Effects Of Low-dose Interleukin-2 Research Articles

The effect of low-dose interleukin-2 on the T cell receptor landscape in patients with acute myocardial infarction

The effect of low-dose interleukin-2 on the T cell receptor landscape in patients with acute myocardial infarction

Low Dose IL-2 Therapy Induces T Regulatory Cell Derived Circulatory Exosomes Containing PDL1 and CD73 and Abrogates Development of Chronic Cardiac Allograft Rejection

<h3>Purpose</h3> Aim of the study is to determine the effect of low dose interleukin-2 (IL-2) therapy on the kinetics of induction of circulating exosomes and their immunological properties resulting in prevention of chronic rejection following allogeneic murine cardiac transplantation (HTx). <h3>Methods</h3> BALB/c (H2^d) to C57BL/6 (H2^b) HTx was performed heterotopically in the abdominal cavity. Costimulatory blockade consisting of MR1 (250 μg/ip day 0) and CTLA4-Ig (200 μg/ip day 2) was administered to induce tolerance. IL-2 (2000IU/day) was administered from day 14 post-transplant for 3 weeks. Sera were collected on days 7-100 for analysis of exosome induction and characterization of cardiac self-antigens (SAgs) (Myosin, Vimentin) by western blot. Foxp3, PDL1, CD73 in exosomes were detected using aldehyde bead - based flow cytometry. Antibodies (Abs) to donor MHC and SAgs were detected using immunofluorescence and ELISA respectively. <h3>Results</h3> BALB/c to C57BL/6 HTx acutely rejected (AR) with median survival time of 8 days. Costimulatory blockade prevented AR but developed chronic rejection with mean of 42 days circulating exosomes contained significantly higher levels of Myosin and Vimentin in the exosomes day 15 (p<0.05) and day 30 (p<0.01). Abs to MHC (32±13.8% vs 2±1.1; p<0.001) was detected on day 30, whereas Abs to Myosin (1114.21±443.4 ng/ml; p<0.0001) and Vimentin (519±241 ng/ml; p<0.0001) were detected on day45 during chronic rejection with fibrosis (57.1±20.7 vs 2.3±1.7%). Low dose IL-2 significantly prolonged the survival, 115 vs 42 (p<0.001) and did not induce exosomes with cardiac SAgs or developed Abs to MHC or SAgs. Low dose IL-2 not only abrogated chronic rejection but also induced splenic CD4+CD25+ Foxp3 T regulatory (Treg) cells in the allograft by day 45. Flow cytometry of circulatory exosomes isolated in day 45 also demonstrated increased Foxp3+ PDL1+ (3.3 fold) and FoxP3+ CD73+ (3 fold) compared to exosomes isolated on day 45 from chronic rejecting animals. <h3>Conclusion</h3> Our results demonstrate that low dose IL-2 abrogated the development of chronic rejection following HTx and induced circulating Treg cells derived exosomes with PDL1 and CD73 along with splenic Tregs and graft infiltrating FoxP3 cells.

Low Dose Interleukin-2 Induces Exosomes with Tolerance Markers (PDL1, CD73) and Significantly Delays Development of Chronic Rejection Following Murine Heterotopic Cardiac Transplantation

Purpose Aim of the study is to determine the effect of low dose interleukin-2 (IL-2) therapy on the kinetics of induction of circulating exosomes with tolerance markers (PDL1, CD73) and their role in abrogating chronic rejection following murine heterotopic cardiac transplant (HTx). Methods Allogenic (BALB/c to C57BL/6) HTx was performed heterotopically in abdominal cavity. Costimulatory blockade consisting of MR1 (250 μg/ip day 0) and CTLA4-Ig (200 μg/ip day 2) was performed. Low dose IL-2 (2000IU/day) was administered from day 15 post-transplant for 3 weeks. Sera was collected on days 7-100 for kinetic analysis of exosome induction and characterization of PDL1, CD73, cardiac self-antigens (SAgs) (Myosin, Vimentin), class II MHC trans-activator (CIITA), and 20S proteasome by western blot using specific antibodies (Abs) and Abs to SAgs. Results BALB/c to C57BL/6 HTx rejected transplanted heart with median survival time of 8 (7-9) days. HTx done following costimulatory blockade developed chronic rejection with mean survival time 26 (7-47) days. Exosome and Ab development demonstrated induction of exosomes with Vimentin (2.2± 1.0 vs 0.6±0.1; p Conclusion Circulating exosomes containing cardiac SAgs and development of Abs to SAgs lead to chronic rejection in a murine model occurring after costimulation blockade following allogenic HTx. Low dose IL-2 abrogated chronic rejection and induced circulating exosome with tolerance markers. This result supports an important role for exosomes, inducing tolerance and significantly delaying development of chronic rejection in murine model of chronic cardiac allograft rejection.

