Development of a qPCR method to rapidly assess the function of NKT cells.
BACKGROUND
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.


OBJECTIVE
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.


CONCLUSIONS
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.
Department of Microbiology and Immunology, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Silke
S
Sohn
Department of Microbiology and Immunology, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Irina
I
Tiper
Department of Medicine, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Emily
E
Japp
Department of Microbiology and Immunology, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Wenji
W
Sun
Department of Medicine, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA.
Katherine
K
Tkaczuk
Department of Microbiology and Immunology, University of Maryland School of Medicine, Marlene and Stewart Greenebaum Cancer Center, Baltimore, MD 21201, USA. Electronic address: twebb@som.umaryland.edu.
Tonya J
TJ
Webb
D017951:Antigen Presentation; D055610:Antigens, CD1d / Q000378:metabolism; D001943:Breast Neoplasms / Q000276:immunology; D005260:Female; D006801:Humans; D016219:Immunotherapy, Adoptive; D007694:Killer Cells, Natural / Q000276:immunology*; D009369:Neoplasms / Q000276:immunology*; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*
D055610:Antigens, CD1d
CD1d; NKT cells; aAPCs