Treg Cells In Peripheral Blood Research Articles

Clinical study reveals the efficacy of sirolimus in treating primary immune thrombocytopenia: findings from a single-center study.

Immune thrombocytopenia (ITP) is an autoimmune disease that arises because of self-destruction of circulating platelets. The mechanism remains complicated and lacks a standard clinical treatment. Current first-line and second-line medications for ITP have shown limited effectiveness, necessitating the exploration of new therapeutic options. Sirolimus is a mammalian target of rapamycin (mTOR) inhibitor that has been demonstrated to inhibit lymphocyte activity, indicating potential for SRL in the treatment of ITP. This study aimed to evaluate the clinical efficacy of sirolimus as a second-line drug in patients with ITP. The starting dose of sirolimus for adults ranged from 2 to 4 mg/day, with a maintenance dose of 1 to 2 mg/day. For children, the starting dose was 1-2 mg/day, with a maintenance dose of 0.5-1 mg/day. The dosage could be adjusted if needed to maintain a specific blood concentration of sirolimus, typically between 5 and 15 ng/ml, throughout the treatment period. After 3 months, the overall response rate was 60% (12/20), with 30% of patients (6/20) achieving a complete response (CR) and 30% (6/20) achieving a partial response (PR). The CR rate at 6 months remained consistent with the 3-month assessment. No major adverse events were reported, indicating that sirolimus was well tolerated and safe. Analysis of peripheral blood Treg cell percentages in both the control and ITP showed no significant difference before treatment. The percentage of Treg cells increased after treatment with sirolimus, suggesting that sirolimus increases Treg cells. These findings suggest that sirolimus serves as an effective second-line treatment option for ITP, demonstrating favorable clinical efficacy.

Rituximab may affect T lymphocyte subsets balance in primary membranous nephropathy

BackgroundThe aim of this study was to investigate the effects and significance of rituximab (RTX) on the levels of T lymphocyte subsets in patients diagnosed with primary membranous nephropathy (PMN).MethodsA total of 58 PMN patients and 25 healthy donors were chosen as the subjects. Among the PMN patients, 40 individuals received RTX treatment and completed at least 6 months of follow-up. All subjects underwent flow cytometry analysis to determine the peripheral blood lymphocyte subsets. The changes in anti-PLA2R antibody titers and 24-hour urinary protein levels were evaluated by ELISA and Biuret method before and after treatment.Results(1) The PMN group exhibited a significantly greater percentage of peripheral blood CD3−CD19+ B cells than the healthy group, which is consistent with the findings of previous reports. Additionally, compared with those in the peripheral blood of healthy individuals, the numbers of CD4+ central memory T cells, CD4+ effector memory T cells, CD4+/CD8+, and CD4+CD25+ T cells in the PMN peripheral blood were markedly greater. However, the number of peripheral blood Treg cells was reduced in the PMN group. (2) After 6 months of RTX treatment, PMN patients exhibited significant decreases in anti-PLA2R antibody titers, 24-hour urinary protein levels, and peripheral blood CD3−CD19+ B cells. Importantly, RTX administration decreased CD4+CD25+ T cells and CD4+/CD8+ in the peripheral blood of PMN patients and improved Treg cell levels. (3) RTX treatment induced alterations in the CD4+ T lymphocyte subsets in PMN patients, which did not correlate with B lymphocyte counts or anti-PLA2R antibody titers.ConclusionsRTX treatment might have a beneficial impact on cellular immunity by effectively restoring the balance of CD4+ T lymphocyte subsets in PMN patients, which is beyond its effects on B cells and antibody production.Trial registrationThe research was registered at the First Affiliated Hospital of Soochow University. Registration Number: MR-32-23-016211. Registration Date: May 31, 2023.

Sulforaphane effectively inhibits HBV by altering Treg/Th17 immune balance and the MIF-macrophages polarizing axis in vitro and in vivo

BackgroundHepatitis B virus (HBV) infection is a major public health problem. After HBV infection, viral antigens shift the immune balance in favor of viral escape. Sulforaphane (SFN) is a traditional Chinese medicine.It regulates multi-biological activities, including anti-inflammation, anticancer, and antiviral. However, few studies reported that SFN can inhibit HBV infection before. MethodsAn immunocompetent HBV CBA/CaJ mouse model and a co-culture model were used to explore the effect of SFN on HBV and whether SFN altered the immune balance after HBV infection. ResultsWe found that SFN was able to reduce HBV DNA, cccDNA, HBsAg, HBeAg, and HBcAg levels in serum and liver tissues of HBV-infected mice. In vitro and in vivo experiments showed that SFN could significantly increase the expression of Cd86 and iNOS and inhibit the expression of Arg1 on macrophages after HBV infection. After SFN administration, Th17 markers in liver tissue and serum were significantly increased. There was no significant changes in the proportion of Treg cells in peripheral blood, but a significant increase in the proportion of Th17 cells and decrease of the Treg/Th17 ratio. Using a network pharmacology approach, we predicted macrophage migration inhibitory factor (MIF) as a potential target of SFN and further validated that MIF expression was significantly increased after HBV infection and SFN significantly inhibited MIF expression both in vitro and in vivo. There was an upward trend in HBV markers (p>0.05) after MIF overexpression. Overexpression of MIF combined with the use of SFN resulted in a significant reversion in the expression of HBV markers and polarization of macrophages towards the M1 phenotype. ConclusionOur results indicated that immunocompetent HBV CBA/CaJ mouse model is a good model to evaluate HBV infection. SFN could inhibit the expression of HBV markers, promote polarization of macrophages towards the M1 phenotype after HBV infection, change the proportion of Treg and Th17 cells. Our findings demonstrate that SFN inhibit HBV infection by inhibiting the expression of MIF and promoting the polarization of macrophages towards the M1 phenotype, which illustrates a promising therapeutic approach in HBV infection.

