Abstract
For many years, high-affinity subunit of IL-2 receptor (CD25) has been considered as a promising therapeutic target for different pathologic conditions like allograft rejection, autoimmunity, and cancers. Although CD25 is transiently expressed by newly-activated T cells, it is the hallmark of regulatory T (Treg) cells which are the most important immunosuppressive elements in tumor microenvironment. Thus, Tregs can be considered as a potential target for chimeric antigen receptor (CAR)-based therapeutic approaches. On the other hand, due to some profound adverse effects pertaining to the use of CAR T cells, CAR NK cells have caught researchers’ attention as a safer choice. Based on these, the aim of this study was to design and develop a CAR NK cell against CD25 as the most prominent biomarker of Tregs with the prospect of overcoming immune escape mechanism in solid and liquid cancers. In the current study, an anti-CD25 CAR was designed and evaluated by comprehensive in silico analyses. Then, using lentiviral transduction system, NK-92 cell line was engineered to express this anti-CD25 CAR construct. In vitro functional analyses of anti-CD25 CAR for its reactivity against CD25 antigen as well as for cytotoxicity and cytokine production assays against CD25 bearing Jurkat cell line were done. In silico analyses demonstrated that the anti-CD25 CAR transcript and scFv protein structures were stable and had proper interaction with the target. Also, in vitro analyses showed that the anti-CD25 CAR-engineered NK-92 cells were able to specifically detect and lyse target cells with an appropriate cytokine production and cytotoxic activity. To conclude, the results showed that this novel CAR NK cell is functional and warrant further investigations.
Highlights
The cytokine interleukin (IL)-2, called “T-cell growth factor,” is a potent stimulator for in vitro expansion of T cells
CD25 has been used as a promising therapeutic target for the treatment of wide-range of pathologic conditions such as organ transplantation, graft versus host disease (GVHD), autoimmunity, and cancers
The missing loops in CD25 structure were fixed by SWISS-MODEL web server
Summary
The cytokine interleukin (IL)-2, called “T-cell growth factor,” is a potent stimulator for in vitro expansion of T cells. Activation of Tcells via T-cell receptor (TCR) lead to expression of high levels of a high affinity IL-2 receptor (IL-2R) [1]. This receptor is composed of three noncovalently bound subunits including an a chain ( called CD25), a b and a a chain. The role of IL-2 in the development and homeostasis of Tregs is critical and most of these cells express high levels of CD25 on their surface [5]. Tregs inhibit the expansion of effector T cells in vivo and suppress the autoimmune reactions as well as tumor immune surveillance, and antitumor immune responses. CD25 has been used as a promising therapeutic target for the treatment of wide-range of pathologic conditions such as organ transplantation, graft versus host disease (GVHD), autoimmunity, and cancers
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