"Beatriz Chaves, Nicolas Socquet-Juglard, Loïc Dupré, Hélène Daniels-Treffandier","doi":"10.1016/bs.mcb.2026.03.001","DOIUrl":"10.1016/bs.mcb.2026.03.001","url":null,"abstract":"Cytotoxic T Lymphocytes (CTLs) mediate tumor clearance and anti-tumoral responses through the elimination of target cancerous cells upon antigen recognition and TCR engagement. The killing of cancerous cells mediated by CTLs can be modulated by the strength of the TCR engagement, notably through the antigen affinity to the TCR, the quality of the immune synapse, the time of contact between CTLs and targets, among other parameters. Unveiling the mechanisms, kinetics and factors influencing cancer cell elimination by CTLs is essential for the amelioration of lymphocyte-based immunotherapies, such as CAR-T cells, and the understanding of tumoral clearance in vivo. Here, we provide a method to precisely monitor the cytotoxic capacity of murine CTLs in an antigen-specific manner, using ovalbumin (OVA) specific CTLs expressing the modified OT1 TCR and four different tumoral B16-F10 cell lines expressing OVA peptides with variable affinity for the OT1 TCR. We present step-by-step the implementation of a high throughput image-based pipeline allowing, on one hand, the time-lapse monitoring of CTL-mediated cytotoxicity and, on the other hand, the measurement of morphological parameters of CTLs and target cells based on fluorescent labeling. This method is suitable for ex-vivo exploration of lymphocyte cytotoxicity and related parameters, and can be adapted for mechanistic studies or screening approaches.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"207 ","pages":"119-138"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148005661","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

DNA origami-bilayer systems for dissecting the nanoscale antigen landscape in B cell immune synapse formation. DNA折纸-双层系统解剖纳米级抗原景观在B细胞免疫突触形成。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-02-04 DOI: 10.1016/bs.mcb.2026.01.020

Yuchen Hou, Chung-Yi Tseng, Raysa Orin, Emily Cohen, Leo Chou, Bebhinn Treanor

{"title":"DNA origami-bilayer systems for dissecting the nanoscale antigen landscape in B cell immune synapse formation.","authors":"Yuchen Hou, Chung-Yi Tseng, Raysa Orin, Emily Cohen, Leo Chou, Bebhinn Treanor","doi":"10.1016/bs.mcb.2026.01.020","DOIUrl":"10.1016/bs.mcb.2026.01.020","url":null,"abstract":"The recognition of antigens by B cells is the initial step in the humoral response leading to antibody production and antibody-mediated immunological memory. The majority of antigens encountered by B cells are presented in a membrane-tethered format on antigen presenting cells (APCs). Antigen-specific recognition by B cell receptors triggers the formation of an immunological synapse (IS) via a spreading and contraction response. This is critical in driving affinity discrimination and maturation by modulating the amount of antigen internalized and presented to acquire T cell help. The interaction between B cells and APCs is mimicked in vitro by planar lipid bilayers (PLBs), which have been used to reveal the importance of biophysical properties such as antigen mobility and density in triggering B cell activation. With the advent of DNA origami enabling precise control over antigen display with nanometer precision, further dissection of spatial requirements underlying IS formation is now possible. To this end, here we describe how to generate DNA origami nanogrids, attach them to PLBs in two different housing systems, and use them to stimulate B cell IS formation.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"207 ","pages":"255-281"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148005635","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

Measuring in vitro induction of antigen-specific CD8⁺ T-cell responses by murine conventional dendritic cells. 小鼠常规树突状细胞体外诱导抗原特异性CD8+ t细胞反应的测定。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-04-03 DOI: 10.1016/bs.mcb.2026.02.024

Mo D Staal, Douwe M T Bosma, Jannie Borst

引用次数: 0

Measurement and quantification of macrophage lytic cell death by kinetic microscopy. 巨噬细胞溶解细胞死亡的动力学显微镜测定和定量。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-03-25 DOI: 10.1016/bs.mcb.2026.02.019

Sara Cahill, Fiachra Humphries

{"title":"Measurement and quantification of macrophage lytic cell death by kinetic microscopy.","authors":"Sara Cahill, Fiachra Humphries","doi":"10.1016/bs.mcb.2026.02.019","DOIUrl":"10.1016/bs.mcb.2026.02.019","url":null,"abstract":"Macrophages are innate immune cells that are critical in the maintenance of tissue homeostasis and defense against pathogens. Programmed cell death is a critical tool macrophages use to clear pathogens and to alert surrounding cells to damage following induction of cell death. Diverse forms of cell death, including pyroptosis, necroptosis, ferroptosis, and secondary apoptosis following apoptosis, result in loss of plasma membrane integrity. Membrane disruption occurs following the oligomerization of pore-forming proteins into the cell membrane, releasing cytokines and damage associated molecular patterns that trigger the immune response. Thus, quantification of cell membrane permeability is an effective method for assessing cell death in macrophages. Many different factors, including macrophage polarization, stimulation, and inflammatory state, can impact macrophage predisposition to, and rate of, programmed cell death. Thus, it is important to have a method of accurately assessing macrophage cell death kinetically, rather than at a single endpoint. In this protocol, we describe a protocol for assessing cell death in macrophages by quantifying cell membrane permeability using kinetic microscopy. This method overcomes limitations of common single time point metrics for assessing cell death and is adaptable and scalable for use in assessing cell death across different cell types and treatment conditions.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"206 ","pages":"109-122"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"147817111","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

