scholarly journals Reduced Numbers and Impaired Function of Regulatory T Cells in Peripheral Blood of Ischemic Stroke Patients

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Johanna Ruhnau ◽  
Juliane Schulze ◽  
Bettina von Sarnowski ◽  
Marie Heinrich ◽  
Sönke Langner ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Peripheral Blood ◽  
Experimental Stroke ◽  
Stroke Patients ◽  
Effector Cells ◽  
T Effector Cells ◽  
Treg Function ◽  
Age Dependent ◽  
And Function

Background and Purpose. Regulatory T cells (Tregs) have been suggested to modulate stroke-induced immune responses. However, analyses of Tregs in patients and in experimental stroke have yielded contradictory findings. We performed the current study to assess the regulation and function of Tregs in peripheral blood of stroke patients. Age dependent expression of CD39 on Tregs was quantified in mice and men. Methods. Total FoxP3+ Tregs and CD39+FoxP3+ Tregs were quantified by flow cytometry in controls and stroke patients on admission and on days 1, 3, 5, and 7 thereafter. Treg function was assessed by quantifying the inhibition of activation-induced expression of CD69 and CD154 on T effector cells (Teffs). Results. Total Tregs accounted for 5.0% of CD4+ T cells in controls and <2.8% in stroke patients on admission. They remained below control values until day 7. CD39+ Tregs were most strongly reduced in stroke patients. On day 3 the Treg-mediated inhibition of CD154 upregulation on CD4+ Teff was impaired in stroke patients. CD39 expression on Treg increased with age in peripheral blood of mice and men. Conclusion. We demonstrate a loss of active FoxP3+CD39+ Tregs from stroke patient’s peripheral blood. The suppressive Treg function of remaining Tregs is impaired after stroke.

Selective Expansion of Regulatory T-Cells during Lenalidomid Treatment of MDS with Isolated Deletion 5q

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 4654-4654
Author(s):  
Martin Wermke ◽  
Claudia Schuster ◽  
Claudia Schönefeldt ◽  
Sebastian Tuve ◽  
Gerhard Ehninger ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Transforming Growth Factor Beta ◽  
Peripheral Blood ◽  
Th17 Cells ◽  
Transforming Growth Factor ◽  
Absolute Lymphocyte Count ◽  
Intracellular Staining ◽  
Effector Cells ◽  
T Effector Cells

