Thrombopoietin receptor agonists shift the balance of Fcγ receptors toward inhibitory receptor IIb on monocytes in ITP.

Elevated expression of the activating Fcγ receptor (FcγR) I and FcγRIIa together with decreased expression of the inhibitory FcγRIIb are involved in the pathogenesis of primary immune thrombocytopenia (ITP). Thrombopoietin receptor agonists (TPO-RAs) have been used clinically for the management of ITP; however, little is known about the effect of TPO-RAs on FcγR modulation in ITP. In this prospective study, we measured the alteration in monocyte FcγR expression from 21 corticosteroid-resistant/relapsed patients with chronic ITP receiving eltrombopag therapy. Results showed that the mRNA and protein levels of FcγRIIb were significantly elevated after 6-week eltrombopag treatment. Concurrently, FcγRI and IIa levels decreased remarkably, whereas FcγRIII expression did not change. In vitro phagocytosis assays indicated that a shift in the balance of FcγR toward inhibitory FcγRIIb on monocytes was accompanied with a considerable decrease in monocyte/macrophage phagocytic capacity. The response to eltrombopag therapy in patients with ITP was associated with FcγR phenotype and functional changes of monocytes/macrophages. Moreover, the plasma transforming growth factor-ß1 (TGF-ß1) concentrations increased significantly in eltrombopag responders. Modulation of monocyte FcγR balance by TPO-RAs was also found in a murine model of ITP established by transferring splenocytes from immunized CD61 knockout mice into CD61(+) severe combined immunodeficient mice. Romiplostim administration in ITP mice significantly upregulated inhibitory FcγRII expression and downregulated activating FcγRI expression. These findings showed that recovery of platelet counts after TPO-RA treatment in ITP is associated with the restoration of FcγR balance toward the inhibitory FcγRIIb on monocytes, and suggested that thrombopoietic agents have a profound effect on immune modulation in ITP. This study is registered at ClinicalTrials.gov as #NCT01864512.

High-dose dexamethasone corrects impaired myeloid-derived suppressor cell function via Ets1 in immune thrombocytopenia.

Myeloid-derived suppressor cells (MDSCs) are heterogeneous immature cells and natural inhibitors of adaptive immunity. In this study, the MDSC population was evaluated in adult patients with primary immune thrombocytopenia (ITP), where cell-mediated immune mechanisms are involved in platelet destruction. Our data demonstrated that both the numbers and suppressive functions of MDSCs were impaired in the peripheral blood and spleens of patients with ITP compared with healthy control patients. High-dose dexamethasone (HD-DXM) treatment rescued MDSC numbers in patients with ITP. And DXM modulation promoted the suppressive function of MDSCs induced in vitro. Moreover, the expression of interleukin 10 and transforming growth factor ß was significantly upregulated in DXM-modulated MDSCs compared with the unmodulated cultures. DXM-modulated MDSCs inhibited autologous CD4(+)T-cell proliferation and significantly attenuated cytotoxic T lymphocyte-mediated platelet lysis, further indicating enhanced control over T-cell responses. Elevated expression of the transcription factor Ets1 was identified in DXM-modulated MDSCs. Transfection of Ets-1 small interfering RNA efficiently blocked regulatory effects of MDSCs, which almost offset the augmentation of MDSC function by DXM. Meanwhile, splenocytes from CD61 knockout mice immunized with CD61(+)platelets were transferred into severe combined immunodeficient (SCID) mouse recipients (C57/B6 background) to induce a murine model of severe ITP. We passively transferred the DXM-modulated MDSCs induced from bone marrow of wild-type C57/B6 mice into the SCID mouse recipients, which significantly increased platelet counts in vivo compared with those receiving splenocyte engraftment alone. These findings suggested that impaired MDSCs are involved in the pathogenesis of ITP, and that HD-DXM corrected MDSC functions via a mechanism underlying glucocorticoid action and Ets1.

Cytokine changes in response to TPO receptor agonist treatment in primary immune thrombocytopenia.

