Methods in cell biology最新文献

Evaluating amyloid-beta aggregation and toxicity in transgenic Caenorhabditis elegans models of Alzheimer's disease. 评估转基因秀丽隐杆线虫阿尔茨海默病模型中的淀粉样蛋白聚集和毒性。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-07-09 DOI: 10.1016/bs.mcb.2024.06.001

Leticia Priscilla Arantes, Larissa Marafiga Cordeiro, Félix Alexandre Antunes Soares

引用次数: 0

The contribution of automated cytometry in immuno-oncology. 自动细胞术在免疫肿瘤学中的贡献。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2023-07-17 DOI: 10.1016/bs.mcb.2023.03.005

Andrea Sbrana, Giuliano Mazzini, Giuditta Comolli, Andrea Antonuzzo, Marco Danova

{"title":"The contribution of automated cytometry in immuno-oncology.","authors":"Andrea Sbrana, Giuliano Mazzini, Giuditta Comolli, Andrea Antonuzzo, Marco Danova","doi":"10.1016/bs.mcb.2023.03.005","DOIUrl":"10.1016/bs.mcb.2023.03.005","url":null,"abstract":"Cancer immunotherapy has been a real revolution and has given many survival chances to several patients. However, the understanding of resistance to immunotherapy is still an unmet need in clinical practice. Monitoring of immune mechanisms could be a tool to better understand this phenomenon. FCM and CyTOF could be used in this field, since they allow the simultaneous analysis of several protein expressions pattern, thus possibly understanding the functions of several immune cell populations, such as T cells, and their interactions with tumor cells and tumor microenvironment. Furthermore, automated cytometry could be used to understand the interaction of drugs with their target through the analysis of receptor occupancy. Spectral overlap, however, could be a limit for multiple simultaneous analyses. Other possible limitations of these techniques are a low number of cells in samples and the need for viable cells (with the possible interference of cell debris). The lack of standardized protocols, and thus the difficult reproducibility, have been the major limit to their application in clinical practice, so international efforts have been made to get to shared guidelines. Ongoing trials are to answer to the possibility of clinical application of these techniques.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"195 ","pages":"23-37"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143780456","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

Human and mouse iNKT cell expansion and engineering with viral vectors. 人和小鼠iNKT细胞的扩增和病毒载体工程。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-02-26 DOI: 10.1016/bs.mcb.2025.01.008

Gloria Delfanti, Paolo Dellabona, Giulia Casorati, Michela Consonni

{"title":"Human and mouse iNKT cell expansion and engineering with viral vectors.","authors":"Gloria Delfanti, Paolo Dellabona, Giulia Casorati, Michela Consonni","doi":"10.1016/bs.mcb.2025.01.008","DOIUrl":"https://doi.org/10.1016/bs.mcb.2025.01.008","url":null,"abstract":"Invariant natural killer T (iNKT) cells are a non-conventional T-cell population characterized by the expression of a conserved semi-invariant T-cell receptor (TCR) with specificity towards self or microbial lipid antigens, presented by the non-polymorphic MHC class I-related molecule CD1d. iNKT cells play a pivotal role in tumor immunosurveillance and serve as a potent tool for anti-cancer therapies. Notably, iNKT cells can be effectively redirected against both hematological and solid malignancies through genetic engineering using either Chimeric Antigen Receptors (CARs) or TCRs targeted to tumor antigens. However, due to their low frequency, iNKT cell expansion in vitro is an essential step to obtain suitable cell number for adoptive cell therapy (ACT). Here we describe two robust methods for efficiently isolating primary mouse and human iNKT cells that can be easily genetically modified and expanded. iNKT cells are isolated from the spleens of iVα14-Jα18 transgenic mice or from human buffy coats resulting in highly enriched populations. Both mouse and human iNKT cells are activated with anti-CD3/CD28 beads, IL-2 and IL-7, and subsequently transduced with tumor-specific receptors, yielding millions of ready-to-use, highly pure, and stably transduced tumor-redirected iNKT cells. The final cell product is suitable for in vitro investigation of iNKT cell activation and function mechanisms, as well as for pre-clinical ACT studies.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"196 ","pages":"67-85"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667970","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

Splenocytes antitumor cytotoxicity assessment after prophylactic vaccination or drug treatment of tumor-bearing mice. 荷瘤小鼠预防性接种或药物治疗后脾细胞抗肿瘤细胞毒性评价。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-19 DOI: 10.1016/bs.mcb.2024.10.011

Kenny Misael Calvillo-Rodriguez, Ana Luisa Rivera-Lazarin, Reyes Tamez-Guerra, Ana Carolina Martinez-Torres, Cristina Rodriguez-Padilla

引用次数: 0

The evolution of flow cytometry with respect to cancer. 流式细胞术在癌症方面的发展。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-12-12 DOI: 10.1016/bs.mcb.2024.11.005

J Paul Robinson, J Jacobberger

引用次数: 0

Labeling of mitochondria for detection of intercellular mitochondrial transfer. 标记线粒体以检测细胞间线粒体转移。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-06-08 DOI: 10.1016/bs.mcb.2024.05.001

