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Evaluating amyloid-beta aggregation and toxicity in transgenic Caenorhabditis elegans models of Alzheimer's disease.
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
{"title":"Evaluating amyloid-beta aggregation and toxicity in transgenic Caenorhabditis elegans models of Alzheimer's disease.","authors":"Leticia Priscilla Arantes, Larissa Marafiga Cordeiro, Félix Alexandre Antunes Soares","doi":"10.1016/bs.mcb.2024.06.001","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.06.001","url":null,"abstract":"<p><p>Alzheimer's disease (AD) is the leading cause of dementia in the elderly, clinically characterized by memory loss, cognitive decline, and behavioral disturbances. Its pathogenesis is not fully comprehended but involves intracellular depositions of amyloid beta peptide (Aβ) and neurofibrillary tangles of hyperphosphorylated tau. Currently, pharmacological interventions solely slow the progression of symptoms. Caenorhabditis elegans (C. elegans) is a simple and valuable organism to study the dynamics of Aβ. It may contribute to advancing our comprehension of AD development and progression, as well as to discovering new treatments. Herein, we describe usual protocols for evaluating Aβ aggregation and toxicity in transgenic C. elegans models of AD (CL2006, CL4176, GMC101, and CL2355 strains) through the visualization and quantification of the peptide with specific fluorescent dyes, in addition to the analysis of particular behaviors (paralysis and chemotaxis associated with learning).</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"192 ","pages":"189-202"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039849","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 cancer cells xenografts to assess the efficacy of granulysin-based therapeutics.
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":"https://doi.org/10.1016/bs.mcb.2024.05.006","url":null,"abstract":"<p><p>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.</p>","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
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
{"title":"Splenocytes antitumor cytotoxicity assessment after prophylactic vaccination or drug treatment of tumor-bearing mice.","authors":"Kenny Misael Calvillo-Rodriguez, Ana Luisa Rivera-Lazarin, Reyes Tamez-Guerra, Ana Carolina Martinez-Torres, Cristina Rodriguez-Padilla","doi":"10.1016/bs.mcb.2024.10.011","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.10.011","url":null,"abstract":"","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"197-210"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008178","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.
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
{"title":"Quantifying force-mediated antigen extraction in the B cell immune synapse using DNA-based tension sensors.","authors":"Hannah C W McArthur, Anna T Bajur, Katelyn M Spillane","doi":"10.1016/bs.mcb.2024.03.002","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.03.002","url":null,"abstract":"<p><p>B cells exert pulling forces against antigen-presenting cells (APCs) to extract antigens for internalization. The application of tugging forces on B cell receptor (BCR)-antigen bonds promotes discrimination of antigen affinities and sensing of APC physical properties. Here, we describe a protocol for preparing antigen-functionalized DNA tension sensors for quantifying force-mediated antigen extraction in the B cell immune synapse. We describe how to attach the sensors to planar lipid bilayers, quantify their surface density, use them to stimulate B cell activation, and analyze the efficiency of antigen extraction in fixed cells by fluorescence microscopy and image analysis. These techniques should be broadly applicable to studies of force-mediated transfer of molecules in cell-cell contacts.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"193 ","pages":"99-126"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143370864","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
Expression, purification and characterization of phosphatidylserine-targeting antibodies for biochemical and therapeutic applications. 用于生化和治疗的磷脂酰丝氨酸靶向抗体的表达、纯化和鉴定。
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-21 DOI: 10.1016/bs.mcb.2024.10.003
Varsha Gadiyar, David C Calianese, Rachael Pulica, Christopher Varsanyi, Ziren Wang, Ahmed Aquib, Alok Choudhary, Raymond B Birge
{"title":"Expression, purification and characterization of phosphatidylserine-targeting antibodies for biochemical and therapeutic applications.","authors":"Varsha Gadiyar, David C Calianese, Rachael Pulica, Christopher Varsanyi, Ziren Wang, Ahmed Aquib, Alok Choudhary, Raymond B Birge","doi":"10.1016/bs.mcb.2024.10.003","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.10.003","url":null,"abstract":"<p><p>The externalization of Phosphatidylserine (PS) from the inner surface of the plasma membrane to the outer surface of the plasma membrane is an emblematic event during apoptosis and serves as a potent \"eat-me\" signal for the efferocytosis of apoptotic cells. Although less well understood, PS is also externalized on live cells in the tumor microenvironment and on live virus-infected cells whereby it serves as an immune modulatory signal that drives tolerance and immune escape. Given the importance of PS in cancer immunology and immune escape, PS-targeting monoclonal antibodies have been characterized with promising immunotherapeutic potential. Here, we describe the cloning and characterization of a series of PS targeting antibodies and their potential use and utility in immuno-oncology.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"15-40"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008118","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
A mouse model to assess immunotherapy-related colitis.