SAT0278 LOW-DOSE IL-2 RESTORES TREG-MEDIATED IMMUNE TOLERANCE IN PATIENTS WITH ANCA-ASSOCIATED VASCULITIS

Background:Antineutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV) is a group of autoimmune disease that can cause systemic organ damage, including granulomatosis with polyangiitis(GPA), microscopic polyangiitis (MPA), and eosinophilic granulomatosis with polyangiitis(EGPA)[1]. Several observations have showed that the breakdown of immune tolerance was involved in the pathogenesis of AAV [2], furthermore, a single, open and clinical trial demonstrates that IL-2 can be used to treat patients with GPA [3]. But there is still a lack of understanding of the relationship between Th17 / Treg and AAV and evidence for the therapeutic effect of IL-2 on AAV, which needs further exploration.Objectives:We first measured the absolute number of CD4+T subsets in peripheral blood of patients to explore the pathogenesis of AAV, and then investigated the effects of short-term and low-dose recombinant human IL-2 (rhIL-2) on CD4+T subsets of patients to analyze the regulatory effect of IL-2 on AAV.Methods:49 patients with AAV, hospitalized at the Second Hospital of Shanxi Medical University from the May 2016 to the November 2019 were enrolled, including 36 patients who were only received conventional glucocorticoids and DMARDs, and other 13 patients who were not only received these treatments but were also injected subcutaneously rhIL-2(50WIU/day for a 5-day course). 31 age and gender-matched healthy adults were selected as controls. The absolute number of Th17 and Treg cells in peripheral blood of health controls and the patients before and after treatment was detected by flow cytometry.Results:There was significant decreased level of Treg cells in the patients with AAV compared with healthy controls (P&lt;0.001) leading to a higher Th17/Treg ratio in the patients with AAV, but there was no statistically significant in the absolute number of Th17 cells between the patients and healthy controls. After the treatment of short-term and low-dose IL-2, there was a significant increase in the absolute number of Treg cells (P&lt;0.01) leading to a decrease in the ratio of Th17 and Treg (p&lt;0.05).The absolute number of Th17 had a trend towards higher values but was not statistical significance.Conclusion:The difference of Treg cells between the patients and healthy controls suggested that the decreased number of Treg cells failed to control autoimmune inflammatory response contributing to the pathogenesis of AAV. After the treatment of short-term and low-dose rhIL-2, there was a more significant increase in the absolute number of Treg cells showing that IL-2 could selectively stimulate the growth of Treg cells and restore the Treg-mediated immune tolerance in patients with AAV to achieve disease remission.

Interleukin-2 Therapy of Autoimmunity in Diabetes (ITAD): a phase 2, multicentre, double-blind, randomized, placebo-controlled trial

