scholarly journals Features of immunoregulation in patients with pulmonary tuberculosis with blood eosinophilia

2018 ◽  
Vol 17 (3) ◽  
pp. 168-179
Author(s):  
O. I. Urazova ◽  
E. G. Churina ◽  
Yu. V. Kolobovnikova ◽  
V. V. Novitskiy ◽  
A. V. Karaulov ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Regulatory T Cells ◽  
Pulmonary Tuberculosis ◽  
Venous Blood ◽  
Enzyme Linked Immunosorbent Assay ◽  
Mononuclear Leukocytes ◽  
Blood Eosinophilia ◽  
Immune Response Regulation

The aimof the investigation was to determine the characteristics of the immune response regulation for pulmonary tuberculosis (TB) and to analyze the role of regulatory T cells in the immunopathogenesis of TB with eosinophilia in the blood, depending on the clinical form of the disease and sensitivity of Micobacterium tuberculosis to anti-TB drugs.Materials and methods.157 patients who were initially diagnosed with infiltrative and disseminated TB were examined. The material of the study was venous blood and culture of mononuclear leukocytes isolated from venous blood. The content of interleukin (IL) 4, IL-10 and transforming factor beta (TGFβ) in culture suspensions of mononuclear leukocytes in vitro and IL-5 in the blood was determined by enzyme-linked immunosorbent assay (ELISA) test. The expression of surface molecules CD4, CD20, CD25 and intracellular transcription factor Foxp3 by lymphocytes of the blood was evaluated by flow cytometry. The obtained results were analyzed by statistical methods.Results.It is shown that excessive generation of regulatory T cells in patients with TB is associated with eosinophilia of the blood and imbalance of immune response regulation mechanisms. In TB with eosinophilia, an increase in the number of Foxp3-positive regulatory T cells in the blood is combined with in vitro hypersecretion of anti-inflammatory cytokines TGFβ, IL-10, IL-4 and an increase in the content of CD20+ B lymphocytes and IL-5 in the blood. These changes are most pronounced in the disseminated form of TB in combination with drug resistance.Conclusion.Characteristics of immunoregulation at TB with blood eosinophilia are associated with activation of immunosuppression mechanisms and polarization of immune response towards Th2-dependent pathway.

scholarly journals T helper cell polarisation as a measure of the maturation of the immune response

2003 ◽  
Vol 12 (5) ◽  
pp. 285-292 ◽  
Author(s):  
Scott B. Cameron ◽  
Ellen H. Stolte ◽  
Anthony W. Chow ◽  
Huub F. J. Savelkoul
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Intracellular Cytokine Staining ◽  
T Helper Cell ◽  
Helper Cell ◽  
Enzyme Linked Immunosorbent Assay ◽  
Intracellular Cytokine ◽  
T Helper

Background:T helper cell polarisation is important under chronic immune stimulatory conditions and drives the type of the evolving immune response. Mice treated with superantigensin vivodisplay strong effects on Thsubset differentiation. The aim of the study was to detect the intrinsic capacity of T cells to polarise under variousex vivoconditions.Methods:Purified CD4+T cells obtained from superantigen-treated mice were cultured under Thpolarising conditionsin vitro. By combining intracellular cytokine staining and subsequent flow cytometric analysis with quantitative cytokine measurements in culture supernatants by enzyme-linked immunosorbent assay (ELISA), the differential Thpolarising capacity of the treatment can be detected in a qualitative and quantitative manner.Results and conclusions:BALB/c mice were shown to be biased to develop strong Th2 polarised immune responses using Th0 stimulation of purified CD4+T cells from phosphate-buffered saline-treated mice. Nevertheless, our analysis methodology convincingly showed that even in these mice, Toxic Shock Syndrome Toxin-1 treatmentin vivoresulted in a significantly stronger Th1 polarising effect than control treatment. Our results indicate that populations of Thcells can be assessed individually for their differential Th1 or Th2 maturation capacityin vivoby analysing robustin vitropolarisation cultures combined with intracellular cytokine staining and ELISA.

