Yanxia Zhan

The Ratio of Treg/Th17 Cells Correlates with the Disease Activity of Primary Immune Thrombocytopenia

Background Primary immune thrombocytopenia (ITP) is an autoimmune heterogeneous disorder that is characterized by decreased platelet count. Regulatory T (Treg) cells and T helper type 17 (Th17) cells are two subtypes of CD4+ T helper (Th) cells. They play opposite roles in immune tolerance and autoimmune diseases, while they share a common differentiation pathway. The imbalance of Treg/Th17 has been demonstrated in several autoimmune diseases. In this study, we aimed to investigate the ratio of the number of Treg cells to the number of Th17 cells in ITP patients and evaluate the clinical implications of the alterations in this ratio. Methods Thirty adult patients with newly diagnosed ITP enrolled in this study. Twelve patients had been clinically followed up for 12 months. The percentages of CD4+CD25hiFoxp3+ Treg cells and CD3+CD4+IL-17-producing Th17 cells in these patients and healthy controls (n = 17) were longitudinally analyzed by flow cytometry. Results The percentage of Treg cells in ITP patients was significantly lower than that of healthy controls, and the percentage of Th17 cells increased significantly at disease onset. The ratio of Treg/Th17 correlated with the disease activity. Conclusion The ratio of Treg/Th17 might be relevant to the clinical diversity of ITP patients, and this Treg/Th17 ratio might have prognostic role in ITP patients.

High-dose dexamethasone modulates serum cytokine profile in patients with primary immune thrombocytopenia.

Primary immune thrombocytopenia (ITP) is an autoimmune heterogeneous disorder which is characterized by decreased platelet count. Serum cytokines play an important role in the pathogenesis of ITP by initiating and perpetuating various cellular and humoral autoimmune processes. To investigate a broad spectrum of cytokines in ITP patients and the effects of high-dose dexamethasone (HD-DXM) regimen on serum cytokines profile, a multiplex cytokine assay was used to measure the serum levels of 20 circulating cytokines simultaneously in 22 patients before and after oral administration of 40mg/day DXM for 4 consecutive days. A cohort of 10 healthy individuals was served as control. Serum levels of interleukin (IL)-4, IL-5, IL-6, IL-10, IL-12p70, IL-13, growth-related oncogene (GRO), interferen (IFN)-γ and tumor necrosis factor (TNF)-α were significantly decreased in pre-treatment patients, compared with healthy controls (p<0.05). After HD-DXM treatment, IL-4, IL-5, IL-6, IL-12p70, IL-13, GRO, IFN-γ and TNF-α were significantly increased in remission patients as compared with patients before treatment (p<0.05). However, there was no significant difference (except TNF-α) between remission patients and healthy controls (p>0.05). All these cytokines decreased again in relapse patients. Our findings suggest that measuring cytokine levels might help in the clinical assessment of ITP, and the HD-DXM therapy could correct the derangement of serum cytokines.

Aberrant Frequency of IL-10-Producing B Cells and Its Association with Treg/Th17 in Adult Primary Immune Thrombocytopenia Patients

Background. Regulatory B cells (Breg) are a distinct B cell subset with immunoregulatory properties. Pivotal to Breg function is interleukin-10. This study was to investigate the role of IL-10-producing B cell (B10) and its association with Treg and Th17 subsets in immune thrombocytopenia (ITP) patients. Methods. Peripheral blood mononuclear cells from ITP patients and controls were stimulated with PMA, ionomycin, and Brefeldin A. The frequencies of CD19+IL-10+ B cells, CD3+CD4+IL-17+ Th17 cells, and CD4+CD25hiFoxp3+ Treg cells were analyzed by flow cytometry. The mRNA expression of Foxp3 and RORγt was detected by real-time quantitative PCR. Results. The number of B10 cells was elevated in ITP patients. After first-line therapies, it remained at high level in patients who achieved complete or partial response but decreased in those who acquired no response. There was a positive correlation between B10 cells and Tregs in ITP both before and after therapies. The ratio of Treg/Th17 decreased in ITP, and it strongly correlated with B10 cells. Conclusions. The frequency of B10 cells is elevated in ITP and it correlates with both the Tregs counts and the Treg/Th17 ratio. B10 cells to regulate functional T cell subsets might be impaired in patients with ITP.

Abnormal platelet kinetics are detected before the occurrence of thrombocytopaenia in HBV-related liver disease

Thrombocytopaenia is a frequent feature in patients with HBV‐related liver disease. Its underlying mechanism is not fully understood. Multiple factors might contribute to the development of thrombocytopaenia. In this study, we investigated the reticulated platelets (RP), glycocalicin (GC), serum thrombopoietin (TPO) and platelet glycoprotein (GP) in different stages of the disease.