Type 1 diabetes is a common autoimmune disease due to destruction of pancreatic β cells, resulting in lifelong need for insulin. Evidence suggest that maintaining residual β-cell function can improve glucose control and reduce risk of hypoglycaemia and vascular complications. Non-clinical, preclinical and some preliminary clinical data suggest that low-dose interleukin-2 (IL-2) therapy could block pancreatic β cells destruction by increasing the number of functional regulatory T cells (Tregs) that inhibit islet-specific autoreactive effector T cells (Teffs). However, there is lack of data on the effect of low-dose IL-2 in newly diagnosed children and adolescents with T1D as well as lack of specific data on its potential effect on β-cell function. The ‘ Interleukin-2 Therapy of Autoimmunity in Diabetes (ITAD)’ is a phase 2, multicentre, double-blind, randomised, placebo-controlled trial in children and adolescents (6-18 years; having detectable C-peptide) initiated within 6 weeks of T1D diagnosis. A total of 45 participants will be randomised in a 2:1 ratio to receive either ultra-low dose IL-2 (aldesleukin), at a dose of 0.2 x 10 6 IU/m 2 twice-weekly, given subcutaneously, or placebo, for 6 months. The primary objective is to assess the effects of ultra-low dose aldesleukin administration on endogenous β-cell function as measured by frequent home dried blood spot (DBS) fasting and post-prandial C-peptide in children and adolescents with newly diagnosed T1D. The secondary objectives are: 1) to assess the efficacy of regular dosing of aldesleukin in increasing Treg levels; 2) to confirm the clinical safety and tolerability of ultra-low dose aldesleukin; 3) to assess changes in the immune system indicating benefit or potential risk for future gains/loss in β-cell function and immune function; 4) to assess treatment effect on glycaemic control. Trial registration: EudraCT 2017-002126-20 (06/02/2019)

Interleukin-2 Therapy of Autoimmunity in Diabetes (ITAD): a phase 2, multicentre, double-blind, randomized, placebo-controlled trial.

Type 1 diabetes is a common autoimmune disease due to destruction of pancreatic β cells, resulting in lifelong need for insulin. Evidence suggest that maintaining residual β-cell function can improve glucose control and reduce risk of hypoglycaemia and vascular complications. Non-clinical, preclinical and some preliminary clinical data suggest that low-dose interleukin-2 (IL-2) therapy could block pancreatic β cells destruction by increasing the number of functional regulatory T cells (Tregs) that inhibit islet-specific autoreactive effector T cells (Teffs). However, there is lack of data on the effect of low-dose IL-2 in newly diagnosed children and adolescents with T1D as well as lack of specific data on its potential effect on β-cell function. The ' Interleukin-2 Therapy of Autoimmunity in Diabetes (ITAD)' is a phase 2, multicentre, double-blind, randomised, placebo-controlled trial in children and adolescents (6-18 years; having detectable C-peptide) initiated within 6 weeks of T1D diagnosis. A total of 45 participants will be randomised in a 2:1 ratio to receive either ultra-low dose IL-2 (aldesleukin), at a dose of 0.2 x 10 6 IU/m 2 twice-weekly, given subcutaneously, or placebo, for 6 months. The primary objective is to assess the effects of ultra-low dose aldesleukin administration on endogenous β-cell function as measured by frequent home dried blood spot (DBS) fasting and post-prandial C-peptide in children and adolescents with newly diagnosed T1D. The secondary objectives are: 1) to assess the efficacy of regular dosing of aldesleukin in increasing Treg levels; 2) to confirm the clinical safety and tolerability of ultra-low dose aldesleukin; 3) to assess changes in the immune system indicating benefit or potential risk for future gains/loss in β-cell function and immune function; 4) to assess treatment effect on glycaemic control. Trial registration: EudraCT 2017-002126-20 (06/02/2019).

Effect of low dose IL-2 loaded chitosan nanoparticles on natural killer and regulatory T cell expression in experimentally induced autoimmune type 1 diabetes mellitus.

IntroductionNatural killer cells (NK) initiate pancreatic islets cell lyses in autoimmune type 1 diabetes mellitus (T1D). Loss of T regulatory cells (Treg) at disease onset facilitates activation and accumulation of NKs in the pancreatic microenvironment. A proper low dose interleukin 2 (IL-2) could enhance Tregs and enforce control and regulation of pro-inflammatory NKs. This relation needs to be studied to improve therapeutic strategies aimed at resetting the balance between Tregs and proinflammatory cells.Material and methodsWe used novel formulations of low dose IL-2 loaded on chitosan nanoparticles. The study included 116 T1D BALB/c mice experimentally induced by streptozotocin, divided into groups. Their splenocytes were maintained in a short-term culture for assessment of expression of CD4+Foxp3+ Treg and NKp46+NK by both flow cytometry and enzyme linked immunoassay (ELISA). In vitro suppressor-assay was used in order to assess the suppressor effect of Treg cells after exogenous IL-2 treatment.ResultsNK cell expression, NKp46 level and NK cell functions were modulated in mice injected with IL-2 loaded chitosan nanoparticles than other groups. A statistical inverse correlation was found between Treg and NK cell expression in IL-2 loaded chitosan with (0.3 µIU) (p = 0.047) and this correlation was related to Foxp3 expression on Treg cells. The modified expression of NK and NKp46 was noticed in mice injected with (0.3 µIU) for longer duration (three weeks) (p < 0.001) but the NK functions did not show any significant changes with prolonged treatment.ConclusionsLow dose (0.3) µIU IL-2 nanoparticles effectively modulated NK and NKp46 expression. It selectively modulates the suppressive activity of Tregs indicating a significant role of Tregs in NK activation and function by controlling the availability of IL-2 in the microenvironment.