The in vitro effect of dexamethasone on maturation and differentiation of CD4CD45RO T-cells in rheumatoid arthritis

2017 ◽  
pp. 87-97
Author(s):  
Н.М. Тодосенко ◽  
К.А. Юрова ◽  
О.Г. Хазиахматова ◽  
И.П. Малинина ◽  
Л.С. Литвинова
Keyword(s):  
Rheumatoid Arthritis ◽  
T Cells ◽  
Negative Impact ◽  
Venous Blood ◽  
Cell Receptor ◽  
Mononuclear Leukocytes ◽  
Flow Cytofluorometry ◽  
Proinflammatory Mediators ◽  
Vitro Effect

Цель - исследование влияния глюкокортикоида дексаметазона (Dex) на процессы активации и генерации терминально-дифференцированных эффекторных CD4 Т-лимфоцитов (T) в культурах CD3CD45RO Т-клеток в условиях, имитирующих стимуляцию Т-клеточного рецептора in vitro в норме и при ревматоидном артрите (РА). Методика. Исследовали мононуклеарные лейкоциты (МНК) из венозной гепаринизированной крови 50 пациентов с ревматоидным артритом. Методом проточной цитофлюориметрии проанализировано изменение иммунофенотипа Т-лимфоцитов; иммуноферментным анализом оценена секреция CD3CD45RO Т-клетками цитокина IL-2; методом полимеразной цепной реакции определена экспрессия мРНК генов hTERT, U2af1l4 и Gfi1 в CD3CD45RO Т-клетках. Результаты. Полученные данные демонстрируют, что in vitro на фоне TCR-активации CD3CD45RO культур Т-клеток, Dex (в разной степени выраженности) участвует в формировании субпопуляции терминально-дифференцированных эффекторов (CD3CD4CD45ROCD28 T), характеризующихся низкой теломеразной активностью, потерей молекул костимуляции (CD28) и активации (CD25) и реэкспрессирующих высокомолекулярную изоформу рецептора CD45 - CD45RA в норме и у больных РА. Заключение. Сделано заключение, что популяция CD3CD4CD45ROCD28 лимфоцитов является ключевым участником в патогенезе РА, ускоренно приобретающая свой негативный потенциал на фоне глюкокортикоидной терапии, способствуя прогрессии заболевания, в частности, за счет мощного выброса провоспалительных медиаторов. Aim. To study the effect of dexamethasone (Dex) on activation and generation of terminally differentiated effector CD4 T-lymphocytes (TEMRA) in cultured CD3CD45RO T cells under the conditions mimicking stimulation of the T-cell receptor in vitro as it occurs in health and rheumatoid arthritis (RA). Methods. The study was performed on mononuclear leukocytes isolated from heparinized venous blood of 50 patients with rheumatoid arthritis. Changes in the T-lymphocyte immunophenotype were detected using flow cytofluorometry. Secretion of cytokine IL-2 by CD3 CD45ROT cells was assessed by ELISA. Expression of hTERT, U2af1l4, and Gfi1 gene mRNA in CD3CD45RO T cells was measured by polymerase chain reaction. Results. During the in vitro TCR activation of cultured CD3CD45RO T cells, Dex participated to a variable extent in formation of a subpopulation of terminally differentiated effectors (CD3CD4CD45ROCD28 TEMRA), which are characterized by low telomerase activity, loss of costimulation (CD28) and activation (CD25) molecules, and re-expression of the high molecular weight CD45-CD45RA receptor isoform both in healthy individuals and RA patients. Conclusion. The population of CD3CD4CD45ROCD28 lymphocytes is a key participant in the pathogenesis of RA by accelerating their negative impact during the glucocorticoid therapy. This lymphocyte population contributes to RA progression particularly due to the powerful discharge of proinflammatory mediators.

scholarly journals Program death-1 signaling and regulatory T cells collaborate to resist the function of adoptively transferred cytotoxic T lymphocytes in advanced acute myeloid leukemia

