The Synergistic Effect of Corticosteroids and Mycophenolic Acid on Chemokines in Orbital Cells From Patients With Graves’ Ophthalmopathy

In recent studies, an improvement of the response rate to therapy has been reported with corticosteroids and mycophenolic acid in patients with Graves’ ophthalmopathy (GO). In GO, retro-orbital cells (fibroblasts, preadipocytes, and extraocular muscle cells) secrete Th1 and Th2 chemokines stimulated by cytokines. Until now, no studies are present in literature regarding the effect of corticosteroids and mycophenolic acid on the secretion of chemokines in GO orbital cells. For this reason, the effect of increasing concentrations of mycophenolic acid or corticosteroids on the secretion of either the Th1 (CXCL10) and Th2 (CCL2) chemokines was tested in primary cultures of myoblasts, preadipocytes and fibroblasts obtained from GO patients. CXCL10 was undetectable in the supernatants of the retro-orbital cells in primary cultures; its release was induced dose-dependently by IFNγ, while TNFα alone had no effect. On the contrary CCL2 release (that was produced in low amounts basally) was dose-dependently induced by TNFα, while IFNγ alone had no effect. In both cases the combination of TNFα and IFNγ had a significant synergistic effect on CXCL10 and CCL2 secretion. The release of these chemokines was dose-dependently inhibited by increasing concentrations of mycophenolic acid, or corticosteroids (in a pharmacological range), in presence of IFNγ and TNFα stimulation. Moreover, the association of corticosteroids and mycophenolic acid (in presence of IFNγ and TNFα) had a stronger inhibitory effect on the chemokines release. In conclusion, in GO orbital cells, mycophenolic acid and/or corticosteroids (in a pharmacological range) have an inhibitory role on the secretion of both Th1 (CXCL10) and Th2 (CCL2) chemokines. This suggests a possible therapeutic role of these drugs.

Effect Of Granulocyte Colony-Stimulating Factor Mobilization On The Expression Of Th1/Th2 Chemokines and Their Receptors

Background Granulocyte colony-stimulating factor (G-CSF) mobilized peripheral blood stem cell (PBSC) has been used more frequently than bone marrow as the source of stem cells in allogeneic hematopoietic stem cell transplantation (allo-HSCT). Although it contains more mature T cells, neither the incidence nor the severity of acute graft-versus-host disease (aGVHD) is higher compared with bone marrow transplantation. This might be due to the immunoregulatory effects of G-CSF on adaptive immunity, including that G-CSF directly modulated via its receptor on T cells or indirectly modulated T cell immune responses via effector cells and cytokines. However, these mechanisms are not fully understood, and whether G-CSF could influence the expression of Th1/Th2 chemokines and their receptors remains unknown. The aim of this study is to investigate the effect of G-CSF mobilization on the expression of Th1/Th2 chemokines and their receptors. Methods The expression levels of Th1 chemokines (CXCL9, CXCL10, and CXCL11), Th2 chemokines (CCL17, CCL22) and their receptors CXCR3 and CCR4 were analyzed in peripheral blood mononuclear cells (PBMCs) from 25 donors before and after G-CSF mobilization, using real-time RT-PCR with SYBR Green±staining. The β2-microglobulin gene (β2-MG) was used as an endogenous reference, and the relative mRNA expression level of each gene was evaluated by the 2-ΔC t×100% method. Results The median expression level of CXCR3 was similar before and after G-CSF mobilization (0.1426% and 0.1109%) (P=0.278), while the expression level of CCR4 after G-CSF mobilization (0.0985%) was significantly lower than that before mobilization (0.1415%) (P=0.039). The median expression levels of CXCL9, CXCL10, and CXCL11 genes before mobilization (0.0048%, 0.0576% and 0.0079%) were not significantly different from that after G-CSF mobilization (0.0143%, 0.0666% and 0.0088%)(P=0.086, P=0.535 and P=0.680). The median expression levels of CCL17 and CCL22 were also similar before and after G-CSF mobilization (P=0.155, P=0.476). The expression pattern of three Th1 chemokines before mobilization was CXCL10> CXCL11> CXCL9, whereas it changed to CXCL10> CXCL9> CXCL11 after mobilization. The expression pattern of two Th2 chemokines before mobilization was CCL17> CCL22, whereas it changed to CCL22> CCL17 after mobilization. The relative expression levels of CXCL10 and CXCL11 genes before mobilization both showed a positive correlation to that after mobilization (P<0.001, r=0.760; P=0.024, r=0.470). Before mobilization, significant positive correlation was observed between the expression levels of CXCL9 and CXCL10, CXCL11 and CXCR3, CXCL11 and CCL22, CXCR3 and CCL22 (P<0.001, r=0.902; P=0.003, r=0.584; P=0.022, r=0.473; P<0.001, r=0.674, respectively). After G-CSF mobilization, significant positive correlation was observed between the expression levels of CXCL9 and CCL22, CXCL10 and CXCL11, CXCR3 and CCR4, CXCR3 and CCL22, CXCR3 and CCL17, CCL22 and CCL17 (P=0.001, r=0.653; P=0.001, r=0.665; P=0.002, r=0.602; P=0.028, r=0.458; P<0.001, r=0.738; P=0.044, r=0.424, respectively). Conclusions Our findings suggest that G-CSF mobilization might mainly influence the expression level of CCR4 genes in Th1/Th2 chemokines and their receptors. The expression patterns of Th1 chemokines and Th2 chemokines might both change after G-CSF mobilization. Disclosures: Li: This work was supported by Grants from National Natural Science Foundation of China (30871091 and 91129720), the Collaborated grant for HK-Macao-TW of Ministry of Science and Technology (2012DFH30060), the Guangdong Science & Technology Project (2012B0506: Research Funding. Liu:It was supported by 863 Program (No. 2011AA020105).: Research Funding; It was supported by National Public Health Grand Research Foundation (Grant No. 201202017), National Natural Science Foundation of China (Grant No.81000231, No.81270647).: Research Funding; It was supported by Science and Technology Program of Guangzhou of China (11A72121174).: Research Funding.

Bambusae caulisin Liquamen Suppresses the Expression of Thymus and Activation-Regulated Chemokine and Macrophage-Derived Chemokine in Human Keratinocytes due to Antioxidant Effect

Bambusae caulisin Liquamen (BCL), traditional herbal medicine used in East Asia, is known to have antioxidative and immune-regulating properties. We hypothesized that the potential antioxidant effects of BCL might suppress the production of thymus and activation-regulated chemokine (TARC) and macrophage-derived chemokine (MDC) in human keratinocytes (HaCaT cell). The immune-regulating effect of BCL was demonstrated by antioxidant capacity using DPPH analysis and DCFH-DA analysis. We found that BCL had strong ROS scavenge effect in HaCaT cell. BCL also showed suppression of IFN-γ-induced expression of TARC and MDC, activation of NF-κB, and, moreover, significant block of IFN-γ-induced degradation and phosphorylation of IκB. However, it had no effects on phosphorylation of p38 MAPK. Collectively, these results suggest that BCL may have a therapeutic potential on skin disease such as atopic dermatitis by inhibiting Th2 chemokines which is due, at least in part, to its antioxidant capacities.