Production of IL-10 by Human Natural Killer Cells Stimulated with IL-2 and/or IL-12

Abstract

Human NK cell activity can be augmented in vitro by stimulation with IL-2 or IL-12, both of which also induce the production of IFN-γ, TNF-α, and granulocyte-macrophage CSF by NK cells. For the first time, we demonstrate that freshly purified NK cells stimulated with IL-2 proliferated and produced IL-10 in a dose-dependent manner. IL-10 mRNA expression, as detected by semiquantitative reverse transcription-PCR, reached peak levels at 24 h. IL-10 protein was detectable on day 2 and further increased on days 3 and 6 as measured by ELISA. However, IL-12 alone induced neither substantial proliferation nor detectable IL-10 production by fresh NK cells, but it synergized with IL-2 in inducing IL-10 mRNA expression and protein synthesis. IL-10 production by activated NK cells was confirmed by intracytoplasmic cytokine staining by three-color immunofluorescence of CD16+ and/or CD56+ NK cells with anti-IL-10 antibody. IL-10 production by NK cells was further confirmed in the NK-like cell line, YT, which constitutively expressed IL-10 mRNA and protein. IL-12 alone did not induce NK proliferation, but it inhibited IL-2-induced proliferation. Neutralization of endogenously produced IL-10 with anti-IL-10 antibodies did not overcome the inhibition of IL-2-induced proliferation by IL-12. Together, these results demonstrate that IL-2 and IL-12 synergize to induce IL-10 production by human NK cells and that IL-12 inhibits IL-2 induced NK cell proliferation by an IL-10-independent mechanism.

Circulating CXCR5+ natural killer cells are expanded in patients with myasthenia gravis

“…13 Moreover, NK cells are known to produce a variety of cytokines depending on the inflammatory microenvironment, such as IFN-γ, IL-4 and IL-10, and exert profound effects on the immune response. 3,14,15 Our previous results revealed a specific subpopulation of NK cells expressing CXCR5 and highlighted the different roles of CXCR5 + NK cells and CXCR5 -NK cells in EAMG. 6 However, whether and how these different NK cell subtypes are relevant to the pathogenesis of MG remains unclear.…”

“…In contrast, the minority population of circulating CD56 bright NK cells (10%) is generally considered cytokine‐producing regulatory cells 13 . Moreover, NK cells are known to produce a variety of cytokines depending on the inflammatory microenvironment, such as IFN‐γ, IL‐4 and IL‐10, and exert profound effects on the immune response 3,14,15 . Our previous results revealed a specific subpopulation of NK cells expressing CXCR5 and highlighted the different roles of CXCR5 + NK cells and CXCR5 − NK cells in EAMG 6 .…”

Disruption of Genes Encoding Putative Zwitterionic Capsular Polysaccharides of Diverse Intestinal Bacteroides Reduces the Induction of Host Anti-Inflammatory Factors

“…This might indicate that different ZPS have differential interactions with various cell populations. Indeed, PBMC represents a complex milieu of different cell types capable of producing IL-10 under certain conditions, including NK cells, monocytes, and dendritic cells [ 20 ]. In our previous work [ 11 ], we used a 24-h intracellular cytokine staining (ICCS) assay to evaluate IL-10 production by CD4 + T cells in stimulations of PBMC with wildtype B. fragilis versus B. fragilis ΔPSA.…”

Atividade imunomoduladora da alga Chlorella vulgaris em camundongos portadores de tumos ascitico de Ehrlich

“…A IL-10 foi inicialmente descrita como uma citocina imunoreguladora, que é produzida tardiamente após estimulação por células T e macrófagos (Yssel et al 1992). No entanto, distintas vias de sinalização que levam à produção de IL-10 por várias outras células, incluindo as células B, mastócitos e células NK, também foram mostrados (Howard et al 1992;Mehrotra et al 1998). Curiosamente, IL-10 foi mencionada por prejudicar a capacidade dos macrófagos de fornecer sinais de co-estimulação para a proliferação de células T, efeito este principalmente mediado pela inibição da produção de IL-2 (Parhar & Lala 1988).…”