Mesenchymal stem cells (MSCs) represent a population of cells which have the ability to regulate reactivity of T and B lymphocytes by multiple mechanisms. The immunoregulatory activities of MSCs are strictly influenced by the cytokine environment. Here we show that two functionally distinct cytokines, interleukin-4 (IL-4) and interferon-γ (IFN-γ), significantly potentiate the ability of MSCs to inhibit IL-10 production by activated regulatory B cells (Bregs). However, MSCs in the presence of IL-4 or IFN-γ inhibit the IL-10 production by different mechanisms. Preincubation of MSCs with IFN-γ led to the suppression, but pretreatment with IL-4 of neither MSCs nor B cells resulted in the suppression of IL-10 production. The search for candidate regulatory molecules expressed in cytokine-treated MSCs revealed different patterns of the gene expression. Pretreatment of MSCs with IFN-γ, but not with IL-4, induced expression of indoleamine-2,3-dioxygenase, cyclooxygenase-2 and programmed cell death-ligand 1. To identify the molecule(s) responsible for the suppression of IL-10 production, we used specific inhibitors of the putative regulatory molecules. We found that indomethacine, an inhibitor of cyclooxygenase-2 (Cox-2) activity, completely abrogated the inhibition of IL-10 production in cultures containing MSCs and IFN-γ, but had no effect on the suppression in cell cultures containing MSCs and IL-4. The results show that MSCs can inhibit the response of B cells to one stimulus by different mechanisms in dependence on the cytokine environment and thus support the idea of the complexity of immunoregulatory action of MSCs.
- Klíčová slova
- Cyclooxygenase 2, Cytokine environment, Gene expression, IFN-γ, IL-10, IL-4, Immunoregulation, Mesenchymal stem cells, Regulatory B cells,
- MeSH
- aktivace lymfocytů účinky léků imunologie MeSH
- antigeny CD279 genetika imunologie metabolismus MeSH
- buněčné mikroprostředí účinky léků imunologie MeSH
- cyklooxygenasa 2 genetika imunologie metabolismus MeSH
- cytokiny imunologie metabolismus farmakologie MeSH
- ELISA MeSH
- exprese genu účinky léků genetika imunologie MeSH
- indolamin-2,3,-dioxygenasa genetika imunologie metabolismus MeSH
- interferon gama farmakologie MeSH
- interleukin-10 imunologie metabolismus MeSH
- interleukin-4 farmakologie MeSH
- interleukin-6 genetika imunologie metabolismus MeSH
- kokultivační techniky MeSH
- kultivované buňky MeSH
- mezenchymální kmenové buňky účinky léků imunologie metabolismus MeSH
- myši MeSH
- polymerázová řetězová reakce s reverzní transkripcí MeSH
- regulační B-lymfocyty účinky léků imunologie metabolismus MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- antigeny CD279 MeSH
- cyklooxygenasa 2 MeSH
- cytokiny MeSH
- IL4 protein, human MeSH Prohlížeč
- indolamin-2,3,-dioxygenasa MeSH
- interferon gama MeSH
- interleukin-10 MeSH
- interleukin-4 MeSH
- interleukin-6 MeSH
- PDCD1 protein, human MeSH Prohlížeč
The immunoregulatory properties of mesenchymal stem cells (MSCs) have been well documented in various models in vitro and in vivo. Furthermore, a population of regulatory B cells (Bregs) that produce relatively high concentrations of IL-10 has been recently described. To study the relationship between MSCs and Bregs, we analyzed the effects of MSCs on IL-10 production by lipopolysaccharide (LPS)-activated mouse B cells. The production of IL-10 by B cells remained preserved in the presence of MSCs and was even significantly enhanced by IFN-γ. However, the production of IL-10 was strongly suppressed in cultures containing MSCs and IFN-γ. Preincubation of MSCs, but not of B cells, with IFN-γ induced the suppression of IL-10 secretion in cultures containing MSCs and B cells. The supernatants from IFN-γ-treated MSCs had no inhibitory effect, and the suppression of IL-10 production was abrogated if the MSCs and B cells were separated in a transwell system. Analysis of the gene expression of IFN-γ- or IFN-γ and LPS-treated MSCs revealed a strong upregulation of genes for indoleamine-2,3-dioxygenase (IDO), cyclooxygenase-2 (Cox-2) and programmed cell death-ligand 1 (PD-L1). While the inhibition of IDO activity or the inclusion of the neutralization monoclonal antibody anti-PD-L1 did not abrogate the suppression, indomethacin, an inhibitor of Cox-2, completely inhibited the MSC-mediated suppression of IL-10 production. Accordingly, the production of IL-10 by B cells was inhibited by exogenous prostaglandin E2. The results thus suggest that IFN-γ-treated MSCs strongly inhibit IL-10 production by activated B cells by a mechanism requiring cell contact and involving the Cox-2 pathway.
- Klíčová slova
- B cells, Cyclooxygenase-2, IL-10 production, Immunosuppression, Mesenchymal stem cells,
- MeSH
- aktivace lymfocytů účinky léků MeSH
- antigeny CD274 antagonisté a inhibitory genetika imunologie MeSH
- B-lymfocyty cytologie účinky léků imunologie MeSH
- cyklooxygenasa 2 genetika imunologie MeSH
- difuzní komory kultivační MeSH
- dinoproston farmakologie MeSH
- indolamin-2,3,-dioxygenasa genetika imunologie MeSH
- indomethacin farmakologie MeSH
- inhibitory cyklooxygenasy farmakologie MeSH
- interferon gama farmakologie MeSH
- interleukin-10 antagonisté a inhibitory genetika imunologie MeSH
- kokultivační techniky MeSH
- kultivační média speciální farmakologie MeSH
- lipopolysacharidy farmakologie MeSH
- mezenchymální kmenové buňky cytologie účinky léků imunologie MeSH
- mezibuněčná komunikace imunologie MeSH
- myši inbrední BALB C MeSH
- myši MeSH
- neutralizující protilátky farmakologie MeSH
- primární buněčná kultura MeSH
- regulace genové exprese MeSH
- signální transdukce MeSH
- zvířata MeSH
- Check Tag
- mužské pohlaví MeSH
- myši MeSH
- ženské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- antigeny CD274 MeSH
- Cd274 protein, mouse MeSH Prohlížeč
- cyklooxygenasa 2 MeSH
- dinoproston MeSH
- IL10 protein, mouse MeSH Prohlížeč
- indolamin-2,3,-dioxygenasa MeSH
- indomethacin MeSH
- inhibitory cyklooxygenasy MeSH
- interferon gama MeSH
- interleukin-10 MeSH
- kultivační média speciální MeSH
- lipopolysacharidy MeSH
- neutralizující protilátky MeSH
- Ptgs2 protein, mouse MeSH Prohlížeč