The absolute counts of peripheral T lymphocyte subsets in patient with ankylosing spondylitis and the effect of low-dose interleukin-2.

The aim of this analysis is to investigate the level of T cell subsets in ankylosing spondylitis (AS) and the effect of low-dose interleukin 2 (IL-2).This is a retrospective cohort study, we collected basic information, inflammatory markers, BASDAI scores, and T lymphocyte subsets of peripheral blood in patients with AS, baseline analysis, correlation analysis and comparative analysis before and after treatment were performed.Data from 73 patients (of these, 36 patients were treated with IL-2) and 85 the health were included. The absolute numbers of peripheral CD4+ Treg and CD8+ Treg cells were lower, while the Th17 cell number and the ratio of Th17/CD4+ Treg and CD4+/CD8+ Treg were higher. The CD4+ Treg levels and the ratio of Th17/CD4+ Tregs were correlated with BASDAI. The CD4+ Treg, CD8+ Treg, and Th17 cells were increased after treatment with IL-2 and the ratio of Th17/CD4+ Treg was decreased.Increase in ratio of Th17/CD4+ Treg involved in the occurrence of AS. At the same time, low-dose IL-2 could decrease the ratio of Th17/CD4+ Treg in short time, low-dose IL-2 can be used to treat autoimmune diseases to avoid adverse reactions caused by immunosuppressants. Further clinical study is needed on the efficacy of IL-2.

Immunological and clinical effects of low-dose interleukin-2 across 11 autoimmune diseases in a single, open clinical trial

背景 调节性T（Tregs）细胞可以预防自身免疫并控制炎症，在各种自身免疫性或自身炎症性疾病中的研究都表明Treg细胞不足。由于低剂量白细胞介素-2（low-dose interleukin-2，ld-IL2）能够扩增并激活Tregs细胞，因此ld-IL2具有广泛的治疗潜力。 目的 我们通过一项临床试验观察ld-IL2治疗11种自身免疫性疾病中1种的效果，旨在评估ld-IL2的治疗潜力并选择疾病，以用于进一步的临床开发。 方法 本研究为前瞻开放性Ⅰ~Ⅱa期临床观察，入选了46例轻中度患者，诊断分别为类风湿关节炎、强直性脊柱炎、系统性红斑狼疮、牛皮癣、白塞病、肉芽肿性多血管炎、大动脉炎、克罗恩病、溃疡性结肠炎、自身免疫性肝炎或硬化性胆管炎。所有患者都接受了ld-IL2（100万IU/天）5天，然后每两周注射一次，维持6个月。所有患者均接受了深入的免疫监测和临床评估。 结果 在任何疾病类型以及任何合并药物治疗的患者中，ld-IL2都显示出了良好的耐受性。通过经典的免疫表型分析和广泛的免疫表型分析发现所有患者的Tregs细胞均被特异性地扩增激活，而效应T细胞没有被活化，提示ld-IL2具有临床疗效的前景。 结论 本研究中所采用的IL-2剂量和治疗方案可以选择性地激活和扩增Tregs细胞，并且对于不同的疾病种类以及不同的合并用药都是安全的。这一临床疗效以及初步结果可以为ld-IL2在各种自身免疫和自身炎症性疾病中开展Ⅱ期临床试验观察疗效提供了依据。 试验注册号 NCT01988506

A phase II trial of low-dose interleukin-2 (IL-2) and bevacizumab in patients with metastatic renal cell carcinoma (mRCC)