The expression profile of toll-like receptor signaling molecules in CD19+ B cells from patients with primary immune thrombocytopenia

BACKGROUND B cells play a critical role in the pathogenesis of immune thrombocytopenia (ITP), and toll-like receptor (TLR) signaling is essential for the activation of autoreactive B cells. The objective of this study was to investigate the expression profile of TLR signaling molecules in circulating and splenic CD19(+) B cells isolated from ITP patients. METHODS CD19(+) B cells were magnetically isolated from peripheral blood and splenocytes. Human Toll-Like Receptor Signaling Pathway RT(2) Profiler™ PCR Array was used to determine the differences in mRNA expression of 84 TLR signaling pathway genes between ITP patients and controls. Flow cytometry was used to investigate intracellular expression of cytokines (IL-1β and IL-10). RESULTS A total of 31 genes involving TLR signaling pathways were differentially transcribed in circulating CD19(+) B cells, among which 27 were up-regulated in ITP. By comparison, differentially transcribed genes amounted to 39 in splenic B cells in ITP, among which only two were down-regulated. Up to 18 TLR signaling molecules exhibited up-regulated transcriptional levels both in splenic B cells and in circulating B cells in ITP. However, Only IL-10 and IL-1β were significantly upregulated in both the circulating and splenic B cells of patients with ITP. Intracellular staining of IL-10 and IL-1β confirmed the results of PCR Array. CONCLUSIONS The expression of TLRs and downstream cytokines, including IL-10 and IL-1β, is up-regulated in circulating and splenic B cells in ITP patients, suggesting that activated TLR signaling pathways in B cells may play dual roles in the pathophysiology of primary ITP.

Prolyl 4-hydroxylase 2 promotes B-cell lymphoma progression via hydroxylation of Carabin

B-cell lymphomas are heterogeneous blood disorders with limited therapeutic options, largely because of their propensity to relapse and become refractory to treatments. Carabin, a key suppressor of B-cell receptor signaling and proliferation, is inactivated in B-cell lymphoma by unknown mechanisms. Here, we identify prolyl 4-hydroxylase 2 (P4HA2) as a specific proline hydroxylase of Carabin. Carabin hydroxylation leads to its proteasomal degradation, thereby activating the Ras/extracellular signal-regulated kinase pathway and increasing B-cell lymphoma proliferation. P4HA2 is undetectable in normal B cells but upregulated in the diffuse large B-cell lymphoma (DLBCL), driving Carabin inactivation and lymphoma proliferation. Our results indicate that P4HA2 is a potential prognosis marker for DLBCL and a promising pharmacological target for developing treatment of molecularly stratified B-cell lymphomas.

Circulating cytokine portraits can differentiate between allograft rejection and pulmonary infection in cardiac transplant rats

OBJECTIVES Cardiac rejection and infection are the leading causes of morbidity and mortality after transplant representing with similar non-specific symptoms. Early discrimination is crucial yet challenging. We proposed that aberrant serum cytokine portraits exist in pulmonary infection and allograft rejection, and such profiles might aid in timely differential diagnosis. METHODS Lewis rat received Wistar rat heart allografts. Allograft rejection (n = 5) and pulmonary infection (n = 7) were induced via cessation of cyclosporine injection and intratracheal inoculation of Pseudomonas aeruginosa, respectively, and pathologically confirmed. A non-rejection and non-infection group (n = 5) was served as healthy controls. The circulating cytokine profiles of the study objects were then simultaneously measured using a multiplex quantitative cytokine array. RESULTS Thirteen cytokines [B7-2, β-nerve growth factor (NGF), chemokine (C-X3-C motif) ligand 1 (Fractalkine), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-γ), interleukin beta (IL-β), IL-2, IL-4, IL-10, chemokine (C-X-C motif) ligand 5 (LIX), L-selectin, chemokine (C-C motif) ligand 2 (MCP-1), brain creatine kinase (TCK-1) and tumour necrosis factor alpha (TNF-α)] were up-regulated in allograft rejecting animals. Among them, B7-2, β-NGF, Fractalkine, GM-CSF, IFN-γ, IL-β, IL-2, IL-4, LIX, MCP-1 and TCK-1 were significantly increased compared with infection rats (all P-values <0.05). B7-2, CNIC-1 and CNIC-2 were increased in infection animals when compared with healthy controls (900.85 ± 259.30 vs 175.04 ± 161.07 pg/ml, 319.68 ± 264.91 vs 13.50 ± 0.00 pg/ml and 51.424 ± 29.51 vs 5.24 ± 1.30 pg/ml, respectively, all P-values <0.05). CONCLUSIONS The present study demonstrated fluctuations in circulating cytokine portraits in cardiac allograft rejection and bacterial pulmonary infection after transplant. Such disease-specific cytokine patterns might facilitate early discrimination between rejection and infection.

Changes of Regulatory T and B Cells in Patients with Papillary Thyroid Carcinoma after 131I Radioablation: A Preliminary Study

Introduction. Lymphocytic infiltration and specific lymphocytes subsets may play important roles in papillary thyroid carcinoma (PTC) progression and prognosis. In this study, we try to understand the influence of 131I radioablation on the important lymphocytes subtypes of regulatory T and B cells (Tregs and Bregs). Methods. Peripheral blood mononuclear cells from 30 PTC patients before and after 131I therapy, and 20 healthy donors were collected. The expression of Tregs (CD4+CD25+CD127−/low) and B cell (CD5+CD19+) and production and secretion of interleukin 10 (IL-10) were analyzed by FACS and ELISA assay, respectively. Results. For Tregs percentage in peripheral blood lymphocytes, there was no difference between pretreatment and control and between posttreatment and control. Compared with pretherapy, increased Tregs infiltration was noted in posttherapy (P < 0.05). Although no difference was between pretreatment and control, compared with these two groups, decreased CD19+ and CD5+CD19+ B cell percentage in posttreatment was observed (P < 0.05). Among these groups, no significant difference was displayed in intracellular IL-10 production and extracellular IL-10 secretion. Conclusions. 131I Radioablation increased Tregs and decreased CD19+ and CD5+CD19+ B cells percentage after treatment. However, it has no effect on IL-10 and lymphocytes in peripheral blood. Therefore, longer follow-up of Tregs and Bregs should be further investigated.