Large-scale CRISPR-Cas9 screens to define regulators of immune checkpoints. 大规模CRISPR-Cas9筛选确定免疫检查点的调节因子。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2025-11-20 DOI: 10.1016/bs.mcb.2025.10.011

Michael Mu, Johannes C Melms, Patricia Ho, Benjamin Izar

引用次数: 0

CRISPR-Cas9 screen to identify genes regulating cell death. CRISPR-Cas9筛选细胞死亡调控基因

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-06-02 DOI: 10.1016/bs.mcb.2026.05.010

Zilong Zheng, Daichao Xu

引用次数: 0

Short-term culture of human endoscopic specimens for the study of chronic inflammation. 用于慢性炎症研究的人类内镜标本的短期培养。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-06-15 DOI: 10.1016/bs.mcb.2026.05.014

Julia Werner, Laura Hennig, Laura Schiller, Albrecht Hoffmeister, Sebastian Prill, Armin Frille, Hubert Wirtz, Ngoc Anh Hoang, Florian Lordick, Cica Vissiennon, Sonja Kallendrusch

{"title":"Short-term culture of human endoscopic specimens for the study of chronic inflammation.","authors":"Julia Werner, Laura Hennig, Laura Schiller, Albrecht Hoffmeister, Sebastian Prill, Armin Frille, Hubert Wirtz, Ngoc Anh Hoang, Florian Lordick, Cica Vissiennon, Sonja Kallendrusch","doi":"10.1016/bs.mcb.2026.05.014","DOIUrl":"https://doi.org/10.1016/bs.mcb.2026.05.014","url":null,"abstract":"Inflammation and cell death are central to both chronic inflammatory disorders and cancer progression, influencing tissue homeostasis, therapeutic response, and resistance. Effective preclinical models that preserve the human microenvironment are required to study these processes, guide personalized therapy or to test pharmacologically active substances. We developed patient-derived tissue slice cultures from endoscopic specimens (ePDTC) such as gastrointestinal and pulmonary cancers, as well as inflamed mucosa from patients with Inflammatory bowel disease (IBD). These tissue cultures preserve epithelial, stromal, and immune compartments, enabling analysis of inflammatory signalling and cell death pathways while allowing parallel pharmacological testing. PDTCs maintained tissue cytoarchitecture and cell-cell interactions, permitting within-patient drug response profiling and capturing inter- and intra-patient heterogeneity. This enabled mechanistic investigation of inflammation-driven resistance and individualized assessment of chemotherapies, immunotherapies, and anti-inflammatory agents. Here, we describe a protocol for establishing ePDTC from diverse anatomical sites to provide a translational platform to study immune regulation and therapy response in human cancers and inflammatory diseases, supporting development of personalized treatment strategies.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"210 ","pages":"157-170"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"148887890","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

Flow-cytometry-based assay to study CD8⁺ T cell cytotoxicity against tumor cells. 基于流式细胞术的CD8+ T细胞对肿瘤细胞的毒性研究。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2025-11-25 DOI: 10.1016/bs.mcb.2025.10.016

Alexey V Sarapulov, Marc-Anthony Rodriguez, Roberta Zappasodi

{"title":"Flow-cytometry-based assay to study CD8+ T cell cytotoxicity against tumor cells.","authors":"Alexey V Sarapulov, Marc-Anthony Rodriguez, Roberta Zappasodi","doi":"10.1016/bs.mcb.2025.10.016","DOIUrl":"10.1016/bs.mcb.2025.10.016","url":null,"abstract":"Cancer immunotherapy approaches aim to instruct the immune system to eliminate tumor cells. Immune checkpoint blockade (ICB) therapies, such as anti-CTLA-4 and anti-PD-1, have significantly advanced the field, and other immunotherapeutic modalities are constantly being developed and tested in pre-clinical and clinical studies. One of the most important outcomes of cancer immunotherapy is efficient elimination of tumor cells by cytotoxic CD8+ T cells. Thus, accurate measurement of tumor cell sensitivity to T-cell-mediated killing is crucial for improving treatment development. This paper presents a flow cytometry-based protocol to assess antigen-specific CD8+ T-cell cytotoxicity using ovalbumin (OVA)-specific CD8+ T cells from OT-1 TCR transgenic mice and OVA-presenting tumor cell lines (either transduced with OVA or pulsed with a class-I-MHC-restricted OVA-derived peptide). This method minimizes contribution from non-antigen-mediated killing into readouts by incorporating non-target cells (not expressing/presenting-OVA cells) into co-cultures. The procedure also allows for the evaluation of T-cell-mediated cytotoxicity under various conditions and enables concurrent immunophenotyping of effector T cells and target tumor cells if desired. The protocol can be easily customized, offers advantages over methods that rely on luciferase-expressing tumor cell lines and provides versatility in detecting multiple parameters, making it a useful and valuable tool for advancing cancer immunotherapy research.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"202 ","pages":"253-276"},"PeriodicalIF":0.0,"publicationDate":"2026-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC13390104/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"146194852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

引用次数: 0

Evaluation of lipid peroxidation as a marker of ferroptosis in colon cancer cells. 评价脂质过氧化作为结肠癌细胞铁下垂的标志。

4区生物学

Methods in cell biology Pub Date : 2026-01-01 Epub Date: 2026-03-25 DOI: 10.1016/bs.mcb.2026.02.013

María Pilar Corral-Sancho, Berta Buey, Elena Layunta, Eva Latorre

引用次数: 0