MicroRNA-155 is a critical regulator of regulatory T cells in OM-85 Broncho-Vaxom treated experimental models of allergic rhinitis

<abstract><sec> <title>Background</title> <p>Bacterial lysates could alleviate the clinical symptoms of allergic rhinitis (AR) and decrease the recurrent rate of AR through regulation of regulatory T cell (Treg) cells. However, the molecular regulatory mechanisms of bacterial lysates to Treg are still unclear.</p> </sec><sec> <title>Objective</title> <p>We aimed to investigate the importance of microRNA-155 (miR-155) to Treg cells function in OM-85 Broncho-Vaxom (OM-85 BV) treated experimental mouse models of AR.</p> </sec><sec> <title>Methods</title> <p>AR mouse models were established and treated by intranasal administration of OM-85 BV to investigate the role of bacteria lysate for Treg cell function. The proliferation of Treg cells in peripheral blood was examined. The mRNA levels of IL-10, transforming growth factor-β (TGF-β) were examined by real-time PCR. miR-155 mimics and inhibitor were used to verify the role of miR-155 for Treg cells function.</p> </sec><sec> <title>Results</title> <p>OM-85 BV, miR-155 mimics or their combination reduced total cells, lymphocytes, neutrophils and eosinophils in nasal lavage fluid of AR mouse models and improved allergic symptoms. OM-85 BV promoted the proliferation of Treg and the expression of Foxp3, IL-10 and TGF-β both <italic>in vivo</italic> and <italic>in vitro</italic>. The miR-155 enhanced the proliferation and function of Treg.</p> </sec><sec> <title>Conclusions</title> <p>MiR-155 promotes Treg cells function in OM-85 BV bacteria lysate treated experimental models of AR and alleviate the upper airway allergic inflammation in AR mice.</p> </sec></abstract>

Reproductive Immunology and Embryo Acceptance

Pregnancy is a test of the fundamental laws of transplantation immunology - the coexistence of genetically disparate tissues without host immune rejection. Advances over recent decades show that implantation and placental development depend on an active state of maternal immune tolerance, and breakdown of tolerance is a key contributor to infertility and recurrent pregnancy loss. Tolerance in pregnancy depends on regulatory T (Treg) cells, which are critical to inhibit effector immunity, constrain inflammation, and support maternal vascular adaptations that allow placental access to maternal nutrient supply. Treg cell defects are implicated in disorders of embryo implantation and placental development, but the origins of Treg cell dysfunction are unknown. In recent studies, we have comprehensively analysed the phenotypes and transcriptional profile of peripheral blood Treg cells in individuals with early pregnancy failure (recurrent miscarriage and recurrent implantation failure). Compared to fertile women, women with early pregnancy failure have fewer total Treg cells and altered Treg cell phenotypes, accompanied by lower Treg:Th1 and Treg:Th17 ratios. RNAseq demonstrates an aberrant Treg cell gene expression profile in early pregnancy failure, with upregulation of pro-inflammatory genes including CSF2, IL4, IL17A, IL21, and IFNG. Our data indicate that Treg cell defects in women with early pregnancy failure can arise due to loss of lineage fidelity associated with impaired FOXP3 regulation. Factors including metabolic function, luteal phase progesterone, and male partner seminal fluid components are all implicated in causing Treg cell deficit. Treg cells may be a useful target for therapeutic interventions to improve uterine receptivity, but this will depend on developing diagnostic tests that are informative. Moreover, safe, and effective interventions to modulate these cells are in their infancy, and personalized approaches matched to specific diagnostic criteria will be needed.

Effect of Differences in Immunological Reconstitution By Donor Age and Recipient Age on the Incidence of Chronic Graft-Versus-Host Disease

