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 ...
We apply this assay to investigate the agonist activity of the T cell co-stimulatory molecule glucocorticoid-induced TNFR-related protein (GITR). We detail a protocol for concurrent assessment of multiple levels of T cell functional modulation upon GITR engagement, including T cell priming, activation and effector function, in a single assay.
Protocol for in vitro isolation, induction, expansion, and ...
2022年12月16日 · Here, we present schemes for the isolation and culture of natural human regulatory T cells (nTregs) and in vitro-induced Tregs (iTregs). Appropriate concentrations of TGF-β, IL-2, retinoic acid (atRA), and rapamycin were used to promote proliferation to meet sample needs in basic research, especially in technologies such as sequencing.
In vitro modeling of CD8 + T cell exhaustion enables CRISPR
2023年8月18日 · In vitro models of T ex are easily customizable and quickly generate high cellular yield, enabling CRISPR screening and other high-throughput assays. We established an in vitro model of chronic stimulation and benchmarked key phenotypic, functional, transcriptional, and epigenetic features against bona fide in vivo T ex.
Targeted in-vitro-stimulation reveals highly proliferative multi …
2019年9月30日 · In-vitro-expansion of T cells is a well-established method, in which lysates of infected cells or viral protein-spanning synthetic peptides can be used for sensitization of T cells against viruses of choice but, beside high costs, it is very time-consuming until a therapeutic product is generated and ready for transfusion.
In Vitro Stimulation and Expansion of Human Tumour‐Reactive …
Here, we investigated the antigen-independent activation and the expansion of human T cells in peripheral blood mononuclear cells (PBMCs) and in tumour-reactive CTLs using Dynabeads coated with monoclonal antibodies to CD3 and to the …
Frontiers | Optimized Protocols for In-Vitro T-Cell-Dependent and …
2022年6月28日 · In this study, we report the optimized and efficient protocols for in-vitro B-cell differentiation using both TD and TI stimulation while requiring very low numbers of B cells.
Optimized Protocols for In-Vitro T-Cell-Dependent and T-Cell ...
2022年6月29日 · Our methodology demonstrates optimized TD and TI stimulation protocols for more in-depth analysis of B-cell differentiation in primary human B-cell and PBMC cultures while requiring low amounts of B cells, making them ideally suited for future clinical and research studies on B-cell differentiation …
Frontiers | Refined analytical pipeline for the pharmacodynamic ...
2024年4月23日 · Current approaches aiming to enrich antigen-specific T cells from human Peripheral Blood Mononuclear Cells (PBMCs) depend on in vitro culturing in presence of their cognate peptides and cytokines. In the present work, we improved a standard, publicly available protocol for T-cell immune analyses based on the in vitro expansion of T cells.
In Vitro Stimulation with Live SARS-CoV-2 Suggests Th17 …
2022年9月16日 · In this study, we investigated the SARS-CoV-2-specific CD4+ T cell phenotype in comparison with H1N1 influenza-specific CD4+ T cells. We determined the levels of SARS-CoV-2- and H1N1-specific CD4+ T cell responses in subjects recovered from COVID-19 one to 15 months ago by stimulating PBMCs with live SARS-CoV-2 or H1N1 influenza viruses.