Extracellular purine metabolism and signaling of CD73-derived adenosine in murine Treg and Teff cells. Am J Physiol Cell Physiol.

Department of Cardiovascular Physiology, Heinrich-Heine-University Duesseldorf, Duesseldorf, Germany.
AJP Cell Physiology (Impact Factor: 3.78). 05/2011; 301(2):C530-9. DOI: 10.1152/ajpcell.00385.2010
Source: PubMed


CD73-derived adenosine acts as potent inhibitor of inflammation, and regulatory T cells (Treg) have been shown to express CD73 as a novel marker. This study explored the role of endogenously formed adenosine in modulating NF-κB activity and cytokine/chemokine release from murine Treg and effector T cells (Teff) including key enzymes/purinergic receptors of extracellular ATP catabolism. Stimulating murine splenocytes and CD4(+) T cells with anti-CD3/anti-CD28 significantly upregulated activated NF-κB in CD73(-/-) T cells (wild type: 4.36 ± 0.21; CD73(-/-): 6.58 ± 0.75; n = 4; P = 0.029). This was associated with an augmented release of proinflammatory cytokines IL-2, TNF-α, and IFN-γ. Similar changes were observed with the CD73 inhibitor APCP (50 μM) on NF-κB and IFN-γ in wild-type CD4(+) T-cells. Treatment of stimulated CD4(+) T-cells with adenosine (25 μM) potently reduced IFN-γ release which is mediated by adenosine A2a receptors (A2aR). AMP (50 μM) also reduced cytokine release which was not inhibited by APCP. In Teff, A2aR activation (CGS21680) potently inhibited the release of IL-1, IL-2, IL-3, IL-4, IL-12, IL-13, IFN-γ, TNF-α, granulocyte-macrophage colony-stimulating factor (GM-CSF), CCL3, and CCL4. However, in Treg, CGS21680 did not alter cytokine/chemokine release. In summary, CD73-derived adenosine tonically inhibits active NF-κB in CD4(+) T-cells, thereby modulating the release of a broad spectrum of proinflammatory cytokines and chemokines. Downregulation of P2X7 and upregulation of CD73 in Treg after antigenic stimulation may be an important mechanism to maintain the ability of Treg to generate immunosuppressive adenosine.

