Maintenance of the blood system is dependent on dormant haematopoietic stem cells (HSCs) with long-term self-renewal capacity. After injury these cells are induced to proliferate to quickly re-establish homeostasis. The signalling molecules promoting the exit of HSCs out of the dormant stage remain largely unknown. Here we show that in response to treatment of mice with interferon-alpha (IFNalpha), HSCs efficiently exit G(0) and enter an active cell cycle. HSCs respond to IFNalpha treatment by the increased phosphorylation of STAT1 and PKB/Akt (also known as AKT1), the expression of IFNalpha target genes, and the upregulation of stem cell antigen-1 (Sca-1, also known as LY6A). HSCs lacking the IFNalpha/beta receptor (IFNAR), STAT1 (ref. 3) or Sca-1 (ref. 4) are insensitive to IFNalpha stimulation, demonstrating that STAT1 and Sca-1 mediate IFNalpha-induced HSC proliferation. Although dormant HSCs are resistant to the anti-proliferative chemotherapeutic agent 5-fluoro-uracil, HSCs pre-treated (primed) with IFNalpha and thus induced to proliferate are efficiently eliminated by 5-fluoro-uracil exposure in vivo. Conversely, HSCs chronically activated by IFNalpha are functionally compromised and are rapidly out-competed by non-activatable Ifnar(-/-) cells in competitive repopulation assays. Whereas chronic activation of the IFNalpha pathway in HSCs impairs their function, acute IFNalpha treatment promotes the proliferation of dormant HSCs in vivo. These data may help to clarify the so far unexplained clinical effects of IFNalpha on leukaemic cells, and raise the possibility for new applications of type I interferons to target cancer stem cells.

IntroductionThe incubation period of many viral infections is characterized by a transient reduction of peripheral blood lymphocyte counts, a phenomenon called lymphopenia. In mice infected with virus or treated with TLR ligands, lymphopenia in blood and lymph was related to IFN-␣/␤. 1,2 In line with this, lymphopenia is reported as a side effect of IFN-␣/␤ therapy for multiple sclerosis and chronic hepatitis. 3,4 Similarly, treatment with IFN-␥, IL-2, TNF-␣, 5 and IL-12 6 and chemical compounds including FTY720 7 were reported to cause a reduction of leukocyte subsets in patients' blood.Interferons are classified into 2 distinct families, IFN-␣/␤ (or type I IFN) and IFN-␥ (or type II IFN). IFN-␣/␤ binds to the heterodimeric type I interferon receptor (IFNAR), composed of an ␣-chain, which is essential for signaling, and a ␤-chain. In addition to antiviral activity, [8][9][10] IFN-␣/␤ can have an impact on lymphocyte proliferation, 11 apoptosis, 12 and expression of cytokines and cytokine receptors, [13][14][15][16] causing immune modulation in vivo. [17][18][19] IFN-␣/␤ exerts direct and indirect effects on lymphocytes. A critical role of IFN-␣/␤ in CD8 ϩ T-cell cross-priming has been shown. 20 This could recently be identified as a direct effect on CD8 ϩ T cells, promoting clonal expansion and memory T-cell formation by increased cell survival. 21 Furthermore, IFN-␣/␤ can directly stimulate naive T cells, 22,23 promote plasma cell differentiation, and enhance B-cell responses. [24][25][26][27][28] Many indirect effects of IFN-␣/␤ on lymphocytes are presumably mediated by the activation of antigen-presenting cells, 26,29-31 which in turn secrete various cytokines, such as IL-15, that can act on lymphocytes. 16 Apart from activating B and T cells, IFN-␣/␤ can influence lymphocyte homeostasis by suppressing hematopoiesis 32 or enhancing the output of lymphocyte precursors. 33 Lymphocytes continuously home to secondary lymphoid organs (SLOs) and recirculate through lymph and blood. Homing relies on a cascade of events guided by selectins, integrins, chemokine receptors, and sphingosine 1-phosphate receptor 1 (S1P 1 ). [34][35][36] The expression of homing receptors on lymphocytes and endothelium is stringently controlled and correlates with the cellular differentiation state. [37][38][39][40][41] It is conceivable that IFN-␣/␤ affects circulation and homing of lymphocytes because it can induce chemokines and modulate adhesion molecules in human T-cell lines 42 and endothelial cells. 43 Stimulation with the TLR7 ligand R-848 causes pronounced lymphopenia and enhances rolling and sticking of lymphocytes, concomitant with increased expression of several adhesion molecules on endothelia. 44 Furthermore, treatment with FTY720 reduces S1P 1 expression and leads to lymphopenia and lymphocyte sequestration in lymphoid organs. 7,45,46 In essence, however, the mechanism of lymphopenia and the cellular targets of cytokines and other factors remain elusive.In the present study, we have analyzed the role of IFN-␣/␤ in lymphope...

Despite development of new antiviral drugs, viral infections are still a major health problem. The most potent antiviral defense mechanism is the innate production of type I interferon (IFN-I), which not only limits virus replication but also promotes antiviral T cell immunity through mechanisms, which remain insufficiently studied. Using the murine lymphocytic choriomeningitis virus model system, we show here that IFN-I signaling on T cells prevented their rapid elimination in vivo. Microarray analyses uncovered that IFN-I triggered the expression of selected inhibitory NK-cell-receptor ligands. Consequently, T cell immunity of IFN-I receptor (IFNAR)-deficient T cells could be restored by NK cell depletion or in NK-cell-deficient hosts (Nfil3(-/-)). The elimination of Ifnar1(-/-) T cells was dependent on NK-cell-mediated perforin expression. In summary, we identified IFN-I as a key player regulating the protection of T cells against regulatory NK cell function.

