Easily and efficiently isolate highly purified human naïve CD8+ T cells (CD8+CD45RA+CCR7+ and CD45RO-CD57-CD56-) from fresh human peripheral blood mononuclear cell (PBMC) samples by immunomagnetic negative selection, with the EasySep™ Human Naïve CD8+ T Cell Isolation Kit. Widely used in published research for more than 20 years, EasySep™ combines the specificity of monoclonal antibodies with the simplicity of a column-free magnetic system.
In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD4, CD14, CD16, CD19, CD20, CD36, CD45RO, CD56, CD57, CD94, CD123, CD244, TCRγ/δ, and glycophorin A. The magnetically labeled cells are then separated from the untouched desired naïve CD8+ T cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired naïve CD8+ T cells are ready for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.
For even faster cell isolations, we recommend the EasySep™ Human Naïve CD8+ T Cell Isolation Kit II (17968) which isolates cells in as little as 11 minutes.
Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
In this EasySep™ negative selection procedure, unwanted cells are labeled with antibody complexes and magnetic particles. Unwanted cells expressing the following markers are targeted for removal: CD4, CD14, CD16, CD19, CD20, CD36, CD45RO, CD56, CD57, CD94, CD123, CD244, TCRγ/δ, and glycophorin A. The magnetically labeled cells are then separated from the untouched desired naïve CD8+ T cells by using an EasySep™ magnet and simply pouring or pipetting the desired cells into a new tube. Following magnetic cell isolation, the desired naïve CD8+ T cells are ready for downstream applications such as flow cytometry, culture, or DNA/RNA extraction.
For even faster cell isolations, we recommend the EasySep™ Human Naïve CD8+ T Cell Isolation Kit II (17968) which isolates cells in as little as 11 minutes.
Learn more about how immunomagnetic EasySep™ technology works or how to fully automate immunomagnetic cell isolation with RoboSep™. Explore additional products optimized for your workflow, including culture media, supplements, antibodies, and more.
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Protocols and Documentation
Find supporting information and directions for use in the Product Information Sheet or explore additional protocols below.
Applications
This product is designed for use in the following research area(s) as part of the highlighted workflow stage(s). Explore these workflows to learn more about the other products we offer to support each research area.
Resources and Publications
Educational Materials (2)
Publications (4)
Comparative analysis of the DNA methylation landscape in CD4, CD8, and B memory lineages. Clinical epigenetics 2022 dec
Abstract
BACKGROUND There is considerable evidence that epigenetic mechanisms and DNA methylation are critical drivers of immune cell lineage differentiation and activation. However, there has been limited coordinated investigation of common epigenetic pathways among cell lineages. Further, it remains unclear if long-lived memory cell subtypes differentiate distinctly by cell lineages. RESULTS We used the Illumina EPIC array to investigate the consistency of DNA methylation in B cell, CD4 T, and CD8 T na{\{i}}ve and memory cells states. In the process of na{\"{i}}ve to memory activation across the three lineages we identify considerable shared epigenetic regulation at the DNA level for immune memory generation. Further in central to effector memory differentiation our analyses revealed specific CpG dinucleotides and genes in CD4 T and CD8 T cells with DNA methylation changes. Finally we identified unique DNA methylation patterns in terminally differentiated effector memory (TEMRA) CD8 T cells compared to other CD8 T memory cell subtypes. CONCLUSIONS Our data suggest that epigenetic alterations are widespread and essential in generating human lymphocyte memory. Unique profiles are involved in methylation changes that accompany memory genesis in the three subtypes of lymphocytes."
IL-9/STAT3/fatty acid oxidation-mediated lipid peroxidation contributes to Tc9 cell longevity and enhanced antitumor activity. The Journal of clinical investigation 2022 apr
Abstract
CD8+ T cell longevity regulated by metabolic activity plays important roles in cancer immunotherapy. Although in vitro-polarized, transferred IL-9-secreting CD8+ Tc9 (cytotoxic T lymphocyte subset 9) cells exert greater persistence and antitumor efficacy than Tc1 cells, the underlying mechanism remains unclear. Here, we show that tumor-infiltrating Tc9 cells display significantly lower lipid peroxidation than Tc1 cells in several mouse models, which is strongly correlated with their persistence. Using RNA-sequence and functional validation, we found that Tc9 cells exhibited unique lipid metabolic programs. Tc9 cell-derived IL-9 activated STAT3, upregulated fatty acid oxidation and mitochondrial activity, and rendered Tc9 cells with reduced lipid peroxidation and resistance to tumor- or ROS-induced ferroptosis in the tumor microenvironment. IL-9 signaling deficiency, inhibiting STAT3, or fatty acid oxidation increased lipid peroxidation and ferroptosis of Tc9 cells, resulting in impaired longevity and antitumor ability. Similarly, human Tc9 cells also exhibited lower lipid peroxidation than Tc1 cells and tumor-infiltrating CD8+ T cells expressed lower IL9 and higher lipid peroxidation- and ferroptosis-related genes than circulating CD8+ T cells in patients with melanoma. This study indicates that lipid peroxidation regulates Tc9 cell longevity and antitumor effects via the IL-9/STAT3/fatty acid oxidation pathway and regulating T cell lipid peroxidation can be used to enhance T cell-based immunotherapy in human cancer.
Age-Associated Changes to Lymph Node Fibroblastic Reticular Cells. Frontiers in aging 2022
Abstract
The decreased proportion of antigen-inexperienced, na{\{i}}ve T cells is a hallmark of aging in both humans and mice and contributes to reduced immune responses particularly against novel and re-emerging pathogens. Na{\"{i}}ve T cells depend on survival signals received during their circulation among the lymph nodes by direct contacts with stroma in particular fibroblastic reticular cells. Macroscopic changes to the architecture of the lymph nodes have been described but it is unclear how lymph node stroma are altered with age and whether these changes contribute to reduced na{\"{i}}ve T cell maintenance. Here using 2-photon microscopy we determined that the aged lymph node displayed increased fibrosis and correspondingly that na{\"{i}}ve T-cell motility was impaired in the aged lymph node especially in proximity to fibrotic deposition. Functionally adoptively transferred young na{\"{i}}ve T-cells exhibited reduced homeostatic turnover in aged hosts supporting the role of T cell-extrinsic mechanisms that regulate their survival. Further we determined that early development of resident fibroblastic reticular cells was impaired which may correlate to the declining levels of na{\"{i}}ve T-cell homeostatic factors observed in aged lymph nodes. Thus our study addresses the controversy as to whether aging impacts the composition lymph node stroma and supports a model in which impaired differentiation of lymph node fibroblasts and increased fibrosis inhibits the interactions necessary for na{\"{i}}ve T cell homeostasis."
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