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    • "Deficiency of CD73 on immune cells and thus the lack of CD73-generated adenosine was associated with delayed resolution of inflammation and adverse remodeling [19]. We have also shown that CD73-derived adenosine tonically inhibits active NF-κB in regulatory T and effector T cells, thereby modulating the release of a broad spectrum of pro-inflammatory cytokines and chemokines [20]. Since T lymphocytes orchestrate inflammatory responses and contribute to the resolution of inflammation [16], [17] this suggests a crucial role of T cell subsets and in particular adenosine in the healing process of ALI. "
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    ABSTRACT: Extracellular nucleotides and nucleosides have been implicated as important signaling molecules in the pathogenesis of acute lung injury (ALI). While adenosine is known to inhibit T cell activation, little information is available as to ATP and NAD degrading enzymes, the expression of ATP and adenosine receptors/transporters in different T cell subsets. ALI was induced by challenging mice with intra-tracheal instillation of 60 µl (3 µg/g) LPS. After 3 d and 7 d blood, lung tissue and bronchoalveolar lavage was collected and immune cells were analyzed using flow cytometry. The transcriptional phenotype of T helper cells, cytotoxic and regulatory T cells sorted by FACS was assessed by measuring the expression profile of 28 genes related to purinergic signaling using TaqMan Array Micro Fluidic Cards. Catabolism of ATP, NAD and cAMP by activated CD4+ T cells was evaluated by HPLC. CD73 was found to be highly abundant on lymphoid cells with little abundance on myeloid cells, while the opposite was true for CD39. After ALI, the abundance of CD39 and CD73 significantly increased on all T cell subsets derived from lung tissue and bronchoalveolar space. Expression analysis in T cell subsets of the lung revealed ATP (Cd39, Cd73) and NAD (Cd38, Cd157, Cd296, Pc-1) degrading enzymes. However, only transcription of Cd38, Cd39, Cd73, Ent1 and A2a receptor was significantly upregulated after ALI in T helper cells. CD4+ T cells from injured lung rapidly metabolized extracellular ATP to AMP and adenosine but not NAD or cAMP. These findings show that lung T cells - the dominant cell fraction in the later phase of ALI - exhibit a unique expression pattern of purinergic signaling molecules. Adenosine is formed by T cells at an enhanced rate from ATP but not from NAD and together with upregulated A2a receptor is likely to modulate the healing process after acute lung injury.
    PLoS ONE 04/2014; 9(4):e95382. DOI:10.1371/journal.pone.0095382 · 3.23 Impact Factor
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    • "CD73 and CD39 are expressed by CD4+ CD25+ FoxP3+ T regulatory cells in mice [11], [12] and humans [13]. Extracellular adenosine generated by these T regulatory cells binds A2a receptors on activated effector T cells suppressing proliferation [11], [14]. CD73 and CD39 are overexpressed in many cancer cells and function to suppress anti-tumor T cell responses via their adenosine production [15]–[17]. "
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    ABSTRACT: CD73 catalyzes the conversion of extracellular nucleosides to adenosine, modulating inflammatory and T cell responses. Elevated expression of CD73 marks subpopulations of murine memory B cells (MBC), but its role in memory development or function is unknown. Here, we demonstrate that CD73 is progressively upregulated on germinal center (GC) B cells following immunization, is expressed at even higher levels among T follicular helper cells, but is absent among plasma cells (PC) and plasmablasts (PB). We analyzed the T-dependent B cell response in CD73 knockout mice (CD73KO). During the early response, CD73KO and wild type (WT) mice formed GCs, MBCs and splenic PBs and PCs similarly, and MBCs functioned similarly in the early secondary response. Late in the primary response, however, bone marrow (BM) PCs were markedly decreased in CD73KO animals. Tracking this phenotype, we found that CD73 expression was required on BM-derived cells for optimal BM PC responses. However, deletion of CD73 from either B or T lymphocytes alone did not recapitulate the phenotype. This suggests that CD73 expression is sufficient on either cell type, consistent with its function as an ectoenzyme. Together, these findings suggest that CD73-dependent adenosine signaling is prominent in the mature GC and required for establishment of the long-lived PC compartment, thus identifying a novel role for CD73 in humoral immunity.
    PLoS ONE 03/2014; 9(3):e92009. DOI:10.1371/journal.pone.0092009 · 3.23 Impact Factor
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    • "Adenosine derived from the enzymatic breakdown of ATP by CD39 and CD73 mediates a considerable portion of the anti-inflammatory activities of Treg cells (Deaglio et al., 2007). Romio et al. showed that adenosine produced by Treg cells in concert with A2A receptors downregulated nuclear factor- κB (NF- κB) activation in T effector cells, which in turn reduced the release of proinflammatory cytokines and chemokines (Romio et al., 2011). Activation of the A2A receptor on Treg cells promotes the expansion of these cells, thereby increasing immune regulation (Ohta et al., 2012). "
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    ABSTRACT: In the last decade several groups have determined the key role of hemichannels formed by pannexins or connexins, extracellular ATP and purinergic receptors in physiological and pathological conditions. Our work and the work of others, indicate that the opening of Pannexin-1 hemichannels and activation of purinergic receptors by extracellular ATP is essential for HIV infection, cellular migration, inflammation, atherosclerosis, stroke, and apoptosis. Thus, this review discusses the importance of purinergic receptors, Panx-1 hemichannels and extracellular ATP in the pathogenesis of several human diseases and their potential use to design novel therapeutic approaches.
    Frontiers in Physiology 03/2014; 5:96. DOI:10.3389/fphys.2014.00096 · 3.53 Impact Factor
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