Background: Development of chronic graft-versus-host disease (cGVHD) has been reported to be associated with donor age and recipient age. We hypothesized that differences in post-transplantation immunological reconstitution depending on donor age and recipient age might be responsible for the incidence of cGVHD. Redevelopment of regulatory T (Treg) cell expansion and cytokine production was investigated over time according to donor age and recipient age at the time of transplantation after allogeneic hematopoietic cell transplantation (HCT). Methods: The subjects were 39 consecutive patients who underwent HCT for different hematologic diseases at our hospital between December 2008 and December 2021 and survived for more than 3 years after HCT without cGVHD. The patients were classified into younger (age ≤59, n=23) and older (age ≥60, n=16) groups. The donor source was divided into subgroups based on use of cord blood or an adult donor: cord blood (younger, n=5; older, n=8) and adult donor (younger, n=18; older, n=8). Peripheral blood was drawn from all patients on day 200 and at 24 and 36 months after HCT. Analysis of Treg, T, B, and NK cells was performed using flow cytometry. T cells were isolated using EasySep kits and cultured at 37℃ in 5% CO 2 in air for 4 days with 10% heat-inactivated fetal calf serum as controls, interleukin-2 (IL-2) and transforming growth factor-β1 (TGFβ1) as samples, or concanavalin A (ConA) (10 µg/ml) at a density of 1×10 6 cells/ml. Treg cells were analyzed by flow cytometry and ConA-stimulated culture supernatants were tested for concentrations of IL-4, IL-5, IL-6, IL-10, IL-13, TGFβ1, tumor necrosis factor α (TNFα) and interferon-γ (IFNγ). Results: The results for patients who underwent HCT at our institution during the same observation period as this study suggested that donor source and patient age may be involved in development of cGVHD (p=0.032, p=0.015, respectively). In younger patients, CD4/8 in peripheral blood showed a tendency to increase over time after transplantation: 0.91±1.0 at 200 days, 0.54±0.29 at 2 years, and 1.32±0.57 at 3 years (R=0.308, p=0.072); and young patients after cord blood transplantation showed a marked improvement: 0.30±0 at 200 days, 0.59±0.18 at 2 years, and 1.84±0.27 at 3 years (R=0.913, p=0.0016). Percentages of Treg cells in peripheral blood and stimulated cultures were positively correlated in younger (R=0.349, p=0.043) and older (R=0.381, p=0.029) patients. The percentage, absolute number, and amplification rate of Treg cells in stimulation culture of T cells from younger patients (6.37±4.02%, 3759.4±1963.3 /mL, 12.4±11.5 times) at 200 days after transplantation were significantly higher than those for older patients (3.04±1.48%, 1783.9±1152.0 /mL, 5.4±3.7 times) (p=0.027, p=0.003, p=0.023). Post-transplant Treg cell expansion rates in younger cord blood transplant recipients significantly increased by 2.4±0 times at 200 days, 2.8±0.6 times at 2 years, and 5.8±2.1 times at 3 years (R=0.711, p=0.046). Regarding cytokine production, IL-5 in stimulation culture from younger patients showed a tendency to increase over time: 4.75±8.89 pg/mL at 200 days, 20.04±27.20 pg/mL at 2 years, and 59.51±104.93 pg/mL at 3 years (R=0.294, p=0.087). Conclusions: T cell reconstitution and cytokine production tended to increase over time after transplantation in younger patients. The ability to reconstitute Treg cells was higher in younger patients early after transplantation and increased over time in young cord blood transplant recipients. These factors may account for the high incidence of cGVHD in older transplant recipients from adult donors, although the number of cases in the study is limited and the data are preliminary.

Myeloid Dendritic Cells Were Expanded and Functionally Stronger in Aplastic Anemia

At present, the activated T cell-mediated destruction of hematopoietic stem/progenitor cells is the widely-recognized pathogenesis of aplastic anemia (AA). As the most important antigen-presenting cells in humans, dendritic cells (DCs) play an important role in the upstream link of immune pathogenesis in AA patients, so it may be the reason for the breakdown of immune tolerance in AA patients. In addition, abatacept, a cytotoxic T lymphocyte-associated antigen-4 (CTLA-4) fusion protein, can block T cell activation and function by binding CD80 and CD86 and blocking their interaction with CD28. Thus may be a potential direction for targeting DCs to treat AA. However, relevant reports have not been seen so far. To investigate the potential role of DCs in the pathogenesis of AA, we collected peripheral blood (PB) and bone marrow (BM) and measured the proportion of DCs. Results revealed that compared with healthy donors (HD), the proportion of myeloid DC in PB and BM in AA patients was significantly higher. In contrast, the proportion of plasmacytoid DC decreased significantly. Besides, the expression of CD80 and CD86 on PB DC surface of AA patients was higher than that of HD patients. Next, we cultured monocyte-derived DC in vitro and detected its phenotype and function. We confirmed that compared with HD, the expression rates of the co-stimulatory molecule CD80 and mature molecule CD83 of DC in AA patients were significantly increased, the apoptosis level was significantly decreased, and the ability to promote proliferation of CD3 +, CD4 + and CD8 + T lymphocytes was enhanced. At the same time, AA-DC significantly enhanced the ability of CD4 + T and CD8 + T cells to express CD25 and CD69. In addition, the ability of AA-DC to promote the differentiation of Th0 cells to Th1 and Th17 cells was stronger than HD. The ability to promote the differentiation of Th0 to Treg cells was weaker than HD. To investigate the feasibility of abatacept targeting DC in the treatment of AA, we compared the proportions of CTLA-4 and found the expression of CTLA-4 on CD4 + T cells and Treg cells in PB of AA patients was significantly decreased. Then, our in vitro tests confirmed that abatacept can bind to CD80 on the surface of DC and significantly down-regulate the expression of CD80. Adding abatacept into the co-culture system of DC and CD3 + T lymphocytes significantly inhibited the proliferation of CD3 +, CD4 + and CD8 + T lymphocytes, and significantly inhibited the expression of CD25 and CD69 in CD4 + T and CD8 + T cells. In addition, the ability of DC to promote the secretion of interferon-γ by CD4 + and CD8 + T cells in AA patients was significantly weakened after the addition of abatacept, while the ability to promote the secretion of interleukin-4 by CD4 + and CD8 + T cells was significantly enhanced. Finally, we performed RNA-Seq analysis using DCs cultured in vitro from HD and AA patients and using DC from AA patients added with abatacept to uncover the possible molecular mechanism. Result showed that compared with HD, there were significant differences in the expression levels of 58 genes of DC in patients with AA, including 22 up-regulated genes and 36 down-regulated genes. Go analysis inferred that the differentially expressed genes were mainly concentrated in cell migration, cell chemotaxis, regulation of cytokine production, and immune response. Besides, KEGG pathway analysis implied that the signaling pathway of significant enrichment of differentially expressed genes was cytokine-cytokine receptor interaction, NF-κB signaling pathway, chemokine signaling pathway and IL-17 signaling pathway. Meanwhile, compared with AA patients, after adding abatacept, 1058 genes showed significant differences in expression levels. Go analysis inferred that the differentially expressed genes were mainly concentrated in immune response, cytokine secretion, regulation of hemopoiesis, cell activation, differentiation, proliferation, and adhesion. Moreover, KEGG pathway analysis implied that the signaling pathway enriched in cytokine-cytokine receptor interaction, JAK-STAT signaling pathway, NF-κB signaling pathway and Th1, Th2 and Th17 cell differentiation. Our finding revealed the increased number and hyperfunction of DC in AA patients may be involved in the immune pathogenesis of AA. Abatacept can block T cell activation by blocking the DC surface co-stimulatory molecule CD80 and is a promising strategy for AA treatment.