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-04-18 DOI: 10.1016/bs.mcb.2024.03.010
Yaiza Senent, Ana Remírez, Beatriz Tavira, Daniel Ajona
{"title":"A mouse model to assess immunotherapy-related colitis.","authors":"Yaiza Senent, Ana Remírez, Beatriz Tavira, Daniel Ajona","doi":"10.1016/bs.mcb.2024.03.010","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.03.010","url":null,"abstract":"<p><p>Combined blockade of the immune checkpoints PD-1 and CTLA-4 has shown remarkable efficacy in patients with melanoma, renal cell carcinoma, non-small-cell lung cancer and mesothelioma, among other tumor types. However, a proportion of patients suffer from serious immune-related adverse events (irAEs). In severe cases, a reduction of the doses or the complete cessation of the treatment is required, limiting the antitumor efficacy of these treatments. Colitis is among the most frequent and problematic irAE associated with immune checkpoint blockade. In this context, animal models that recapitulate the pathophysiological features of immunotherapy-related colitis are needed. In this manuscript, we describe our experience with a mouse model in which the combined CTLA-4 and PD-1 blockade exacerbates the deleterious effects of dextran sulfate sodium (DSS)-induced colitis. This model may constitute a valuable tool for the study of immunotherapy-related colitis.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"192 ","pages":"33-38"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039665","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
Development of an in vivo ovarian cancer peritoneal carcinomatosis model for radioimmunotherapy testing.
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-06-17 DOI: 10.1016/bs.mcb.2024.05.013
Clara Diaz Garcia-Prada, Salima Atis, Jean-Pierre Pouget, Julie Constanzo
{"title":"Development of an in vivo ovarian cancer peritoneal carcinomatosis model for radioimmunotherapy testing.","authors":"Clara Diaz Garcia-Prada, Salima Atis, Jean-Pierre Pouget, Julie Constanzo","doi":"10.1016/bs.mcb.2024.05.013","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.05.013","url":null,"abstract":"<p><p>Currently, Ovarian Cancer (OC) is the most lethal gynecological malignancy. In most patients, it progresses without clinical signs or symptoms, leading to a late diagnosis when it has already spread in the peritoneal cavity as peritoneal carcinomatosis (PC). To date, OC PC management is based on cytoreductive surgery to remove the macroscopic disease, followed by chemotherapy. Many patients respond to this treatment, but disease recurs in 70-90% of them. Therefore, new therapeutic approaches are needed. The field of targeted radionuclide therapy (TRT) has witnessed considerable progress and several radiopharmaceuticals have been approved in the last decade. In TRT, radiolabeled molecules are injected to specifically recognize, irradiate, and kill tumor cells. TRT is a multisite radiotherapy that delivers dose to all malignant lesions. Therefore, TRT could be an alternative approach for OC PC because conventional external beam radiotherapy cannot be used at curative dose due to toxicity to healthy tissues. Here, we describe an OC PC model based on grafting human SK-OV-3 OC cells in the peritoneal cavity of immunodeficient mice. We also explain how to label trastuzumab with lutetium-177 to specifically target and irradiate SK-OV-3 cell nodules in these mice, and how to monitor the response to this TRT in vivo. With minor variations, the same technique can be conveniently applied to a variety of human (or mouse) tumors.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"192 ","pages":"131-157"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039848","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
The current models unravel the molecular mechanisms underlying the intricate pathophysiology of Alzheimer's disease using zebrafish.