Blood ◽  
2010 ◽  
Vol 116 (14) ◽  
pp. 2484-2493 ◽  
Author(s):  
Qing Zhou ◽  
Meghan E. Munger ◽  
Steven L. Highfill ◽  
Jakub Tolar ◽  
Brenda J. Weigel ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Acute Myeloid Leukemia ◽  
Regulatory T Cells ◽  
Myeloid Leukemia ◽  
Antibody Treatment ◽  
T Effector Cells ◽  
Treg Depletion ◽  
Acute Myeloid

Abstract Tumor-induced immune defects can weaken host immune response and permit tumor cell growth. In a systemic model of murine acute myeloid leukemia (AML), tumor progression resulted in increased regulatory T cells (Treg) and elevation of program death-1 (PD-1) expression on CD8+ cytotoxic T cells (CTLs) at the tumor site. PD-1 knockout mice were more resistant to AML despite the presence of similar percentage of Tregs compared with wild type. In vitro, intact Treg suppression of CD8+ T-cell responses was dependent on PD-1 expression by T cells and Tregs and PD-L1 expression by antigen-presenting cells. In vivo, the function of adoptively transferred AML-reactive CTLs was reduced by AML-associated Tregs. Anti–PD-L1 monoclonal antibody treatment increased the proliferation and function of CTLs at tumor sites, reduced AML tumor burden, and resulted in long-term survivors. Treg depletion followed by PD-1/PD-L1 blockade showed superior efficacy for eradication of established AML. These data demonstrated that interaction between PD-1 and PD-L1 can facilitate Treg-induced suppression of T-effector cells and dampen the antitumor immune response. PD-1/PD-L1 blockade coupled with Treg depletion represents an important new approach that can be readily translated into the clinic to improve the therapeutic efficacy of adoptive AML-reactive CTLs in advanced AML disease.

Increase of CD4+CD25high Regulatory T Cells in the Peripheral Blood of Acute Myeloid Leukemia Patients.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 2546-2546
Author(s):  
Xingbing Wang ◽  
Junxia Yao ◽  
Jine Zheng ◽  
Jun Liu ◽  
Yanli He ◽  
...  
Keyword(s):  
Immune Response ◽  
T Cells ◽  
Acute Myeloid Leukemia ◽  
Regulatory T Cells ◽  
Peripheral Blood ◽  
Myeloid Leukemia ◽  
Normal Donor ◽  
Dependent Manner ◽  
Acute Myeloid

Abstract PURPOSE: Although immunosuppression has long been recognized in acute myeloid leukemia (AML), the underlying basis for the lack of an effective immune response against the tumor remains unclear. T cells constitutively expressing both CD4 and CD25 are essential for maintenance of self-tolerance and therefore have been referred to as regulatory T cells (Treg). Experimental tumor models in mice revealed that Tregs are potent inhibitors of an antitumor immune response. The purpose of the current study was to demonstrate the possible involvement of CD4+CD25high regulatory T cells in immune system impairment in patients with acute myeloid leukemia. METHODS: The frequency and phenotypes of CD4+CD25high T cells in the peripheral blood of 36 patients suffering from acute myeloid leukemia and from 15 healthy controls were determined by flow cytometry. To assess the functional activity of CD4+CD25high cells, CD4+CD25high or CD4+CD25− cells were purified from PBMCs by sorting with FACSVantage. The immunoregulatory properties of CD4+CD25high and CD4+CD25− T cells were characterized by proliferation and suppression assays. Also, cytokine production after in vitro stimulation was examined in CD4+CD25high and/or CD4+CD25− T cells isolated from AML patients. In addition, the frequency of apoptotic or proliferating T cells in CD4+CD25high T cells was determined by 7AAD or ki67 binding cells. RESULTS: Compared with healthy volunteers, patients with acute myeloid leukemia had a higher proportion of CD4+CD25high T cells (4.1±1.8% vs 2.0±0.5%) in peripheral blood with characteristics of Tregs, i.e., they are CD45-RA(−), CD69(−), CD45-RO(+), CD95(+), and intercellular CTLA-4(+), and secreted TGF-, TNF-α and IL-10 but did not secrete IL-2, IL-4, IL-5 or IFN-. When cocultured with CD4+25− cells, CD4+CD25high T cells potently suppressed their proliferation through a dose-dependent manner, thus behaving as Treg. In addition, In order to explain the increased prevalence of CD4+CD25high T cells in AML patients, we examined the apoptosis and proliferation variations of CD4+CD25high T cells in AML patients. Notably, in AML patients, both apoptosis and proliferation of CD4+CD25high T cells were significantly higher than that did normal donor. CONCLUSIONS: we provide evidence of an increased pool of CD4+CD25high regulatory T cells in the peripheral blood of AML patients with potent immunosuppressive features, which maybe due to the proliferation of CD4+CD25high T cells. This finding suggests that the use of immunomodulatory therapy to treat patients with AML may be an effective strategy.