CD8+ Treg cells play a role in the obesity-associated insulin resistance

Obesity-related chronic low-grade inflammation may trigger insulin resistance and type 2 diabetes (T2D) development. Cells with regulatory phenotype have been shown to be reduced during obesity, especially CD4+ Treg cells. However, little is known about the CD8+ Treg cells. Therefore, we aim to characterize the CD8+ Treg cells in human peripheral blood and adipose tissue, specifically, to address the effect of obesity and insulin resistance in this regulatory immune cell population. A group of 42 participants with obesity (OB group) were recruited. Fourteen of them were evaluated pre- and post-bariatric surgery. A group of age- and sex-matched healthy volunteers (n = 12) was also recruited (nOB group). CD8+ Treg cell quantification and phenotype were evaluated by flow cytometry, in peripheral blood (PB), subcutaneous (SAT), and visceral adipose tissues (VAT). The OB group displayed a higher percentage of CD8+ Treg cells in PB, compared to the nOB. In addition, they were preferentially polarized into Tc1- and Tc1/17-like CD8+ Treg cells, compared to nOB. Moreover, SAT displayed the highest content of CD8+ Tregs infiltrated, compared to PB or VAT, while CD8+ Tregs infiltrating VAT displayed a higher percentage of cells with Tc1-like phenotype. Participants with pre-diabetes displayed a reduced percentage of TIM-3+CD8+ Tregs in circulation, and PD-1+CD8+ Tregs infiltrated in the VAT. An increase in the percentage of circulating Tc1-like CD8+ Treg cells expressing PD-1 was observed post-surgery.In conclusion, obesity induces significant alterations in CD8+ Treg cells, affecting their percentage and phenotype, as well as the expression of important immune regulatory molecules.

The expression of Th17/Treg in oral submucosal fibrosis carcinogenesis and the significance in the development of mucosal lesions.

It targets to explore the expression of Th17/Treg in oral submucous fibrosis (OSF) carcinogenesis and its significance in the development of mucosal lesions. In this research, 100 patients with OSF who visited our hospital for surgical treatment from March 2020 to April 2022 were selected. Based on pathological examination results, the patients were divided into 27 patients with oral leukoplakia (OK) group, 14 patients with oral lichen planus (OLP) group, 9 patients with oral squamous cell carcinoma (OSCC) group, and 50 patients with OSF group. It adopted flow cytometry (FC) to calculate the ratio of peripheral blood Th17 cells and Treg cells in four groups, and the Th17/Treg ratio was calculated; The area of oral mucosal lesions (OML) from patients was collected. It needs to compare the differences in Th17/Treg ratio and OML area among four groups and determine the correlation between indicators. ROC curve was used to analyze the diagnostic threshold of the Th17/Treg ratio for carcinogenesis. Except for the OK and OLP, it had statistical significance differences in Th17, Treg cells, and Th17/Treg ratio (P<0.001); The area of OML in the OK, OLP, and OSCC was higher than that in the simple OSF, with statistical significance (P<0.001); Th17 (%), Treg (%), and Th17/Treg all had direct ratio with the area of OML; The area of OML has directed ratio with the development of mucosal lesions (r>0, P<0.05); The areas under the ROC curve for patients with OSF combined with OK, OLP, or OSCC with Th17 (%), Treg (%), Th17/Treg, and OML area were 0.560, 0.986, 0.936, and 0.466, respectively. The expression of Th17/Treg is elevated in oral submucosal fibrosis and carcinogenesis. When mucosal lesions progress or become cancerous, the Th17/Treg ratio increases accordingly, and it has more clinical value than the increase in the OML area.