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-04-18 DOI: 10.1016/bs.mcb.2024.03.009
Baban Thawkar, Ginpreet Kaur
{"title":"The current models unravel the molecular mechanisms underlying the intricate pathophysiology of Alzheimer's disease using zebrafish.","authors":"Baban Thawkar, Ginpreet Kaur","doi":"10.1016/bs.mcb.2024.03.009","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.03.009","url":null,"abstract":"<p><p>The foremost cause of dementia is Alzheimer's disease (AD). The vital pathological hallmarks of AD are amyloid beta (Aβ) peptide and hyperphosphorylated tau (p-tau) protein. The current animal models used in AD research do not precisely replicate disease pathophysiology, making it difficult for researchers to quickly and effectively gather data or screen potential therapy possibilities. Several transgenic animals are used as models for AD; however, they have cost and time concerns. Zebrafish (Danio rerio) has become a suitable model organism for high-throughput pharmacological screening of neuroactive substances and neurodegenerative research. The past few decades have seen a significant increase in research on AD. The fight against amyloidosis has, however, been unexpectedly unsuccessful. It may be due to a need for more relevant in vivo models for high throughput screening, which emphasizes the need to find other anti-AD models. Alternative animal models, including zebrafish, have developed into a potentially useful research tool that must be employed for AD research to be effective. Only a few comprehensive zebrafish models exhibiting AD-like pathogenesis have been reported in the literature, and this book chapter describes these models.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"192 ","pages":"17-31"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143039913","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
Evaluation of lymphocyte infiltration into cancer spheroids by immunofluorescent staining and 3D imaging. 免疫荧光染色和三维成像评价淋巴细胞浸润癌球体。
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-14 DOI: 10.1016/bs.mcb.2024.10.015
Mireia Cruz De Los Santos, Andreas Lundqvist
{"title":"Evaluation of lymphocyte infiltration into cancer spheroids by immunofluorescent staining and 3D imaging.","authors":"Mireia Cruz De Los Santos, Andreas Lundqvist","doi":"10.1016/bs.mcb.2024.10.015","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.10.015","url":null,"abstract":"<p><p>In recent years, three-dimensional (3D) cultures of tumor cells has emerged as an important tool in cancer research. The significance of 3D cultures, such as tumor spheroids, lies in their ability to mimic the in vivo tumor microenvironment more precisely, offering a nuanced understanding of immune responses within the context of tumor progression. In fact, the infiltration of cytotoxic lymphocytes is key to determining patients' prognosis in several types of cancer and response to immunotherapy. Therefore, harnessing the cytotoxic and infiltration potential of immune cells is a promising avenue for developing effective therapies. This protocol offers a straightforward approach for analyzing infiltrating lymphocytes in tumor spheroids by confocal microscopy imaging. Although it specifically involves utilizing tumor spheroids and culture with autologous tumor-infiltrating T lymphocytes (TILs), the protocol can be adapted for other immune cell types, such as NK cells.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"269-287"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008111","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
Ex vivo assessment of human neutrophil motility and migration. 人中性粒细胞运动和迁移的离体评估。
4区 生物学
Methods in cell biology Pub Date : 2025-01-01 Epub Date: 2024-11-19 DOI: 10.1016/bs.mcb.2024.10.008
Noor A M Bakker, Claudia Burrello, Karin E de Visser
{"title":"Ex vivo assessment of human neutrophil motility and migration.","authors":"Noor A M Bakker, Claudia Burrello, Karin E de Visser","doi":"10.1016/bs.mcb.2024.10.008","DOIUrl":"https://doi.org/10.1016/bs.mcb.2024.10.008","url":null,"abstract":"<p><p>Neutrophils are pivotal in orchestrating tumor-induced systemic inflammation and are increasingly recognized for their critical involvement in both the initiation and progression of cancer. A fundamental facet of neutrophil biology is their migratory capacity, which enables them to extravasate and infiltrate tumors in other tissues, where they carry out essential effector functions. Unraveling the intricate mechanisms of neutrophil motility and migration is crucial for comprehending immune responses and inflammatory processes, shedding light on their substantial contribution to cancer progression. Here, we provide a comprehensive protocol to assess direct ex vivo motility and migration of freshly isolated human neutrophils, offering valuable insights into their behavior.</p>","PeriodicalId":18437,"journal":{"name":"Methods in cell biology","volume":"191 ","pages":"115-133"},"PeriodicalIF":0.0,"publicationDate":"2025-01-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143008115","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
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