Abstract Background Lenalidomide (LEN) results in hematopoietic improvement in the majority of treated patients with myelodysplastic syndrome and isolated deletion of the long arm of chromosome 5 (MDS-del5q). The drug is believed to exert antiangiogenetic, antiproliferative, proerythropoietic and immunomodulatory effects. The precise nature especially of the latter mode of action is incompletely understood. While some reports in multiple myeloma argue for an immunosuppressive effect, others proposed immunostimulatory properties. This prompted us to study the effects of LEN on the adaptive immune system in a cohort of patients with MDS-del5q. Patients, Material and Methods The composition of the T-cell compartment in peripheral blood (pB) and bone marrow (BM) of 11 patients with MDS-del5q treated within a multicenter, single-arm, phase II study of the German MDS study group (LEMON-5 trial) was studied by flow cytometry. Patients received LEN 10 mg per day for 21 days of each 28 day cycle. The median age was 71 years and 9 patients were female. Regulatory T-cells (Treg) were defined as CD3+CD4+CD25++CD127dimFoxP3+ while Th17 cells had to be CD3+CD4+ with positive intracellular staining for IL17 and negative intracellular staining for IFNgamma. Peripheral blood samples were taken prior to initiation of each new LEN cycle and BM specimens were obtained prior to treatment initiation and after 5 to 7 cycles of LEN. The results obtained in MDS patients were compared to a cohort of 27 healthy volunteer donors. Results After a median of 6 cycles all but one patient had responded with a hematologic improvement of erythrocytes, while 6 individuals achieved a complete morphologic and 5 a complete cytogenetic remission. In comparison to healthy donors, MDS patients had significantly lower frequencies of Treg in pB (5.0 vs. 7.4 %, p = 0.008) and BM (3.4 vs. 7.4 %, p = 0.006). After initiation of LEN treatment we observed a rapid increase in Treg frequencies in pB (Figure 1A) and after a median of 6 cycles the Treg were significantly more abundant in pB (9.6 % p = 0.001, Figure 1B) and BM (8.1 %, p = 0.016, Figure 1C) compared to baseline. The median Treg frequency reached the level observed in normal donors in BM and surpassed it in pB (Figure 1B+C). Fluorescence-in-situ hybridization demonstrated that the expanded Treg were not part of the malignant clone. Interestingly, although the median absolute lymphocyte count increased during LEN treatment (pre: 1.10 Gpt/l vs. post: 1.43 Gpt/l p = 0.013) there were no significant changes with respect to relative lymphocyte frequency (pre: 68.3 %, post: 75.61 %, p=0.499) or the percentage of T-effector cells (pre: 48.0 %, post: 51.5 %, p=0.520). Moreover, the expansion of Treg was not accompanied by a counter-regulatory increase of pro-inflammatory Th17-cells (pre: 2.3 % vs. post: 1.33 %, p = 0.328). To address a possible mechanism of the observed Treg expansion we analyzed the concentration of transforming growth factor beta (TGFbeta), a key factor contributing to the development of Treg, in pB and BM plasma. However, LEN treatment was not associated with an increase of TGFbeta levels in these two compartments. On the contrary, the TGFbeta levels after 6 cycles of LEN were significantly lower than pre-treatment values in pB and BM (pB 4.9 vs. 2.3 ng/ml, p = 0.039; BM 4.5 vs. 0.8 ng/ml, p = 0.023). Discussion Our results show that LEN treatment corrects an MDS-del5q-inherent Treg deficit. This Treg expansion is not accompanied by a counter-regulatory increase in pro-inflammatory Th17 or other T-effector cells and therefore possibly results in an immunosuppressive milieu, which might contribute to the beneficial effects of the drug on myelodysplastic hematopoiesis but may also facilitate leukemic progression. Figure 1 Figure 1. Disclosures Platzbecker: Celgene: Research Funding.

Decreased LAG3 expression on T effector cells and regulatory T cells in SAA

2020 ◽  
Vol 112 (6) ◽  
pp. 757-763
Author(s):  
Yingying Sun ◽  
Chunyan Liu ◽  
Ting Jiao ◽  
Ning Xie ◽  
Huaquan Wang ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Effector Cells ◽  
T Effector Cells

Quantitative and Qualitative Analysis Of Regulatory T Cells (Tregs) Derived From The Peripheral Blood Of Chronic Lymphocytic Leukemia Patients

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 5280-5280
Author(s):  
Eleni Dikaia Ioannidou ◽  
Vassiliki Mpakou ◽  
Myrofora Vikentiou ◽  
Eugenia Konsta ◽  
Frieda Kontsioti ◽  
...  
Keyword(s):  
T Cells ◽  
B Cell ◽  
Peripheral Blood ◽  
Mononuclear Cells ◽  
Annexin V ◽  
Treg Cells ◽  
Lymphocytic Leukemia ◽  
Effector Cells ◽  
T Effector Cells ◽  
Healthy Donors

