Whether immuno-oncology research, basic drug discovery research, advanced-stage clinical studies, or disease diagnostic phenotyping, immune cells are routinely analyzed. Flow cytometry has become the technology of choice for this since it provides the possibility of analyzing a large number of parameters simultaneously, quickly, and for a reasonable cost.
Immuno-oncology research focuses heavily on immune cells implicated in the tolerance or rejection of cancer. In a cancer immunity cycle, cancer cells release inflammation markers that attract the attention of antigen-presenting cells, such as NK cells, dendritic cells, and macrophages, which in turn activate T cells. These activated T cells infiltrate the tumor, recognize tumor-specific antigens, and begin the process of killing tumor cells. Miltenyi Biotec offers pre-tested antibody panels for phenotyping of immune cells involved in the tumorigenesis process.
Flow cytometry allows identification of cellular markers through surface or intracellular staining techniques. These markers can be used to identify different cell types, which is useful in developing cell-specific therapies.
Furthermore, flow cytometry’s ability to characterize, count, and sort fluorescently labeled cells based on their phenotype affords it yet another advantage in immuno-oncological applications. Quantification and characterization of specific subsets of cell populations allows us to better ascertain the immunogenicity of a therapy, and thereby enhance our understanding of complex immune responses.
Our new and advanced flow cytometry instruments, as well as an expanded range of antibodies and fluorochromes to accompany the technique, enable more accurate phenotyping of cells, which in turn enables enhanced identification of cellular distribution in the cancer environment.
Key features and advantages of flow cytometry include:
Designing and testing flow cytometry antibody panels can be laborious, with reproducible results often a challenge. Learn more about an easier and more reproducible method that: Eliminates assay design steps, reduces hands-on time at the flow cytometer and removes manual gating steps.
In this table you can find a common immunophenotyping markers to distinguish different cell types in immuno-oncology research. Click on the respective marker in order to find relevant antibodies for detection.
|Cell type||Surface |
|Secreted proteins||Activation markers||Exhaustion markers||Proliferation markers||Apoptosis markers||Cell signaling markers|
|T cells||CD3, CD4, CD8, CD27, CD45RA, CD62L||T-bet, GATA3, FoxP3, RORγt||IFNγ, IL-2, TNFα, IL-4, IL-5, IL-17, IL-10||CD25, CD69, CD95, CD134, CD137, CD154||PD-1, CTLA-4, LAG-3, TIM-3, TIGIT, CD244, CD272, CD278||Ki-67, PCNA, Bcl-6||pH2A.X, Caspase, Bcl-2||p-Lck, p-ZAP70, p-PLCy2, p-ERK, p-NFκB, p-Akt, p-Syk, p-BLNK|
|B cell||CD19, CD20, IgM, IgD, CD27, CD24||Oct-2, PAX-5, Blimp-1 ||IL-12, IFNy, IL-6, IL-10, TNFα||CD69, CD25, CD80, CD86|
|NK cell||CD45, CD56, CD16, absence of CD3||E4BP4, Eomes, T-bet, |
|IFNy, TNFα||CD161, NKp46, NKG2D, |
NKG2C, 2B4, NKp30, NKp44, CD69
|PD-1, TIM-3, TIGIT|
|Dendritic cells||CD11c, HLA-DR, CD123, CD1c/BDCA-1||BATF3, IRF4, IRF8||IFNβ, IL-12, IL-6, TNFα||CD80, CD86, CD40|
|Macro-phages||CD16, CD64, CD68, CD71, CCR5||iNOS, IRF4||IL-18, IL-10, TGFβ||CD54 (ICAM1), CD69, CD206, CD163, CD80, CD86|
|Tumor cells||EpCAM, E-cadherin, CD133, CD44, CD200, CD90, CD144, CD34, CD31||Lineage-specific cytokeratin subtypes|
In a regulated environment, analysis of such complex phenotypes needs to be standardized in a reproducible fashion. You can learn more about how Miltenyi Biotec has standardized flow cytometry-based immunophenotyping using highly characterized REAfinity™ Recombinant Antibodies, immunophenotyping kits, software compatibility, and flexible flow cytometers.
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