Depletion of In-Vivo Activated Regulatory T Cells Followed by Synchronized Specific In-Vitro Activation Allows the Rapid Isolation of Leukemia-Reactive T Cells for Immunotherapy.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 297-297
Author(s):  
I. Jedema ◽  
E. Steeneveld ◽  
M. Hoogendoorn ◽  
R. Willemze ◽  
J.H.F. Falkenburg
Keyword(s):  
Immune Response ◽  
T Cells ◽  
T Cell ◽  
Regulatory T Cells ◽  
Cytotoxic Activity ◽  
High Frequency ◽  
Activated T Cells ◽  
Cell Clones

Abstract By CD40 crosslinking in the presence of cytokines leukemic cells can be modified into good antigen presenting cells (APC) expressing costimulatory molecules CD40, CD80, CD86, and CD83. Previously, primary alloreactive T cell responses from HLA-matched donors have been generated using these leukemic-APC as stimulator cells against acute and chronic myeloid leukemia (AML&CML), and acute and chronic lymphocytic leukemias (ALL&CLL). However, the likelihood of generating a good immune response is highly unpredictable and long-term culture in the presence of high dose IL-2 is needed to enrich for leukemia-reactive T cells. Since the length of the in-vitro culture period has been shown to be inversely correlated with the potential of cells to survive and expand in-vivo, we developed a method facilitating early activation, detection and rapid isolation of leukemia-reactive T cells based on their interferon-gamma (IFNg) secretion using the cytokine capture assay (Miltenyi). In order to enrich for leukemia-reactive T cells and to synchronize the production of IFNg, T cells were first stimulated with the leukemic APC with addition of low dose IL-2 (10 IU/mL) at day 7, resulting in re-entry of the majority of the T cells into a quiescent state after 14 days of culture. Then, the cells were specifically restimulated resulting in synchronized production of IFNg and allowing efficient isolation. Using this method we were able to isolate T cell populations containing a high frequency of leukemia-reactive T cells against CLL, ALL, AML, and CML in 11 donor/patient pairs. Using a CFSE-based cytotoxicity assay (Jedema, Blood2004; 103: 2677) as read-out we were able to demonstrate 20–80% lysis of the primary leukemic blasts by the IFNg+ T cells at very low E/T ratios (3/1-0.3/1) in the majority of the responses, whereas the IFNg- fractions induced only 5–30% lysis. Single cell sorting of the IFNg producing T cells revealed that 15–30% of the T cell clones was capable of exerting minor antigen specific cytotoxic activity against the patient cells in an HLA-restricted fashion. However, in individual cases despite minor antigen disparities between donor and patient no specific anti-leukemia immune response could be detected. Prior to exposure to the leukemic-APC in-vivo activated T cells were observed in the responder T cell population of these donors that contained a high frequency of regulatory T cells defined as CD4+/CD25+, CD4+/CD152+, and CD8+/CD28−. We hypothesized that these regulatory T cells might actively inhibit the induction of an anti-leukemic T cell response. Therefore, in a donor/CLL patient pair, in which we were not able to induce a cytotoxic immune response against the CLL-APC we removed the in-vivo activated T cells from the responder material prior to the initial activation with the CLL-APC. Whereas no cytotoxic activity could be isolated from unmodified responder material (only 1/288 clones was cytotoxic), the IFNg+ T cells isolated from the response induced after depletion of the in-vivo activated T cells was capable of exerting massive cytotoxicity against both the primary CLL (55%) and the CLL-APC (70%). Single cell cloning of this response revealed that 35/129 T cell clones (>25%, 26 CD8+, 9 CD4+) exerted HLA-restricted CLL-specific cytotoxicity. From these results we conclude that the likelihood of generating a primary anti-leukemic immune response is not only determined by the frequency of precursor CTLs, but also by the frequency of inhibitory regulatory T cells at the onset of the immune response.