Abstract Introduction T regulatory cells are immunosuppressive cells, which are considered to play an important role in the regulation of immune response to cancer, by restraining autoreactive lymphocytes. Several studies, mostly in solid tumors, revealed that the number of Treg cells increases as the disease progresses and that Treg cells act by suppressing anti-tumor immune response, through the targeting of other immune cells, such as T cells, B cells and dendritic cells. Chronic lymphocytic leukemia (CLL) is a lymphoid malignancy, characterized by both, immunodeficiency and autoimmune disorders. Accumulated data indicate the role of T cells in the pathogenesis and development of CLL and reveal an increased number of Treg cells in CLL patients. The scope of this study is the analysis of the functional role of Tregs derived from the peripheral blood of CLL patients, mainly on B-CLL cells, and its correlation with well known prognostic factors. Methods Treg cells derived from mononuclear cells of 28 untreated B-cell CLL patients with a median age 62 (44-88) and 17 healthy donors were analyzed through Flow cytometry. Patients were classified according to Rai classification as Rai I:19, Rai II:4, Rai III:5 and according to Binet as Binet A: 24, Binet B:3 and Binet C:1. The following antibodies were used for the fluorescence-activated cell sorter (FACS) analysis: 1. CD45Ro-FITC/CD45RA-PE/CD4-ECD/CD25-PC5/CD127-PC7 2. CD1a-FITC/CD137-PE/CD4-ECD/CD25-PC5/CD127-PC7 3. CD95-FITC/cyCD152-PE/CD4-ECD/CD25-PC5/CD127-PC7 4. beads/FoxP3-PE/CD4-ECD/CD25-PC5/CD127-PC7 5. Annexin V-FITC/CD4-ECD/CD25-PC5/CD127-PC7 Moreover, peripheral blood was obtained from 15 patients with B-cell CLL. Mononuclear cells were isolated using Ficoll-Paque gradient centrifugation. CD4+CD25+ (Treg cells), CD4+CD25- (T effectοr cells, Teff), CD5+CD19+ (B-CLL) and CD5+CD19- (Normal B, NB) cells were separated using magnetic antibody cell sorting. To test the functionality of the assayed Tregs, the isolated cell populations were cultured in a 96-well plate (Tregs, Teff, B-CLL, NB cells, Tregs:Teff in a 4:1 ratio, B-cll:Tregs in 1:20 ratio, B-cll:Teff in 1:20 ratio, NB cells:Tregs in 1:20 ratio, NB cells:Teff in 1:20 ratio) and their proliferative capacity was measured using the BrdU assay. Results FACS analysis of the Treg cells resulted at the following observations: (1) The co-expression of the CD45RA-CD45RO markers was significantly higher in patients’ samples than in controls (p=0.047). (2) No significant differences were observed between patients and controls, regarding the expression of the CD1α marker, as well as the expression of CD95 and CD152 markers. (3) The Treg absolute cell number (cells/μL), estimated either as the number of CD4+ CD25+ CD127- cells or as the number of CD4+ CD25+ FoxP3+ cells, was statistically significantly higher in patients’ samples than in controls (CD127- p=0.047, FoxP3+ p= 0.036). (4) Annexin V expression in Treg cells from B- CLL patients was significantly lower compared to controls (p=0.027). Following the purification and culturing of T and B cells from B-cell CLL patients’ samples, functional analysis of the different cell populations was performed using the BrdU proliferation assay. We observed that Tregs were able to significantly suppress the proliferation of the Teff cells (p=0.002). After the co-culturing of NB cells (CD5+CD19-)and Tregs (CD4+CD25+) we found that NB cells seemed to significantly increase the proliferation of Treg cells, compared to the proliferation capacity of the Tregs when cultured alone (p=0.047). Moreover, we observed that Teff (CD4+CD25-) were able to significantly suppress the proliferation of B-CLL cells (CD5+CD19+), when co-cultured (B-CLL: Teff, 1:20 ratio) (p=0.05). Conclusions In B-cell CLL patients, Treg cells are significantly higher and present with lower apoptotic levels compared to healthy donors’ samples. The functional analysis of Treg cells indicates that they can effectively suppress the proliferation of T effector cells. Moreover, T effector cells seem to suppress the proliferation of B-CLL cells, while NB cells increase the proliferation of Treg cells. These observations could probably indicate that at the early stages of the disease, where NB cells are more aberrant, Treg cells’ activity is induced, leading to Teff cells’ suppression and therefore, to an indirect induction of B-CLL cells’ proliferation. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Regulatory T cell expansion by a highly CD25-dependent IL-2 mutein arrests ongoing autoimmunity

2019 ◽  
Author(s):  
Liliane Khoryati ◽  
Minh Nguyet Pham ◽  
McKenna Sherve ◽  
Swarnima Kumari ◽  
Kevin Cook ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Dose Range ◽  
Interleukin 2 ◽  
Autoimmune Response ◽  
Wild Type ◽  
Effector Cells ◽  
Inflammatory Conditions ◽  
Receptor Component ◽  
And Function

