Related Experiment Video
Updated: May 1, 2026
qKAT: Quantitative Semi-automated Typing of Killer-cell Immunoglobulin-like Receptor Genes
Published on: March 6, 2019
Development of a qPCR method to rapidly assess the function of NKT cells
Silke Sohn1, Irina Tiper1, Emily Japp2
1Department of Microbiology and Immunology, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Introduction:
NKT cells comprise a rare, but important subset of T cells which account for ~0.2% of the total circulating T cell population. NKT cells are known to have anti-tumor functions and rapidly produce high levels of cytokines following activation. Several clinical trials have sought to exploit the effector functions of NKT cells. While some studies have shown promise, NKT cells are approximately 50% lower in cancer patients compared to healthy donors of the same age and gender, thus limiting their therapeutic efficacy. These studies indicate that baseline levels of activation should be assessed before initiating an NKT cell based immunotherapeutic strategy.
Aim:
The goal of this study was to develop a sensitive method to rapidly assess NKT cell function.
Methods:
We utilized artificial antigen presenting cells in combination with qPCR in order to determine NKT cell function in peripheral blood mononuclear cells from healthy donors and breast cancer patients.
Results:
We found that NKT cell activation can be detected by qPCR, but not by ELISA, in healthy donors as well as in breast cancer patients following four hour stimulation.
Conclusion:
This method utilizing CD1d-expressing aAPCs will enhance our knowledge of NKT cell biology and could potentially be used as a novel tool in adoptive immunotherapeutic strategies.
Insights
A new qPCR method rapidly assesses Natural Killer T (NKT) cell function, even in cancer patients where NKT cell levels are lower. This aids in evaluating NKT cell-based immunotherapies.
Area of Science:
- Immunology
- Cell Biology
- Cancer Research
Background:
- Natural Killer T (NKT) cells are crucial for anti-tumor immunity but are reduced in cancer patients, limiting immunotherapy.
- Assessing baseline NKT cell activation is vital for effective NKT cell-based therapeutic strategies.
Purpose of the Study:
- To develop a sensitive and rapid method for assessing NKT cell function.
- To evaluate NKT cell activation in both healthy donors and breast cancer patients.
Main Methods:
- Utilized artificial antigen-presenting cells (aAPCs) engineered to express CD1d.
- Employed quantitative Polymerase Chain Reaction (qPCR) to detect NKT cell activation.
- Analyzed peripheral blood mononuclear cells (PBMCs) from healthy donors and breast cancer patients.
Main Results:
- NKT cell activation was successfully detected using qPCR after a four-hour stimulation.
- ELISA was unable to detect NKT cell activation under the same conditions.
- The qPCR method demonstrated sensitivity in detecting NKT cell function in both healthy and cancer patient samples.
Conclusions:
- The developed qPCR method using CD1d-expressing aAPCs is a sensitive tool for assessing NKT cell function.
- This method can enhance understanding of NKT cell biology.
- It holds potential as a novel diagnostic or monitoring tool for NKT cell-based immunotherapies.