The Effect of Dose-Fractionation Mode on the Immune Response and Therapeutic Effect of Lewis Lung Cancer Mice Model Based on Tumor Microenvironment

The therapeutic effect of different groups of Lewis lung cancer mice and the expression of immune cells in peripheral blood and tumor tissues of mice were observed after different dose-segmentation modes of treatment, so as to select the most suitable for stimulating immune effect with good prognosis and small side effects. Female homologous inbred C57BL/6 male mice at 6-8 weeks were selected and inoculated subcutaneously on the right thigh according to LLC: L929: lymphocyte = tumor cell suspension with PBMC(inactivated lymphocyte) = 5:1:1 ratio; after tumor growth to the size of 100-200mm3, different dose-segmentation modes were used for irradiation, which was divided into 5 groups, group A: no radiotherapy; Group B: 2Gy/qd, continuous radiotherapy for 30 days; Group C: 2Gy/bid, continuous radiotherapy for 13 days; Group D: 3Gy/bid, continuous radiotherapy for 10 days; Group E: 6Gy/qd, continuous radiotherapy for 8 days. On the 3rd day after radiotherapy, some mice were sacrificed, peripheral blood lymphocytes of mice were isolated with lymphocyte separation solution, and the transplanted tumor of mice was prepared into single-cell suspension. Then, the number of CD3+T cells, CD3+B cells, NK1.1 cells, CD3+CD4+T cells, CD3+CD8+T cells, CD4+Foxp3+Treg cells in peripheral blood and single-cell suspension of mice was detected by flow cytometry. HE staining was used to observe the tumor tissue structure of mice leg transplantation. The expressions of CD4, CD8, Foxp3 and CD31 in tumor tissues of each radiotherapy group were observed by immunohistochemical staining (IHC) in paraffin sections. The survival time of mice in group C (2Gy/ bidx13d) was significantly improved compared with that in group B (2Gy/qdx30d), and the twin-daily segmentation mode could improve the elevation of Treg cells after once-daily segmentation mode radiotherapy in both peripheral blood and tumor tissues. There was no significant difference in the survival time of mice in group E (6Gy/qdx8d) and group D (3Gy/bidx10d), the number of Treg cells in peripheral blood of mice in group E was significantly lower than that in group D, but there were more Treg cells in tumor tissue of mice in group E than group D. Compared with the control mice, the overall immunity of the experimental group was inhibited. The twice-daily segmentation mode in the conventional dose radiotherapy group significantly improved Treg in peripheral blood and tumor immune microenvironment and promoted neovascularization. The twice-daily segmentation mode in the high-dose radiotherapy group improved the immunosuppression of lymphocytes in peripheral blood but increased immunosuppression in tumor microenvironment.

Effect of sevoflurane on CD4+CD25+FOXP3+ regulatory T cells in patients with gastric cancer undergoing radical surgery.

In this study, the proportion of CD4+CD25+FOXP3+ regulatory T (Treg) cells in CD4+ T cells in the peripheral blood of gastric cancer patients before anesthesia induction (T1), after surgery (T2) and the first day after surgery (T3) was studied to explore the effect of sevoflurane and propofol anesthesia on the prognosis of gastric cancer patients. Forty patients with advanced gastric cancer were recruited and randomly divided into the sevoflurane group (S group) and the propofol group (T group). Flow cytometry was used to detect the proportion of CD4+CD25+FOXP3+ Treg cells in CD4+ T cells in the peripheral blood of patients with T1, T2 and T3, respectively. Compared with stage ⅡB, the proportion of CD4+CD25+FOXP3+ Treg cells in T1, T2 and T3 of stage ⅢA and stage ⅢB patients was increased. Compared with the T group, the expression of CD4+CD25+FOXP3+ Treg cells in the peripheral blood of T2 and T3 in the S group was decreased. The results showed that the expression of CD4+CD25+FOXP3+ Treg cells might be related to the TNM stage of gastric cancer and sevoflurane could alleviate the inhibition of postoperative immune function more than propofol. Sevoflurane effectively reduced the expression level of CD4+CD25+FOXP3+ Treg cells in peripheral blood of T2 and T3 of patients with gastric cancer, providing the theoretical basis for the selection of surgical anesthetics for patients with gastric cancer.

Defective Treg generation and increased type 3 immune response in leukocyte adhesion deficiency 1

In 15 Turkish LAD-1 patients and controls, we assessed the impact of pathogenic ITGB2 mutations on Th17/Treg differentiation and functions, and innate lymphoid cell (ILC) subsets.The percentage of peripheral blood Treg cells, in vitro-generated induced Tregs differentiated from naive CD4+ T cells were decreased despite the elevated absolute counts of CD4+ cells in LAD-1 patients. Serum IL-23 levels were elevated in LAD-1 patients. Post-curdlan stimulation, LAD-1 patient-derived PBMCs produced more IL-17A. Additionally, the percentages of CD18-deficient Th17 cells expanded from total or naïve CD4+ T cells were higher. The blood ILC3 subset was significantly elevated in LAD-1. Finally, LAD-1 PBMCs showed defects in trans-well migration and proliferation and were more resistant to apoptosis.Defects in de novo generation of Tregs from CD18-deficient naïve T cells and elevated Th17s, and ILC3s in LAD-1 patients' peripheral blood suggest a type 3-skewed immunity and may contribute to LAD-1-associated autoimmune symptoms.