AbstractInterleukin-2 (IL-2) controls the homeostasis and function of regulatory T cells (Tregs) and defects in the IL-2 pathway contribute to multiple autoimmune diseases. Although recombinant IL-2 therapy has been efficacious in certain inflammatory conditions, the capacity for IL-2 to also activate inflammatory effector responses highlights the need for IL-2-based therapeutics with improved Treg-specificity. From a panel of rationally designed IL-2 variants, we identified IL-2 muteins with reduced potency and enhanced Treg-selectivity due to increased dependence on the IL-2-receptor component CD25. As an Fc-fused homodimer, the optimal Fc.IL-2 mutein induced selective Treg enrichment and reduced agonism of effector cells across a wide dose range. Furthermore, despite being a weaker agonist, overall Treg growth was greater and more sustained due to reduced receptor-mediated clearance of the Fc.IL-2 mutein compared to Fc-fused wild-type IL-2. Preferential Treg enrichment was also observed in the presence of activated pathogenic T cells in the autoimmune target organ, despite a loss of Treg-selectivity in an IL-2R-proximal response. These features allowed for extended resolution of spontaneous autoimmunity using infrequent dosing schedules. Thus, IL-2 muteins enable efficient, flexible, and targeted control of the autoimmune response.One Sentence SummaryA CD25-dependent IL-2 mutein selectively expands regulatory T cells and provides potent and targeted control of autoimmunity.

Abnormal quantity and function of regulatory T cells in peripheral blood of patients with severe aplastic anemia

2015 ◽  
Vol 296 (2) ◽  
pp. 95-105 ◽  
Author(s):  
Li Yan ◽  
Rong Fu ◽  
Hui Liu ◽  
Huaquan Wang ◽  
Chunyan Liu ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Aplastic Anemia ◽  
Peripheral Blood ◽  
Severe Aplastic Anemia ◽  
And Function

JAK Inhibition Reduces CD25 high CD27+ FOXp3+ T Regulatory Cells and Causes a Silencing Of T Effector Cells In Patients With Myeloproliferative Neoplasms Whilst Promoting a TH17 Phenotype

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 4092-4092 ◽  
Author(s):  
Clodagh Keohane ◽  
Shahram Y Kordasti ◽  
Thomas Seidl ◽  
Pilar Perez Abellan ◽  
Nicholas Shaun B. Thomas ◽  
...  
Keyword(s):  
T Cells ◽  
Inflammatory Cytokines ◽  
Peripheral Blood ◽  
T Regulatory Cells ◽  
Myeloproliferative Neoplasms ◽  
Healthy Controls ◽  
Regulatory Cells ◽  
Effector Cells ◽  
T Effector Cells

