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An Optimized Multicolor Immunophenotyping Panel to Profile NK Cells from Tumor Tissue
The History of NK Cells in Immuno-Oncology
Natural killer (NK) cells have been a focus in immuno-oncology for decades, largely due to their ability to destroy tumor cells through spontaneous cytotoxicity as well as their production of cytokines and chemokines that can help recruit additional immune cells.
Recent studies have shown that there is a correlation between the extent of NK cell infiltration and tumor progression.1,2
Learn more about NK cells and their role in immuno-oncology.
View NK cells in our interactive cell map
OMIP Panel Design
NK cells are notoriously difficult to study due to the limited amount of tissue that is normally available from tumor resections or biopsies.
Frutoso et al. developed an optimized multicolor immunophenotyping (OMIP) panel, which takes a flow cytometry immunophenotyping approach to extract as much information as possible from a single-cell suspension with the goal of providing in-depth characterization of human NK cells.3 Though other OMIP panels investigate these immune cells in a broader way (namely from human peripheral blood), this is the first specifically designed to profile NK cells isolated from tumor tissues.
In this paper, 3 the authors combined collagenase type II and DNase for tissue digestion to prepare the high-viability single-cell suspension, which yielded a good population of viable mucosal mononuclear cells when used with human mucosal tissues.
However, they found that detection of CD56, a biomarker typically used to identify NK cells, was affected substantially by the collagenase‐based tissue digestion. Thus, the panel is centered around the expression of NKp46, characterized as a highly conserved natural cytotoxicity receptor (NCR).
For flow cytometry immunophenotyping, the team used a 27-color phenotypic OMIP panel to identify NK cell activity and types. They compiled 18 biomarkers to assess NK cell function, including maturation, differentiation, migration, homing potential and functional state.
Watch our webinars on flow cytometry panel design
Performance of the OMIP Panel in Phenotyping NK Cells
Outside of the intended goal, most biomarker expression in this OMIP panel could be used to assess CD3+ T cells to help provide greater biological insights.
The team concluded that this specific combination of biomarkers and the flow cytometry immunophenotyping approach provided a thorough characterization of NK cells within tumor tissues and could prove to be a valuable tool in the study of immune cell function in human tumors and mucosal tissues.
Read the paper to learn more about this 27-color OMIP panel.
References
1.. Ishigami S, Natsugoe S, Tokuda K, Nakajo A, Che X, et al. Prognostic value of intratumoral natural killer cells in gastric carcinoma. Cancer 2000;88(3):577-583. doi: 10.1002/(SICI)1097-0142(20000201)88:3<577::AID-CNCR13>3.0.CO;2-V
2. Coca S, Perez-Piqueras J, Martinez D, Colmenarejo A, Saez MA, et al. The prognostic significance of intratumoral natural killer cells in patients with colorectal carcinoma. 2000;79(12):2320-2328. doi: 10.1002/(SICI)1097-0142(19970615)79:12<2320::AID-CNCR5>3.0.CO;2-P
3. Frutoso M, Mair F, Prlic M. OMIP‐070: NKp46‐Based 27‐Color Phenotyping to Define Natural Killer Cells Isolated from Human Tumor Tissues. Cytometry A. 2020;97(10): 1052-1056. doi: 10.1002/cyto.a.24230.