IL-8 in bone marrow and peripheral blood of patients with B-cell acute lymphoblastic leukemia is associated with high regulatory T cell counts, degree of tumor infiltration and expression of CXCR1 in blasts

IntroductionRegulatory T cells (Treg cells) in a tumor environment and the expression of forkhead box P3 (FOXP3) in tumor cells have been associated with a poor prognosis. There are few studies evaluating Treg cells and FOXP3 in B-cell acute lymphoblastic leukemia (B-cell ALL). This study aimed to evaluate the frequencies of Treg cells in bone marrow (BM) and peripheral blood (PB) of patients with B-cell ALL and to determine their associations with the circulating cytokine profile and the expression of CXCR1 (IL-8 receptor) in Treg cells, as well as to compare FOXP3 expression in blasts of patients with B-cell ALL and normal lymphoid precursors. MethodsSamples of BM and PB from patients with B-cell ALL and healthy controls were studied. Treg cells, cytokines, FOXP3 and CXCR1 were evaluated using flow cytometry and analyzed. ResultsA total of 20 patients with B-cell ALL and 10 healthy controls were included. In B-cell ALL patients, Treg cell frequencies increased significantly, with higher percentages in the PB. Absolute Treg cell counts were associated with absolute blast counts in the BM and PB and with an IL-8 concentration. The IL-8 and IL-6 levels were associated with the CXCR1 expression in PB Treg cells. In addition, a greater expression of FOXP3 was observed in leukemic blasts than in normal lymphoid precursors. ConclusionsThese results suggest that the presence of Treg cells and cytokines in the tumor environment may correspond to mechanisms to evade the immune response. For that reason, it would be important to monitor these parameters in B-cell ALL to establish their effect on the disease prognosis.

The decreased of peripheral blood natural killer cell is associated with serum IL-2 level in the renal tubular acidosis in patients with primary sjogren’s syndrome

BackgroundPrimary Sjogren’s Syndrome (pSS) is a lymphoproliferative disease with autoimmune characteristics, which is characterized by lymphocyte infiltration of exocrine glands and involvement and dysfunction of extraglandular organs. Renal tubular acidosis (RTA) is a common renal involvement in pSS. This study investigated the phenotypic characteristics of peripheral blood lymphocyte subsets and cytokines in pSS patients complicated with RTA (pSS-RTA).MethodThis retrospective study included 25 pSS patients complicated with RTA and 54 pSS patients without RTA (pSS-no-RTA). To examine the level of peripheral lymphocytes subsets, flow cytometry analysis was used. The level of serum cytokines were detected by flow cytometry bead array(CBA). The influencing factors related to the occurrence of pSS-RTA were identified through logistic regression analyze.ResultsThe absolute number of CD4 + T cells and Th2 cells in peripheral blood were decreased in pSS-RTA patients than pSS-no-RTA patients. Moreover, the absolute number of NK cells and Treg cells were also decreased in pSS-RTA patients than pSS-no-RTA. The level of serum IL-2 was higher in pSS-RTA patients than pSS-no-RTA patients, and is negatively correlated with the number of NK cells, the number and percentage of Th17 cells, and Th17/Treg. Serum IL-2 level is also correlated with various cytokines. Multivariate logistic analysis proved that elevated ESR and ALP were risk factors for pSS complicated with RTA, while Treg was a protective factor.ConclusionThe increase of serum IL-2 level and the decrease of peripheral blood NK cells and Treg cells may be the immune mechanism of the development of pSS-RTA disease.

High doses of intravenous immunoglobulin stimulate regulatory T cell and suppress natural killer cell in women with recurrent pregnancy loss

The aim was to evaluate whether natural killer (NK) cells and regulatory T (Treg) cells were involved in mechanisms underlying beneficial effects of a high dose of intravenous immunoglobulin (IVIG) on recurrent pregnancy losses (RPL) of unexplained etiology. In a double-blind, randomized, placebo-controlled trial of IVIG (400 mg/kg, for 5 days in 4–6 weeks of gestation) in women with RPL, blood samples were collected pre-infusion, one week after infusion (1 w), and eight weeks of gestation/when miscarried (8 w). Levels of NK and Treg cells in peripheral blood were compared between women with IVIG (n = 50) and placebo (n = 49), and between women with IVIG who gave live birth (n = 29) and those who had miscarriage with normal chromosome (n = 12). Effector Treg cell percentages in IVIG group at 1 w (mean 1.43 % vs. 1.03 %) and at 8 w (1.91 % vs. 1.18 %) were higher than those in placebo group (p < 0.01). Total Treg cell percentages in IVIG group at 1 w (4.75 % vs. 4.08 %) and at 8 w (5.55 % vs. 4.47 %) were higher than those in placebo group (p < 0.05). In women with live birth, total Treg cell percentages increased at 8 w (5.52 %, p < 0.001) compared with pre-infusion (4.54 %) and 1 w (4.47 %), while NK cell activity decreased at 1 w (20.18 %, p < 0.001) compared with pre-infusion (26.59 %). IVIG increased Treg cell percentages and suppressed NK cell activity very early in pregnancy, and these were associated with subsequent live birth. Stimulation of Treg cells and suppression of NK cell activity very early in pregnancy may be a mechanism of pharmacological effects of high dose IVIG.