Thrombopoietin receptor agonists (TPO-RAs) have been clinically used in primary immune thrombocytopenia (ITP) with favorable outcomes, while their effect on cytokine regulation in ITP remains unknown. In the present study, plasma and mRNA expression levels of interleukin (IL)-2, interferon gamma (IFN-γ), IL-4, IL-17A, and transforming growth factor-ß1 (TGF-ß1) were determined by ELISA and real-time quantitative PCR in 26 corticosteroid-resistant/relapsed ITP patients receiving eltrombopag or rhTPO therapy and 15 healthy controls (HCs). Results showed that plasma and mRNA levels of IL-2, IFN-γ, IL-4, and IL-17A in ITP patients did not change significantly after TPO-RA treatment, whereas TGF-ß1 levels increased remarkably. The pre- and post-treatment plasma and mRNA levels of IFN-γ and IL-2 were significantly higher, while the pre- and post-treatment IL-4 levels as well as the pre-treatment TGF-ß1 levels were remarkably lower in ITP patients compared with HCs. There was no significant difference in TGF-ß1 levels between TPO-RA-treated ITP patients and HCs. No statistical difference was found in plasma levels of IL-17A between ITP patients before or after treatment and HCs. However, the pre- and post-treatment mRNA expression of IL-17A and retinoic orphan receptor (ROR) γt in ITP patients were higher than that in HCs. Overall, these findings indicated that TPO-RA treatment could promote the secretion of TGF-ß1, while it could not correct the Th1 and Th17 polarization in ITP patients. This study might improve our understanding of the mechanism of action of TPO-RAs and provide important information for optimizing therapeutic strategies for ITP.

Interleukin 27 inhibits cytotoxic T-lymphocyte-mediated platelet destruction in primary immune thrombocytopenia.

Cytotoxic T-lymphocyte (CTL)-mediated platelet destruction and aberrant cytokine profiles play important roles in the pathogenesis of primary immune thrombocytopenia (ITP). Interleukin-27 (IL-27) has pleiotropic immunomodulatory effects. However, the effect of IL-27 on CTL activity in ITP has not been reported. In the present study, platelets from ITP patients were cultured with autologous CTLs in the presence of IL-27. We found that IL-27 could inhibit CTL-mediated platelet destruction. In these IL-27-treated CTLs, granzyme B and T-bet expression decreased significantly, whereas granzyme A, perforin, and eomesodermin were not affected. To further investigate the role of granzyme B in CTL-mediated platelet destruction, granzyme B inhibitor was added and platelet apoptosis was significantly inhibited. These results suggest that IL-27 negatively regulates CTL cytotoxicity toward platelets in ITP by decreasing granzyme B expression, which is associated with reduced T-bet expression. IL-27 may have a therapeutic role in treating ITP patients.

Distributed lag effects and vulnerable groups of floods on bacillary dysentery in Huaihua, China.

Understanding the potential links between floods and bacillary dysentery in China is important to develop appropriate intervention programs after floods. This study aimed to explore the distributed lag effects of floods on bacillary dysentery and to identify the vulnerable groups in Huaihua, China. Weekly number of bacillary dysentery cases from 2005-2011 were obtained during flood season. Flood data and meteorological data over the same period were obtained from the China Meteorological Data Sharing Service System. To examine the distributed lag effects, a generalized linear mixed model combined with a distributed lag non-linear model were developed to assess the relationship between floods and bacillary dysentery. A total of 3,709 cases of bacillary dysentery were notified over the study period. The effects of floods on bacillary dysentery continued for approximately 3 weeks with a cumulative risk ratio equal to 1.52 (95% CI: 1.08-2.12). The risks of bacillary dysentery were higher in females, farmers and people aged 15-64 years old. This study suggests floods have increased the risk of bacillary dysentery with 3 weeks' effects, especially for the vulnerable groups identified. Public health programs should be taken to prevent and control a potential risk of bacillary dysentery after floods.

[Responses of non-structural carbohydrate metabolism of cucumber seedlings to drought stress and doubled CO2 concentration].

The effects of doubled CO2 concentration on non-structural carbohydrate metabolism of cucumber (Cucumis sativus L. cv. 'Jinyou No.1') seedlings under drought stress were investigated. Split plot design was deployed, with two levels of CO2 concentrations (ambient CO2 concentration, 380 µmol . mol-1, and doubled CO2 concentration, 760±20 µmol . mol-1) in the main plots, and three levels of water treatments (control, moderate drought stress, and severe drought stress) simulated by PEG 6000 in the split-plots. The results showed that non-structural carbohydrates of cucumber leaves, including glucose, fructose, sucrose, and stachyose, increased with the doubling of CO2 concentration, which resulted in the decreased osmotic potential, improving the drought stress in cucumber seedlings. During the drought stress, sucrose synthase, soluble acid invertase and al- kaline invertase started with an increase, and followed with a decline in the leaves. In the root system, however, soluble acid invertase and alkaline invertase increased gradually throughout the whole process, whereas sucrose phosphate synthase firstly increased and then decreased. The treatment of doubled CO2 enhanced the activity of sucrose synthase, but decreased the activity of sucrose phosphate synthase. The synergistic action of the two enzymes and invertase accelerated the decomposition of sucrose and inhibited the synthesis of sucrose, leading to the accumulation of hexose, which lowered the cellular osmotic potential and enhanced the water uptake capacity. In conclusion, doubled CO2 concentration could alleviate the adverse effects of drought stress and improve the drought tolerance of cucumber seedlings. Such mitigating effect on cucumber was more significant under severe drought stress.