Isamu Taiko, Chika Takano, Shingo Hayashida, Kazunori Kanemaru, Toshio Miki

{"title":"Labeling of mitochondria for detection of intercellular mitochondrial transfer.","authors":"Isamu Taiko, Chika Takano, Shingo Hayashida, Kazunori Kanemaru, Toshio Miki","doi":"10.1016/bs.mcb.2024.05.001","DOIUrl":"10.1016/bs.mcb.2024.05.001","url":null,"abstract":"The phenomenon of intercellular transfer of mitochondria has been reported and has attracted significant interest in recent years. The phenomena involve a range of physiological and pathological conditions, such as tumor growth, immunoregulation, and tissue regeneration. There is speculation on the potential restoration of cellular energy status through the transfer of healthy mitochondria from donor cells to cells with impaired mitochondria. Multiple mechanisms and routes of mitochondria transfer have been suggested, including direct cell-to-cell connections, extracellular vesicles, and cell fusion. However, there is limited understanding regarding the precise mechanisms behind mitochondrial transfer, particularly the initiation signals and the associated processes. In order to explore these fundamental mechanisms of mitochondrial transfer, it is imperative to employ techniques that enable direct labeling of mitochondria. Here, we present a detailed methodology utilizing fluorescent protein tagging to visualize mitochondria. The molecular biological techniques applied in this study entail the precise localization of mitochondria with reduced cytotoxicity. This approach facilitates the direct observation of transferred mitochondria through fluorescent and confocal microscopy. The described method can be readily implemented in other mammalian cell types with few modifications, enabling the continuous monitoring of mitochondrial trafficking processes over an extended period.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"194 ","pages":"1-17"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143586229","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

Quantifying force-mediated antigen extraction in the B cell immune synapse using DNA-based tension sensors. 使用基于dna的张力传感器定量B细胞免疫突触中力介导的抗原提取。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-04-14 DOI: 10.1016/bs.mcb.2024.03.002

Hannah C W McArthur, Anna T Bajur, Katelyn M Spillane

引用次数: 0

Analysis of cytosolic mtDNA release during Staphylococcus aureus infection. 金黄色葡萄球菌感染时胞浆mtDNA释放分析。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-04 DOI: 10.1016/bs.mcb.2024.09.003

Caterina Licini, Gloria D'Achille, Nada Dhaouadi, Ilaria Nunzi, Fabio Marcheggiani, Matteo Fabbri, Monica Mattioli-Belmonte, Gianluca Morroni, Saverio Marchi

引用次数: 0

Human cancer cells xenografts to assess the efficacy of granulysin-based therapeutics. 人类癌细胞异种移植评估颗粒蛋白为基础的治疗效果。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-06-17 DOI: 10.1016/bs.mcb.2024.05.006

Raquel Ibáñez-Pérez, Alberto Anel

{"title":"Human cancer cells xenografts to assess the efficacy of granulysin-based therapeutics.","authors":"Raquel Ibáñez-Pérez, Alberto Anel","doi":"10.1016/bs.mcb.2024.05.006","DOIUrl":"10.1016/bs.mcb.2024.05.006","url":null,"abstract":"9-kDa Granulysin is a protein present in the granules of human activated cytotoxic T lymphocytes and natural killer cells. It has been shown to exert cytolytic activity against a wide variety of microbes: bacteria, fungi, yeast and protozoa. Recombinant isolated granulysin is also capable of inducing tumor cell death, so it could be used as an anti-tumor therapy. Our group has developed granulysin-based immunotoxins in order to target granulysin to tumor cells in vivo. We describe in this chapter the suitable animal model used for testing these immunotoxins against human tumor cells in preclinical assays. This method consists in the xenotransplantation of a given number of human tumor cells subcutaneously in nude mice of the Swiss nu/nu strain or homozygous for the nude gene. Nude mice are immune-deficient, with a very reduced number of T cells, being unable to reject efficiently allo- or xeno-transplanted tissues. Using this approach we follow tumor growth in the different experimental conditions assayed, in the presence or absence of treatment with granulysin alone or with the tumor-directed immunotoxins. We also estimate in this model the possible adverse effects of the treatment in the absence of tumor development. Finally, after sacrifice of the experimentation animals, we use several immunohistochemical assays to study the effect of the treatment on the tumors and the presence of apoptotic cell death markers in the tumor tissue.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"192 ","pages":"83-99"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039853","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

Comprehensive prediction of tumor neoantigens with nextNEOpi. 利用nextNEOpi综合预测肿瘤新抗原。

4区生物学

Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2025-02-06 DOI: 10.1016/bs.mcb.2025.01.007

Markus Ausserhofer, Dietmar Rieder, Francesca Finotello

{"title":"Comprehensive prediction of tumor neoantigens with nextNEOpi.","authors":"Markus Ausserhofer, Dietmar Rieder, Francesca Finotello","doi":"10.1016/bs.mcb.2025.01.007","DOIUrl":"https://doi.org/10.1016/bs.mcb.2025.01.007","url":null,"abstract":"Immunotherapy has revolutionized cancer treatment by harnessing the immune system to target tumor cells expressing neoantigens. Neoantigens are peptides arising from tumor-specific aberrations that are presented by cancer cells and recognized by T cells. The computational prediction of cancer neoantigens from somatic mutations and other tumor-specific aberrations using patients' sequencing data is key for the investigation of anticancer immune responses and for the design of personalized immunotherapies. However, neoantigen prediction requires the implementation of complex computational pipelines to distill large-scale information from RNA and DNA sequencing data and derive neoantigen candidates together with associated features for their prioritization and selection. We previously developed nextNEOpi, a comprehensive and stand-alone bioinformatics pipeline that not only predicts class-I and -II neoantigens and fusion neoantigens, but also sheds light onto the tumor-immune cell interface, quantifying neoantigen clonality, immunogenicity, and tumor-specific metrics like tumor mutational burden and immune-cell receptor repertoire diversity. In this chapter, we showcase the main capabilities of the nextNEOpi pipeline by analyzing genomic and transcriptomic data generated from multiple biopsies collected from patients with lung cancer.","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"196 ","pages":"113-137"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144667964","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