Integrative epigenetic analysis reveals AP-1 promotes activation of tumor-infiltrating regulatory T cells in HCC.

Regulatory T (Treg) cells that infiltrate human tumors exhibit stronger immunosuppressive activity compared to peripheral blood Treg cells (PBTRs), thus hindering the induction of effective antitumor immunity. Previous transcriptome studies have identified a set of genes that are conserved in tumor-infiltrating Treg cells (TITRs). However, epigenetic profiles of TITRs have not yet been completely deciphered. Here, we employed ATAC-seq and CUT&Tag assays to integrate transcriptome profiles and identify functional regulatory elements in TITRs. We observed a global difference in chromatin accessibility and enhancer landscapes between TITRs and PBTRs. We identified two types of active enhancer formation in TITRs. The H3K4me1-predetermined enhancers are poised to be activated in response to tumor microenvironmental stimuli. We found that AP-1 family motifs are enriched at the enhancer regions of TITRs. Finally, we validated that c-Jun binds at regulatory regions to regulate signature genes of TITRs and AP-1 is required for Treg cells activation in vitro. High c-Jun expression is correlated with poor survival in human HCC. Overall, our results provide insights into the mechanism of AP-1-mediated activation of TITRs and can hopefully be used to develop new therapeutic strategies targeting TITRs in liver cancer treatment.

Exenatide regulates Th17/Treg balance via PI3K/Akt/FoxO1 pathway in db/db mice

BackgroundThe T helper 17 (Th17)/T regulatory (Treg) cell imbalance is involved in the course of obesity and type 2 diabetes mellitus (T2DM). In the current study, the exact role of glucagon-like peptide-1 receptor agonist (GLP-1RA) exenatide on regulating the Th17/Treg balance and the underlying molecular mechanisms are investigated in obese diabetic mice model.MethodsMetabolic parameters were monitored in db/db mice treated with/without exenatide during 8-week study period. The frequencies of Th17 and Treg cells from peripheral blood and pancreas in db/db mice were assessed. The phosphoinositide 3-kinase (PI3K)/protein kinase B (Akt)/Forkhead box O1 (FoxO1) pathway in Th17 and Treg cells from the spleens of male C57BL/6J mice was detected by western blotting. In addition, the expression of glucagon-like peptide-1 receptor (GLP-1R) in peripheral blood mononuclear cells (PBMCs) of male C57BL/6J mice was analyzed.ResultsExenatide treatment improved β-cell function and insulitis in addition to glucose, insulin sensitivity and weight. Increased Th17 and decreased Treg cells in peripheral blood were present as diabetes progressed while exenatide corrected this imbalance. Progressive IL-17 + T cell infiltration of pancreatic islets was alleviated by exenatide intervention. In vitro study showed no significant difference in the level of GLP-1R expression in PBMCs between control and palmitate (PA) groups. In addition, PA could promote Th17 but suppress Treg differentiation along with down-regulating the phosphorylation of PI3K/Akt/FoxO1, which was reversed by exenatide intervention. FoxO1 inhibitor AS1842856 could abrogate all these effects of exenatide against lipid stress.ConclusionsExenatide could restore systemic Th17/Treg balance via regulating FoxO1 pathway with the progression of diabetes in db/db mice. The protection of pancreatic β-cell function may be partially mediated by inhibiting Th17 cell infiltration into pancreatic islets, and the resultant alleviation of islet inflammation.

Risk factors analysis and risk assessment model construction of systemic lupus erythematosus patients with infection.

To analyze the characteristics of peripheral blood lymphocyte subsets in systemic lupus erythematosus (SLE) patients with infection and non-infection group. Explore the risk factors of infection in SLE patients and establish a risk matrix model to predict the occurrence of co-infection. total of 333 SLE patients without infection, 163 patients suffering from infection, and 132 healthy controls (HCs) were recruited. General clinical data and disease activity indicators were collected. The levels of total T, B, CD4+T, CD8+T, NK, Th1, Th2, Th17, and Treg cells in peripheral blood of HCs, SLE patients (including infected and non-infected group) were analyzed by flow cytometry. The risk assessment model was constructed, and the receiver operating characteristic curve was drawn. 39 SLE patients with infection and 20 patients without infection were randomly selected to evaluate the predictive power of the regression model. The levels of T, B, CD4+T, CD8+T, and NK cells in the infected patients were significantly decreased when compared with that of both non-infected patients and HCs (p < .05). The non-infected patients had a higher level of Th17 than that of HCs (p < . 05), but the absolute numbers of Th17 in infected patients was the lowest among the three groups (p < .001). The number of Treg cells in SLE patients was significantly lower than that of HCs (p < .01), and the infected patients had the fewest Treg cells among all these groups (p < . 05). A risk assessment model for SLE with infection was established, p = 1/(1-e-y), Y = 1.763-0.004 × Absolute number of CD4 + T cells-0.005 × Absolute number of NK cells -0.005 × Platelet count(×1012/L) + 1.033 × Absolute number of lymphocytes (×109/L) + 0.023 × C-reactive protein (mg/dL), whose predictive sensitivity is 77.5%, and specificity is 78.3%. The new risk assessment model exhibits good predictive ability to assess co-infection risk in SLE patients. T cells, NK cells, and CD4 + T cells along with other parameters help in differentiating Lupus with infection from Lupus alone.