Abstract Background The myeloproliferative neoplasms (MPN), in particular myelofibrosis, are associated with elevated levels of inflammatory cytokines and constitutional symptoms. Treatment with JAK inhibitors (JAKi) have lead to marked improvement in symptoms and splenomegaly. Signaling through the JAK pathway is critical for T cell development and differentiation. However the baseline immune signature remains largely undescribed in MPN as does the effect of JAK inhibition on the immune subsets in this disease. Materials and Methods The % and absolute number of CD4+ T cell subsets (TH1, TH2 and TH17 and Foxp3+ T regulatory cells) in peripheral blood (PB) were investigated by flow cytometry. T cells were stimulated and stained intracellularly for IFNg, IL-4, IL-17 & TNFα. Tregs were defined as CD4+ CD25highCD27+FOXp3+. The serum level of 30 cytokines was also measured by Luminex. Patients received either ruxolitinib (n=21) or SAR302503 (n=13) as JAKi. Results We analysed 50 MPN patients (30 Myelofibrosis, 15 Polycythemia Vera, 5 Essential Thrombocythemia) and 14 healthy donors (HD). 34 patients were treated with JAKi and sequential PB samples were obtained at 1, 3, 6 and 12 month intervals (median follow up 6 months). Tregs are significantly lower in MPN patients compared to HD and drop further following treatment (p<0.0001 and p=0.0049 respectively). There was no difference at baseline in the T effector subsets between the groups including TH1, TH2 and TH17 secreting cells but there was a significant increase in TH17 following JAKi therapy (fig 1a). JAKi resulted in a significant decrease (p=0.03) in CD4 T cells secreting pro-inflammatory cytokines at 3 months follow up although this was less evident at 6 months follow up and occurred irrespective of disease response to treatment. This silencing was confirmed by both intracellular staining and luminex assay of supernatants including a significant decrease in Interleukin-2 receptor (IL-2r) p=0.0007, Interferon gamma induced protein (IP-10) p=0.0006, monokine induced by gamma interferon (MIG) p=0.0008 and hepatocyte growth factor (HGF) p=0.0009. This finding was reproduced in-vitro in healthy peripheral blood mononuclear cells (PBMCs). PBMCs were treated with the JAKi ruxolitinib (100-300NM) in the presence or absence of plate bound anti-CD3/28 stimulation and cultured for 5 days. Tregs were reduced in number and there was a considerable increase in the percentage of “cytokine negative” or “silent” T effector cells by FACS analysis compared to untreated or vehicle treated cells (median of 42 % of CD4 to 91% of CD4) (fig 1b). This finding was reproduced by Luminex cytokine assay of supernatants. Western blot demonstrated a reduction in pSTAT3 in ruxolitinib treated cells. To assess the effect of JAKi on Treg function, healthy isolated Tregs were treated with ruxolitinib and co cultured with CFSE labeled autologous T effector cells. Short term JAKi treated Tregs were unable to suppress the proliferation of T effector cells compared to Tregs treated with vehicle alone. Similarly, proliferation rate and function of Tregs was reduced following 4 weeks expansion in the presence of ruxolitinib compared to expanded Tregs in the presence of ATRA and rapamycin as a control. Conclusions Tregs are significantly lower in MPN patients compared to healthy controls in keeping with the inflammatory environment of MPN and decrease further with JAKi. Surprisingly, T effector numbers were not significantly different to healthy controls at baseline and TH1 and TH2 subsets did not change with therapy. However, secretion of proinflammatory cytokines from these cells was blocked with JAKi both invivo and invitro resulting in a functional silencing of T effectors. Interestingly, TH17 subsets increase with treatment possibly representing a polarization from a Treg phenotype to a TH17 phenotype, suggesting the re-establishment of immune-surveillance against the malignant clone. Further investigation is required to confirm the hypothesis that these expanded TH17 cells originate from Tregs or previously “silenced” CD4 T cells. Disclosures: Harrison: Novartis: Honoraria; Sanofi : Honoraria.

scholarly journals Induced, but not natural, regulatory T cells retain phenotype and function following exposure to inflamed synovial fibroblasts

2020 ◽  
Vol 6 (44) ◽  
pp. eabb0606 ◽  
Author(s):  
Sujuan Yang ◽  
Ximei Zhang ◽  
Jingrong Chen ◽  
Junlong Dang ◽  
Rongzhen Liang ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Synovial Fibroblasts ◽  
Foxp3 Expression ◽  
Regulatory Function ◽  
Migration And Invasion ◽  
Effector Cells ◽  
T Effector Cells

Aberrant number and/or dysfunction of CD4+Foxp3+ Regulatory T cells (Tregs) are associated with the pathogenesis of rheumatoid arthritis (RA). A previous study has demonstrated that thymus-derived, natural Tregs (nTregs) prefer to accumulate in inflamed joints and transdifferentiate to TH17 cells under the stimulation of inflamed synovial fibroblasts (SFs). In this study, we made a head-to-head comparison of both Treg subsets and demonstrated that induced Tregs (iTregs), but not nTregs, retained Foxp3 expression and regulatory function on T effector cells (Teffs) after being primed with inflamed SFs. In addition, iTregs inhibited proliferation, inflammatory cytokine production, migration, and invasion ability of collagen-induced arthritis (CIA)–SFs in vitro and in vivo. Moreover, we noted that iTregs directly targeted inflamed SFs to treat autoimmune arthritis, while nTregs failed to do this. Thus, manipulation of the iTreg subset may have a greater potential for prevention or treatment of patients with RA.

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