Flow Cytometry: A Versatile and Powerful Tool for Drug Discovery and Development DOI Creative Commons
Başak Aru, Gülderen Yanıkkaya Demirel

Pharmedicine journal., Год журнала: 2024, Номер 1(1), С. 1 - 19

Опубликована: Фев. 29, 2024

Flow cytometry, a pivotal tool in clinical and research labs since the discovery of cell markers mid-1970s, plays crucial role across all phases drug discovery. Modern flow cytometers can detect rare types relevant to disease pathogenesis, measure numerous parameters simultaneously, thus, offer versatility screening. In studies, cytometry contributes assessment pharmacokinetics, pharmacodynamics safety animal models trials. It also be used monitor efficacy identify biomarkers for diagnosis prognosis. essence, is versatile, instrumental technique that supports from target identification through development, limited only by creativity researcher availability fluorescent labels or specific size/scatter related findings. This review article focuses on use development summarizing not conventional assays such as immunophenotyping, measurement programmed death pathways division provide insights into effects patient responses, but novel approaches including mass spectral droplet cytometry. Keywords: discovery,

Язык: Английский

An autologous ex vivo model for exploring patient-specific responses to viro-immunotherapy in glioblastoma DOI Creative Commons

Εftychia Stavrakaki,

Wouter van den Bossche,

Lisette B. Vogelezang

и другие.

Cell Reports Methods, Год журнала: 2024, Номер 4(3), С. 100716 - 100716

Опубликована: Март 1, 2024

Oncolytic virus (OV) clinical trials have demonstrated remarkable efficacy in subsets of patients with glioblastoma (GBM). However, the lack tools to predict this response hinders advancement a more personalized application OV therapy. In study, we characterize an ex vivo co-culture system designed examine immune infection patient-derived GBM neurospheres presence autologous peripheral blood mononuclear cells (PBMCs). Co-culture conditions were optimized retain viability and functionality both tumor PBMCs, effectively recapitulating well-recognized immunosuppressive effects GBM. Following infection, observed elevated secretion pro-inflammatory cytokines chemokines, including interferon γ, necrosis factor α, CXCL9, CXCL10, marked changes cell activation markers. Importantly, treatment induced unique patient-specific responses. summary, our platform presents avenue for screening viro-immunotherapies GBM, offering promise as potential tool future patient stratification

Язык: Английский

Процитировано

4
Ultrasonic Aspiration-Acquired Glioblastoma Tissue Preserves Lymphocyte Phenotype and Viability, Supporting Its Use for Immunological Studies DOI Open Access

Εftychia Stavrakaki,

Zineb Belcaid, Rutger K. Balvers

и другие.

Cancers, Год журнала: 2025, Номер 17(4), С. 603 - 603

Опубликована: Фев. 11, 2025

Background and Objective: Access to high-quality patient-derived brain tumor tissues is instrumental for translational neuro-oncology research. Glioblastoma material resected by ultrasonic aspiration (UA) during surgery offers an abundant source of material; however, it generally not used research experiments. We hypothesize that UA-derived tissue represents a accurately reflects the immune infiltrates glioblastomas. Methods: In this study, we have utilized performed head-to-head comparison with paired resection from vital core same patient. A combination 16 fluorochrome-conjugated antibodies was designed identify tumor-infiltrating T, B, NK lymphocytes characterize TILs spectral flow cytometry. Furthermore, 5-plex panel spatially T cells, macrophages, cells on UA tissues. Results: UA-obtained exhibited comparable yield viability, as well abundance compared sample-derived cells. Importantly, observed there high concordance respect expression intensities checkpoints in both types samples. Conclusions: These findings underscore feasibility reliability utilizing aspiration-acquired glioblastoma tissue.

Язык: Английский

Процитировано

0
A Tandem Cytometry Platform for Single-Cell Analysis of Protein and Metabolites DOI

Murong Du,

Xingyu Pan,

Yamei Peng

и другие.