Different expression patterns of VISTA concurrent with PD-1, Tim-3, and TIGIT on T cell subsets in peripheral blood and bone marrow from patients with multiple myeloma.

V-type immunoglobulin domain-containing suppressor of T cell activation (VISTA) is considered as an immunosuppressive factor and potential therapeutic target for anticancer therapy. However, little is known about VISTA expression and its role in immunosuppression in multiple myeloma (MM). In this study, VISTA expression and co-expression with programmed cell death receptor-1 (PD-1), T cell immunoglobulin mucin-domain-containing-3 (Tim-3), and T cell immunoglobulin and ITIM domain (TIGIT) in CD3+, CD4+, CD8+, and regulatory T (Treg) cells were analyzed in patients with MM by multi-color fluorescent flow cytometry of peripheral blood (PB) and bone marrow (BM) samples from 36 patients with MM and compared to 36 PB samples and 10 BM samples from healthy individuals (HIs), which served as controls. The results demonstrated a significant increased percentage of VISTA co-expression with PD-1, Tim-3, and TIGIT in CD3+, CD4+, CD8+, and Treg cells in PB from MM patients compared with HIs. A similar trend for VISTA+CD8+ T cells was found in BM. Moreover, a trend of a high percentage on VISTA expression and co-expression in PB rather than BM was found. Furthermore, significant positive correlations existed for VISTA expression concurrent with PD-1, Tim-3, and TIGIT in T cell subsets and clinical indicators, including Revised International Staging System (R-ISS) staging of multiple myeloma, Eastern Cooperative Oncology Group (ECOG) score, and beta-2-microglobulin (β2-MG). In conclusion, higher VISTA expression concurrent with PD-1, Tim-3, and TIGIT on T cells, particularly in the PB of patients with MM, may result in T cell exhaustion and dysfunction and be closely associated with disease progression and clinical indicators. Thus, VISTA may be considered a potential target for reversing T cell exhaustion and improving T cell function in MM.

Peripheral Treg Levels and Transforming Growth Factor-β (TGFβ) in Patients with Psoriatic Arthritis: A Systematic Review Meta-Analysis.

Studies on the level of regulatory T(Treg) cells in psoriatic arthritis (PsA) have been controversial, leading to disagreement regarding the role Treg cells play in the pathogenesis of the disease. To clarify the status of Treg cells in patients with PsA, we performed a meta-analysis to determine the levels of Treg cells and serum Treg-associated cytokines in PsApatients. According to published data from PubMed, Web of Science, Embase, Clinical Trials.gov, MEDLINE, Web of Knowledge, Cochrane Library, and FDA.gov, we determined the Treg and Treg cytokine levels in patients with PsA. The effect estimates were pooled using a random-effects model. This meta-analysis included 12studies. Compared to healthy controls (HCs), the proportions of Treg cells had no significant difference in patients with PsA (based on standardized means[SMD] = - 1.038, 95% confidence intervals[CI] = - 2.165 to 0.089, p = 0.071). On the basis of subgroup analysis, patients with PsA had a lower percentage of CD4+ Treg cells (SMD = - 1.501, 95%CI - 2.799 to - 0.202, p = 0.023) than OKT8+ Treg (SMD = 0.568, 95%CI - 2.127 to 3.263, p = 0.679). Besides, CD4+CD25+FoxP3+ Treg cells and CD4+CD25highCD127low Treg cells were both significantly decreased on the levels of PBMCs in patients with PsA (SMD = - 0.764, 95%CI - 1.404 to - 0.125, p = 0.019; SMD = - 5.184, 95%CI - 6.955 to - 3.412, p < 0.001). CD4+CD25+FoxP3+ Treg cells were particularly more abundant in the synovial fluid thanin peripheral blood (SMD = 3.288, 95%CI 2.127 to 4.449, p < 0.001). No significant difference was observed in the proportion of CD4+CD25+ Treg cells in peripheral blood and CD4+CD25+FoxP3+ Treg cells in CD4+ Tcells (SMD = - 2.498, 95%CI - 7.720 to 2.725, p = 0.349; SMD = - 0.719, 95%CI - 2.525 to 1.086, p = 0.435). PsA patients had decreased cytokines such as transforming growth factor-β (TGFβ) (SMD = - 2.199, 95%CI - 3.650 to - 0.749, p = 0.003). Treg definition markers influence the scale of Treg cells in patients with PsA. Pathogenesis of PsA may be attributed to an insufficient or malfunctioning Treg population.