Abstract
Maintenance of cellular function in culture is vital for transfer and development following adoptive immunotherapy. Dual properties of IL-21 in activating T cells and reducing activation induced cell death led us to explore the mechanism of action of IL-21 enhanced proliferation and cytotoxic potential of CIK cells. CIK cells cultured from PBMCs of healthy subjects were stimulated with IL-21 and cellular viability and cytotoxicity to K562 cells were measured. To elucidate the mechanism of action of IL-21, mRNA expression of cytotoxic factors was assessed by RT-PCR and protein expression of significantly important cytotoxic factors and cytokine secretion were determined through flow cytometry and ELISA. Western blotting was performed to check the involvement of the JAK/STAT pathway following stimulation. We found that IL-21 did not enhance in vitro proliferation of CIK cells, but did increase the number of cells expressing the CD3+/ CD56+ phenotype. Cytotoxic potential was increased with corresponding increase in perforin (0.9831±0.1265 to 0.7592±0.1457), granzyme B (0.4084±0.1589 to 0.7319±0.1639) and FasL (0.4015±0.2842 to 0.7381±0.2568). Interferon gamma and TNF-alpha were noted to increase (25.8±6.1 ng/L to 56.0±2.3 ng/L; and 5.64±0.61 μg/L to 15.14±0.93 μg/L, respectively) while no significant differences were observed in the expression of granzyme A, TNF-alpha and NKG2D, and NKG2D. We further affirmed that IL-21 signals through the STAT-3 and STAT- 5b signaling pathway in the CIK cell pool. IL-21 enhances cytotoxic potential of CIK cells through increasing expression of perforin, granzyme B, IFN-gamma and TNF-alpha. The effect is brought about by the activation of STAT-3 and STAT-5b proteins.
Highlights
IL-21 was originally implicated as a regulator of T and B cell proliferation as well as of NK cell maturation
Was synthesized from 1ug of RNA in 4ul 5×RT Buffer IL-21 showed no considerable increase in the number of containing dNTP Mixture (10 mM) 2 μl, Random primer CIK cells we explored whether IL-21 could increase the
IL-21 like IL15 is unique among the common γchain family of cytokines in its ability to maintain CD8 T cell population without causing any activation induced cell death (Moroz et al, 2004)
Summary
IL-21 was originally implicated as a regulator of T and B cell proliferation as well as of NK cell maturation. It belongs to a subset of cytokines where the receptors share the common cytokine receptor γ chain, γc. The common cytokine receptor γc is mutated in X-linked severe combined immunodeficiency, a disease with severely impaired T cell and NK cell development and diminished B cell function. Γc is a critical component of the receptors for IL-2, IL-4, IL-7, IL-9, IL-15, and IL-21, which together regulate lymphocyte development and control a broad spectrum of activities that shape innate and acquired immune responses. Th17 cells, which are critical mediators of autoimmunity, depend on IL-21, together with IL-6 and TGF-β, for their differentiation
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