Analytical Chemistry, Год журнала: 2025, Номер unknown

Опубликована: Март 25, 2025

Multiomics analysis at the single-cell level is essential for both fundamental research and clinical applications, with proteomics metabolomics being particularly crucial providing insights into cellular states functions. The state of art flow cytometry has shown great potential in identifying proteins, while emerging metabolite mass spectrometry techniques address detection. Herein, we propose a tandem platform that integrates fluorescence electrospray ionization one-step protein metabolites. An algorithm was established to correlate multidimensional information individual cells, additional data processing modules designed ensure accuracy facilitate further analysis. demonstrated efficacy profiling breast cancer under hypoxic conditions, revealing metabolic shifts decreased glutathione increased l-glutamine levels, indicative hypoxia-inducible factor activity. This introduces powerful analytical capability promises elevate precision cell-based diagnostics therapeutic strategies.

Язык: Английский

Процитировано

0
Characterization of Human B Cell Hematological Malignancies Using Protein-Based Approaches DOI Open Access
Cristina Jiménez, Alba Garrote-de-Barros, Carlos López-Portugués

и другие.

International Journal of Molecular Sciences, Год журнала: 2024, Номер 25(9), С. 4644 - 4644

Опубликована: Апрель 24, 2024

The maturation of B cells is a complex, multi-step process. During cell differentiation, errors can occur, leading to the emergence aberrant versions that, finally, constitute malignant tumor. These malignancies are classified into three main groups: leukemias, myelomas, and lymphomas, latter being most heterogeneous type. Since their discovery, multiple biological studies have been performed characterize these diseases, aiming define specific features determine potential biomarkers for diagnosis, stratification, prognosis. rise advanced -omics approaches has significantly contributed this end. Notably, proteomics strategies appear as promising tools comprehensively profile final molecular effector cells. In narrative review, we first introduce together with relevant approaches. Then, describe core conducted in field findings and, evaluate advantages drawbacks flow cytometry, mass spectrometry profiling human disorders.

Язык: Английский

Процитировано

3
Recent Developments in Blood Biomarkers in Neuro-oncology DOI
Jessica Foster, Mateusz Koptyra, Stephen Bagley

и другие.

Current Neurology and Neuroscience Reports, Год журнала: 2023, Номер 23(12), С. 857 - 867

Опубликована: Ноя. 9, 2023

Язык: Английский

Процитировано

4
Development of a Spectral Flow Cytometry Analysis Pipeline for High-Dimensional Immune Cell Characterization DOI Creative Commons

Donald Vardaman,

Md Akkas Ali,

Chase Bolding

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Июнь 22, 2024

Flow cytometry is a widely used technique for immune cell analysis, offering insights into composition and function. Spectral flow allows high-dimensional analysis of cells, overcoming limitations conventional cytometry. However, analyzing data from large antibody panels can be challenging using traditional bi-axial gating strategies. Here, we present novel pipeline designed to improve spectral We employ this method identify rare T populations in aging. isolated splenocytes young (2-3 months) aged (18-19 female mice then stained these with panel 20 fluorescently labeled antibodies. was performed, followed by processing Python within Jupyter Notebook environment perform batch correction, unsupervised clustering, dimensionality reduction, differential expression analysis. Our 3,776,804 cells 11 spleens revealed 34 distinct clusters identified surface marker expression. observed significant differences between mice, certain enriched one age group over the other. Naïve, effector memory, central memory CD8

Язык: Английский

Процитировано

1
42-parameter mass cytometry panel to assess cellular and functional phenotypes of leukocytes in bronchoalveolar lavage of Rhesus macaque DOI Creative Commons
Mohau S. Makatsa, Anna Kus, Alice Wiedeman

и другие.

bioRxiv (Cold Spring Harbor Laboratory), Год журнала: 2024, Номер unknown

Опубликована: Сен. 24, 2024

ABSTRACT This Optimized Multiparameter Immunofluorescence Panel (OMIP) reports on the development of a mass cytometry panel for broad immunophenotyping leukocytes from bronchoalveolar lavage rhesus macaques. Using this panel, we were able to identify myeloid populations such as macrophages, neutrophils, monocytes, and plasmacytoid DCs, basophils lymphoid cell lineages including B cells, natural killer (NK) mucosal associated invariant T (MAIT) γδ CD4 CD8□β CD8 □□ innate cells (ILCs). We also included markers defining memory, differentiation (CCR7, CD28, CD45RA), homing potential (CXCR3), cytotoxic (perforin, granzyme B, K), activation/differentiation (HLA-DR, CD69, IgD) effector function (CD154, IFN-γ, TNF, IL-2, IL-17A, IL-6, IL-1β, CCL4 CD107a). was optimized cryopreserved, splenocytes collected The antibodies selected in are human-specific that have been shown cross-react with non-human primates except CD45 clone D058-1283 which is specific primates.