FHL2 is a LIM-domain protein expressed in myoblasts but down-regulated in malignant rhabdomyosarcoma cells, suggesting an important role of FHL2 in muscle development. To investigate the importance of FHL2 during myoblast differentiation, we performed a yeast two-hybrid screen using a cDNA library derived from myoblasts induced for differentiation. We identified β-catenin as a novel interaction partner of FHL2 and confirmed the specificity of association by direct in vitro binding tests and coimmunoprecipitation assays from cell lysates. Deletion analysis of both proteins revealed that the NH2-terminal part of β-catenin is sufficient for binding in yeast, but addition of the first armadillo repeat is necessary for binding FHL2 in mammalian cells, whereas the presence of all four LIM domains of FHL2 is needed for the interaction. Expression of FHL2 counteracts β-catenin–mediated activation of a TCF/LEF-dependent reporter gene in a dose-dependent and muscle cell–specific manner. After injection into Xenopus embryos, FHL2 inhibited the β-catenin–induced axis duplication. C2C12 mouse myoblasts stably expressing FHL2 show increased myogenic differentiation reflected by accelerated myotube formation and expression of muscle-specific proteins. These data imply that FHL2 is a muscle-specific repressor of LEF/TCF target genes and promotes myogenic differentiation by interacting with β-catenin.

Although MERS-CoV has not yet acquired extensive distribution, being mainly confined to the Arabic and Korean peninsulas, it could adapt to spread more readily among humans and thereby become pandemic. Therefore, the development of a vaccine is mandatory. The integration of antigen-coding genes into recombinant MV resulting in coexpression of MV and foreign antigens can efficiently be achieved. Thus, in combination with the excellent safety profile of the MV vaccine, recombinant MV seems to constitute an ideal vaccine platform. The present study shows that a recombinant MV expressing MERS-S is genetically stable and induces strong humoral and cellular immunity against MERS-CoV in vaccinated mice. Subsequent challenge experiments indicated protection of vaccinated animals, illustrating the potential of MV as a vaccine platform with the potential to target emerging infections, such as MERS-CoV.

Activation of the cannabinoid 2 receptor (CB(2)) reduces ischemic injury in several organs. However, the mechanisms underlying this protective action are unclear. In a mouse model of ischemic stroke, we show that the CB(2) agonist JWH-133 (1 mg . kg(-1) . d(-1)) decreases the infarct size measured 3 d after onset of ischemia. The neuroprotective effect of JWH-133 was lost in CB(2)-deficient mice, confirming the specificity of JWH-133. Analysis of bone marrow chimeric mice revealed that bone marrow-derived cells mediate the CB(2) effect on ischemic brain injury. CB(2) activation reduced the number of neutrophils in the ischemic brain as shown by FACS analysis and by measuring the levels of the neutrophil marker enzyme myeloperoxidase. Indeed, we found in vitro that CB(2) activation inhibits adherence of neutrophils to brain endothelial cells. JWH-133 (1 microM) also interfered with the migration of neutrophils induced by the endogenous chemokine CXCL2 (30 ng/ml) through activation of the MAP kinase p38. This effect on neutrophils is likely responsible for the neuroprotection mediated by JWH-133 because JWH-133 was no longer protective when neutrophils were depleted. In conclusion, our data demonstrate that by activating p38 in neutrophils, CB(2) agonists inhibit neutrophil recruitment to the brain and protect against ischemic brain injury.-Murikinati, S., Jüttler, E., Keinert, T., Ridder, D. A., Muhammad, S., Waibler, Z., Ledent, C., Zimmer, A., Kalinke, U., Schwaninger, M. Activation of cannabinoid 2 receptors protects against cerebral ischemia by inhibiting neutrophil recruitment.

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Production of type I interferons (IFN-I, mainly IFNα and IFNβ) is a hallmark of innate immune responses to all classes of pathogens. When viral infection spreads to lymphoid organs, the majority of systemic IFN-I is produced by a specialized “interferon-producing cell” (IPC) that has been shown to belong to the lineage of plasmacytoid dendritic cells (pDC). It is unclear whether production of systemic IFN-I is generally attributable to pDC irrespective of the nature of the infecting pathogen. We have addressed this question by studying infections of mice with the intracellular bacterium Listeria monocytogenes. Protective innate immunity against this pathogen is weakened by IFN-I activity. In mice infected with L. monocytogenes, systemic IFN-I was amplified via IFN-β, the IFN-I receptor (IFNAR), and transcription factor interferon regulatory factor 7 (IRF7), a molecular circuitry usually characteristic of non-pDC producers. Synthesis of serum IFN-I did not require TLR9. In contrast, in vitro–differentiated pDC infected with L. monocytogenes needed TLR9 to transcribe IFN-I mRNA. Consistent with the assumption that pDC are not the producers of systemic IFN-I, conditional ablation of the IFN-I receptor in mice showed that most systemic IFN-I is produced by myeloid cells. Furthermore, results obtained with FACS-purified splenic cell populations from infected mice confirmed the assumption that a cell type with surface antigens characteristic of macrophages and not of pDC is responsible for bulk IFN-I synthesis. The amount of IFN-I produced in the investigated mouse lines was inversely correlated to the resistance to lethal infection. Based on these data, we propose that the engagement of pDC, the mode of IFN-I mobilization, as well as the shaping of the antimicrobial innate immune response by IFN-I differ between intracellular pathogens.