scholarly journals MiR-146a regulates regulatory T cells to suppress heart transplant rejection in mice

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
Jian Lu ◽  
Weiwei Wang ◽  
Peiyuan Li ◽  
Xiaodong Wang ◽  
Chao Gao ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Heart Transplant ◽  
Transplant Rejection ◽  
Mouse Heart ◽  
Allograft Survival ◽  
Treg Function ◽  
Ifn Γ ◽  
Heart Transplant Rejection

AbstractRegulatory T cells (Tregs), which characteristically express forkhead box protein 3 (Foxp3), are essential for the induction of immune tolerance. Here, we investigated microRNA-146a (miR-146a), a miRNA that is widely expressed in Tregs and closely related to their homeostasis and function, with the aim of enhancing the function of Tregs by regulating miR-146a and then suppressing transplant rejection. The effect of the absence of miR-146a on Treg function in the presence or absence of rapamycin was detected in both a mouse heart transplantation model and cell co-cultures in vitro. The absence of miR-146a exerted a mild tissue-protective effect by transiently prolonging allograft survival and reducing the infiltration of CD4+ and CD8+ T cells into the allografts. Meanwhile, the absence of miR-146a increased Treg expansion but impaired the ability of Tregs to restrict T helper cell type 1 (Th1) responses. A miR-146a deficiency combined with interferon (IFN)-γ blockade repaired the impaired Treg function, further prolonged allograft survival, and alleviated rejection. Importantly, miR-146a regulated Tregs mainly through the IFN-γ/signal transducer and activator of transcription (STAT) 1 pathway, which is implicated in Treg function to inhibit Th1 responses. Our data suggest miR-146a controls a specific aspect of Treg function, and modulation of miR-146a may enhance Treg efficacy in alleviating heart transplant rejection in mice.

scholarly journals Trichostatin A Promotes the Generation and Suppressive Functions of Regulatory T Cells

2013 ◽  
Vol 2013 ◽  
pp. 1-8 ◽  
Author(s):  
Cristian Doñas ◽  
Macarena Fritz ◽  
Valeria Manríquez ◽  
Gabriela Tejón ◽  
María Rosa Bono ◽  
...  
Keyword(s):  
T Cells ◽  
Regulatory T Cells ◽  
Trichostatin A ◽  
Gene Promoter ◽  
Treg Cells ◽  
Specific Subset ◽  
Global Increase ◽  
And Function ◽  
H3 Acetylation

Regulatory T cells are a specific subset of lymphocytes that suppress immune responses and play a crucial role in the maintenance of self-tolerance. They can be generated in the thymus as well as in the periphery through differentiation of naïve CD4+T cells. The forkhead box P3 transcription factor (Foxp3) is a crucial molecule regulating the generation and function of Tregs. Here we show that thefoxp3gene promoter becomes hyperacetylated inin vitrodifferentiated Tregs compared to naïve CD4+T cells. We also show that the histone deacetylase inhibitor TSA stimulated thein vitrodifferentiation of naïve CD4+T cells into Tregs and that this induction was accompanied by a global increase in histone H3 acetylation. Importantly, we also demonstrated that Tregs generated in the presence of TSA have phenotypical and functional differences from the Tregs generated in the absence of TSA. Thus, TSA-generated Tregs showed increased suppressive activities, which could potentially be explained by a mechanism involving the ectonucleotidases CD39 and CD73. Our data show that TSA could potentially be used to enhance the differentiation and suppressive function of CD4+Foxp3+Treg cells.

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