Язык: Английский

Процитировано

1
Automated EuroFlow approach for standardized in-depth dissection of human circulating B-cells and plasma cells DOI Creative Commons
Alejandro Hernández-Delgado,

Rafael Fluxá,

Martín Pérez‐Andres

и другие.

Frontiers in Immunology, Год журнала: 2023, Номер 14

Опубликована: Окт. 17, 2023

Multiparameter flow cytometry (FC) immunophenotyping is a key tool for detailed identification and characterization of human blood leucocytes, including B-lymphocytes plasma cells (PC). However, currently used conventional data analysis strategies require extensive expertise, are time consuming, show limited reproducibility.Here, we designed, constructed validated an automated database-guided gating (AGI) approach fast standardized in-depth dissection B-lymphocyte PC populations in blood.For this purpose, 213 FC standard (FCS) datafiles corresponding to umbilical cord peripheral samples from healthy patient volunteers, stained with the 14-color 18-antibody EuroFlow BIgH-IMM panel, were used.The antibody panel allowed 117 different subsets. Samples 36 donors 14 that fulfilled strict inclusion criteria analysed by expert cytometrist build database. Data contained was then merged into reference database uploaded Infinicyt software (Cytognos, Salamanca, Spain). Subsequently, compared results manual (MG) performance two classification algorithms -hierarchical algorithm vs two-step algorithm- AGI cell present 5 randomly selected FCS datafiles. The hierarchical showed higher correlation values MG (r2 0.94 vs. 0.88 algorithm) further set 177 against expert-based MG. For virtually all identifiable highly significant observed between approaches (r2>0.81 79% B-cell identified), significantly lower median per sample (6 40 min, p=0.001) MG, respectively both intra-sample (median CV 1.7% 10.4% p<0.001) inter-expert 3.9% 17.3% 2 experts, variability.Our strategies, here proposed faster, more robust, reproducible, subsets circulating blood.

Язык: Английский

Процитировано

3
Altered immune cell phenotypes within chronically ischemic human kidneys distal to occlusive renal artery disease DOI
Xiangyang Zhu, Nattawat Klomjit, Aditya S. Pawar

и другие.

AJP Renal Physiology, Год журнала: 2023, Номер 326(2), С. F257 - F264

Опубликована: Ноя. 30, 2023

Renal artery stenosis (RAS) is a major cause of ischemic kidney disease, which largely mediated by inflammation. Mapping the immune cell composition in kidneys might provide useful insight into disease pathogenesis and uncover therapeutic targets. We used mass cytometry (CyTOF) to explore single-cell unique data set human nephrectomized due chronic occlusive vascular (RAS,

Язык: Английский

Процитировано

3
Comparative assessment of cytometry by time-of-flight and full spectral flow cytometry based on a 33-color antibody panel DOI Creative Commons

Antonia Schäfer,

Sènan Mickaël D'Almeida, Julien Dorier

и другие.

Journal of Immunological Methods, Год журнала: 2024, Номер 527, С. 113641 - 113641

Опубликована: Фев. 15, 2024

Mass cytometry and full spectrum flow have recently emerged as new promising single cell proteomic analysis tools that can be exploited to decipher the extensive diversity of immune repertoires their implication in human diseases. In this study, we evaluated performance mass against using an identical 33-color antibody panel on four healthy individuals. Our data revealed overall high concordance quantification major populations between two platforms a semi-automated clustering approach. We further showed strong correlation cluster assignment when comparing manual automated clustering. Both comparisons minor disagreements rare subpopulations. study both technologies generate highly overlapping results substantiate choice technology is not primary factor for successful biological assessment profiles but must considered broader design framework clinical studies.

Язык: Английский

Процитировано

0