Characterization of bovine interleukin-2 stably expressed in HEK-293 cells

Abstract

Interleukin 2 (IL-2) is a pleotropic cytokine and well-known as a T cell growth factor in immunology. It is now known to exert both immunostimulatory and immunosuppressive effects, optimizing immunological microenvironments for effector and regulatory T cell responses. The immunomodulatory role of IL-2 is critical for deciding whether or not T cell responses against specific antigens result in protection. We have established a mammalian cell line (HEK-293) stably expressing bovine IL-2 (boIL-2) (designated as HEK-293/boIL-2), using the piggyBac transposon system. The concentration of recombinant bovine IL-2 (rboIL-2) in the culture supernatant of HEK-293/boIL-2 reached 100 ng/ml on day 7 and showed similar proliferative activity to recombinant human IL-2 (rhuIL-2) for bovine peripheral mononuclear blood cells. Although rhuIL-2 has been often used to activate bovine T cells, our results indicate that characteristics of the T cell activation through rboIL-2 and huIL-2 appear slightly but significantly different. Interestingly, the rboIL-2/anti-boIL-2 monoclonal antibody (C5) (rboIL-2/C5) complex strongly induced proliferation of bovine NKp46 + cells, natural killer (NK) cells, in vitro . This indicates that the rboIL-2/C5 complex could function as an IL-2 agonist specifically to increase the NK cell population, which in turn could enhance the activity of NK cells leading to protective immunity.

Effects of mulberry leaf silage on antioxidant and immunomodulatory activity and rumen bacterial community of lambs

“…In addition, the IL-1β, a master proinflammatory cytokine, acts as an adjuvant to stimulate antigen-specific immune responses [ 47 ]. IL-2 and IL-6 has both pro- and anti-inflammatory activity [ 48 ]. Therefore, the much lower IL-1β and IL-6 in MS compared to AS group further confirmed the immunomodulating roles of mulberry leaf in Tan-lambs.…”

The detection of long‐lasting memory foot‐and‐mouth disease (FMD) virus serotype O‐specific CD4+ T cells from FMD‐vaccinated cattle by bovine major histocompatibility complex class II tetramer

“…PBMCs, plated at a density of 2·5 × 10 6 cells/ml in 48‐well plates, were stimulated by incubating with the BEI FMDV‐O1 M intact antigen at 0·1 µg/mL for six days at 37℃, 5% CO 2 . After six‐day culture, the PBMCs were transferred to new wells and treated with fresh cRPMI medium and 0·5 ng/ml of recombinant bovine interleukin‐2 (rboIL‐2, [ 29 ] every other day until flow cytometry analysis on days 12‐16. Prior to adding fresh medium and rboIL‐2, 500 µL of culture supernatant was gently removed from the surface without disturbing the cells.…”