The TLR4 agonist, LPS and the TLR2/Dectin-1 agonist, Zymosan both potently induced G-CSF secretion by PBMCs, which was significantly suppressed by co-incubation with IFN-α (data not shown). As we found that PBMCs isolated from patients on IFN-α/ribavirin therapy did MK-8669 mw not secrete high levels of G-CSF (Fig. 1b), we wished to determine whether PBMCs isolated from these individuals could produce G-CSF in response to in vitro stimulation

with a TLR7/8 agonist that effectively drives G-CSF secretion by PBMCs (Fig. 3a). Therefore, we stimulated PBMCs isolated from HCV-infected patients receiving IFN-α/ribavirin therapy at week 24 of their treatment with CL097 and found that they secreted high levels of G-CSF in response to this stimulation (Fig. 4). Interferon-α has potent anti-viral activity and is the mainstay of anti-viral therapy for patients with chronic HCV infection. However, IFN-α has

significant toxic effects on the hematopoietic system. IFN-induced neutropenia frequently causes dose reduction or Adriamycin manufacturer treatment discontinuation. Bone marrow suppression contributes to the development of IFN-induced cytopenias.7 However, the effect of IFN-α on the expression of the hematopoietic growth factors that affect the development and efflux of neutrophils from bone marrow has not been studied in detail. G-CSF regulates neutrophil development. Mice lacking G-CSF have chronic neutropenia, granulocyte and macrophage progenitor cell deficiency and impaired neutrophil mobilization.17 Therefore, we investigated the effects of IFN-α on G-CSF production by PBMCs both in vitro and ex vivo from patients who had received therapeutic IFN-α to treat chronic HCV infection. We found that PBMCs isolated from patients on IFN-α/ribavirin therapy gradually lose the ability to produce G-CSF over the course of the treatment (Fig. 1b). The decline in the ability of patients’ PBMCs to produce G-CSF in culture paralleled the reduction in ANC over the course of IFN-α treatment, suggesting that suppressed G-CSF production by PBMCs may contribute to

click here IFN-α-induced neutropenia. Reduced G-CSF production by PBMCs may explain the suppressive effect of IFN-α on progenitor cell proliferation in bone marrow.7 The precise mechanism by which IFN-α exerts its suppressive effect on G-CSF production is unclear, in part because the mechanisms underlying the regulation of G-CSF production in vivo remain poorly defined.18 However, our finding that a TLR7/8 agonist induces G-CSF production in human monocytes suggests that NFκB has a role in the regulation of its expression. This is further confirmed by the recent finding that serum amyloid A (SAA) induces G-CSF production in mouse macrophages via a TLR2 dependent pathway.19 G-CSF stimulates angiogenesis and tumor growth.