One of the most common ways to assess T cell activation is to measure T cell proliferation upon in vitro stimulation of T cells via antigen or agonistic antibodies to TCR. This protocol is written as a starting point for examining in vitro proliferation of mouse splenic T-cells and human peripheral T cells stimulated via CD3.
In vitro assays for effector T cell functions and activity of ...
In this chapter, we describe an optimized version of T cell suppression assay designed to test the capacity of immune co-stimulatory agents to enhance priming and activation of T cells in the presence of Tregs.
Protocol for the assessment of human T cell activation by real …
Mar 18, 2022 · Here, we describe a simple and efficient protocol for isolating human peripheral blood mononuclear cells (PBMCs) and T cells, and the subsequent assessment of T cell glycolysis using Seahorse analyzer.
T Cell Activation Bioassays - Promega Corporation
Our ICH-prequalified bioassays enable simple, scalable solutions for drug screening and development. These assays consist of a genetically engineered T cell line that expresses a luciferase reporter driven by either an NFAT response element or an IL-2 promoter.
T-cell activation-induced marker assays in health and disease
Activation-induced marker (AIM) assays have proven to be an accessible and rapid means of antigen-specific T-cell detection. The method typically involves short-term incubation of whole blood or peripheral blood mononuclear cells with antigens of interest, where autologous antigen-presenting cells p …
T Cell Assays - Reaction Biology
We have developed a systemic approach to T cell activation for drug discovery. T-cell activation can be checked by the expression of a luciferase reporter driven by either an NFAT response element or an IL-2 promoter. Both cell lines are available for assessing T cell activation by employing bioluminescent methods or ELISA kits. Example data.
The T Cell Activation Bioassay reflects the mechanism of action (MOA) of biologics designed to engage the TCR and induce TCR-mediated T cell activation. Specifically, anti-CD3 antibody-mediated luminescence is detected using either the
T Cell Activation Assay Service - Horizon Discovery
The T cell activation assay is a rapid in-vitro monoculture system to assess the desired effect of your developed molecule to promote or block cell proliferation and activation in CD4+ and CD8+T cell subtypes.
T Cell Activation Bioassay (NFAT/IL-2) - Promega Corporation
When the TCR/CD3 Effector Cells (NFAT) are engaged with an appropriate TCR/CD3 ligand or anti-TCR/CD3 antibody, the TCR transduces intracellular signals resulting in NFAT-RE-mediated luminescence.
T-cell activation–induced marker assays in health and disease
Feb 24, 2023 · Recognition of peptide–MHC complexes by T-cell receptors then induces upregulation of activation markers on the T cells that can be detected by flow cytometry. In this review, we highlight the most widely used activation markers for assays in the literature while identifying nuances and potential downfalls associated with the technique.
In-depth characterization of T cell responses with a combined ...
Dec 9, 2024 · Upon T cell activation, the expression of 4-1BB, CD25, and CD69 increases and peaks at 24-, 48-, and 16-h post-stimulation, respectively. ... This protocol was developed in order to address the need for an in-depth T cell characterization assay that could simultaneously determine (i) the type of T cell responding, (ii) the activation of the T ...
EpiScreen® 2.0 Time Course T Cell Assay - Abzena
By measuring CD4+ T cell responses ex vivo, the assay assesses the strength and kinetics of an individual’s T cell response to a protein therapeutic, thereby giving an indication of an immunogenic response against the protein.
Assessing T-Cell Assays and Immune Response Sensitivity
Oct 7, 2024 · T-cell activation assays are instrumental in dissecting the complexities of immune responses. These assays are designed to measure the activation and proliferation of T-cells when exposed to specific stimuli.
Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation ...
Jan 1, 2021 · Here we describe an NFAT-driven fluorescent reporter system and T-cell multiplexing strategy for large-scale screening of TCR and peptide-MHC complex interactions, to surmount this limiting step in determination of TCR antigen specificity. Figure 1. Principle of the assay – strategy of reporter T-cell generation.
Dual Activation‐Induced Marker Combinations Efficiently Identify …
Individual AIMs detect activated CD4 + T cells with good sensitivity and high specificity but poorly distinguish true-positive and false-positive events. (A) Schematic representation of the experimental strategy, created with the help of Biorender.com. (B) CTV staining of CD4 + and CD8 + T cells stimulated with plate-bound α-CD3 + α-CD28 antibodies or left unstimulated.
T Cell Activation Bioassay (IL-2) Protocol - Promega Corporation
The T Cell Activation Bioassay (IL-2), is a bioluminescent cell-based assay that overcomes the limitations of existing assays and can be used for the discovery and development of novel biologic and cell therapies aimed at inducing, strengthening and/or engineering T cell responses.
T Cell Activation and Proliferation Assays - ProMab
ProMab has developed a systematic approach to T cell activation and proliferation assays for IO products discovery. The nuclear factor of activated T cells (NFAT), IL-2 or other kinds of factors will be detected by bioluminescent methods or ELISA kits.
T-cell activation - Takara Bio
To demonstrate the utility of GoStix Plus assays for rapid quantitation of cytokines in immune cell culture applications, primary human T cells were isolated from peripheral blood mononuclear cells (PBMCs) and activated using either of three different methods.
Multiplex T-cell Stimulation Assay Utilizing a T-cell Activation ...
Jan 20, 2021 · Next-generation sequencing methods allow for the rapid identification of thousands of TCR clonotypes from single T-cells, and thus there is a need to determine cognate antigens for identified TCRs.
Multiplex T Cell Stimulation Assay Utilizing a T Cell Activation ...
Apr 9, 2020 · Using a NFAT-based fluorescent reporter system, we generated T-hybridoma cell lines that express intensely fluorescent proteins in response to antigen stimulation and constitutively express additional fluorescent proteins, which serve as identifiers of each T-hybridoma expressing a unique TCR.
T Cell Types, Activation, Polarization & Function - Assay Genie
T-cells, vital components of the immune system, encompass various functions and types, including cytotoxic T cells, helper T cells, and regulatory T cells. Their activation and polarization play critical roles in immune responses.
In vitro assays for effector T cell functions and activity of ...
Jan 1, 2020 · Here, we describe an optimized version of T cell suppression assay designed to test the effect of immunomodulatory agents on T cell function and activation. We apply this assay to investigate the agonist activity of the T cell co-stimulatory molecule glucocorticoid-induced TNFR-related protein (GITR).
Hypoxia-induced autophagy in pancreatic cancer counteracts the ...
5 days ago · The content of cytokines and the activation level of CD8 + T cells were measured by enzyme-linked immunosorbent assay (ELISA) and flow cytometry. MHC-I expression in PC cells (membranes) was ...
CD69+CD103+CD8+ tissue-resident memory T cells possess …
Dec 18, 2024 · In vitro CD8 + T cell activation and inhibition assay. Peripheral blood mononuclear cells (PBMCs) were isolated by Ficoll (Lymphoprep, AS1114546, Axis-Shield) density gradient centrifugation method , and CD8 + T cells were sorted using …
C3+ cancer-associated fibroblasts promote tumor growth and …
Dec 20, 2024 · Background Gastric cancer (GC) remains one of the most lethal malignancies globally, with limited therapeutic options. Cancer-associated fibroblasts (CAFs), a diverse population of stromal cells within the tumor microenvironment (TME), play a central role in tumor progression and therapeutic resistance. However, the specific markers identifying tumor …
Characterization of anti-drug antibody responses to the T-cell …
May 31, 2024 · Here, the PK assay of a T-cell-engaging bispecific antibody, cibisatamab, was developed based on its mechanism of action (MoA). Using critical monoclonal anti-idiotypic (anti-ID) antibody positive controls as ADA surrogates, the impact on …
Dynamic control of RNA-DNA hybrid formation orchestrates ... - Cell …
Dec 19, 2024 · Song et al. demonstrate that the dynamic regulation of RNA-DNA hybrid formation at stalled forks by RNA polymerase II and a newly identified RNA-DNA resolver, DDX39A, precisely orchestrates the timing of DNA2 nuclease activation. This regulation is essential for stalled fork protection, processing, and restart, ultimately safeguarding genome stability.