5103 Background: Interleukin-2 and bevacizumab, a VEGF ligand-binding antibody, have each demonstrated antitumor activity in mRCC. Increased VEGF levels can lead to immunosuppression and resistance to immunotherapy through inhibition of dendritic cell (DC) differentiation and increase in immunosuppressive regulatory T cells (Tregs). To further evaluate the clinical and immunomodulatory effect of low-dose IL-2 and bevacizumab in mRCC, a phase II trial was conducted. Methods: Previously untreated good and intermediate risk mRCC pts received 8-week cycles of IL-2 (250,000 U/kg/d SC D1–5 during week 1 and 125,000 U/kg/d SC D1–5 during weeks 2–6, followed by a 2 week break). Bevacizumab 10mg/kg was administered IV every 2 weeks starting on day -14. Eligibility included RECIST-defined measurable disease, clear cell histology, normal organ function, and prior nephrectomy. A Simon 2 stage phase II design was employed to test the hypothesis of a 40% improvement in the 3-month PFS vs. historical IL-2-treated controls. Overall response rate (ORR) and toxicity were recorded. Exploratory endpoints included activation of circulating DC’s, Tregs and VEGF levels. Results: To date 16 of a planned 35 pts are enrolled. Median age is 59 years (range, 44–67); Eight patients have &gt;1 site of metastasis; median treatment duration is 8 weeks (8–32+). Among 11 pts evaluable for response, 1 PR and 3 SD lasting &gt;3 months have been observed. All pts with SD have demonstrated some degree of tumor shrinkage. Seven pts have discontinued therapy (5 PD, 2 withdrew consent). Most common treatment-related toxicity included fatigue, nausea, diarrhea, and fever. When comparing immune correlates at day 56 of therapy vs. baseline, all pts had an increase in the number of Tregs (median increase 1.65, range; 0.76–10.4; p= 0.008). Similarly, all pts had a decline in VEGF levels (median decline 164.38 pg/ml, range; 2.20–393.04 pg/ml; p= 0.008). No differences in DC activation have been observed. Conclusions: The combination of LD IL-2 and bevacizumab produces antitumor activity and moderate toxicity. Preliminary correlative data demonstrates inhibition of VEGF and increase in Tregs without effect on DC activation. No significant financial relationships to disclose.

Effect of low-dose interleukin-2 and interferon alfa on t-lymphocyte subsets in melanoma

Effect of low-dose interleukin-2 and interferon alfa on t-lymphocyte subsets in melanoma

Effect of low-dose interleukin-2 on disease relapse after T-cell- depleted allogeneic bone marrow transplantation

T-cell depletion of donor bone marrow has been associated with an increased risk of disease relapse after allogeneic bone marrow transplantation (BMT). Recombinant interleukin-2 (IL-2), which is capable of increasing the antileukemic activity of peripheral blood lymphocytes obtained from patients who have undergone BMT, has been proposed as a potentially useful agent to reduce the risk of relapse post-BMT. We have previously shown that IL-2 administered to patients at very low doses after BMT is both clinically tolerable and immunologically active. We now report on the clinical outcome of 29 patients treated with low-dose IL-2 after CD6-depleted allogenic BMT for hematologic malignancy. IL-2 was administered by continuous infusion for up to 3 months beginning at a median of 67 days post-BMT. Eligibility requirements for IL-2 therapy included demonstration of stable engraftment and absence of acute grade 2–4 graft-versus-host disease (GVHD). Low-dose IL-2 was well tolerated by the majority of patients, with only 4 of 29 subjects withdrawn early. Acute GVHD developed in only one individual. After 12 weeks of treatment, the mean number of circulating natural killer cells in patients increased 10- fold without any significant change in T-cell number. Of the 25 patients who received &gt; or = 1 month of IL-2, only 6 have relapsed. Relapse rate and disease-free survival (DFS) were determined in the 25 patients who completed at least 4 weeks of IL-2 treatment and compared with historical controls transplanted at our institution for the same conditions and treated with an identical ablative regimen and method of T-cell depletion. Only control patients who had survived disease free for 100 days post-BMT were included in this analysis. Cox's proportional hazards regression model suggested that, compared with control patients without a history of GVHD, patients treated with IL-2 had a lower risk of disease relapse (hazard ratio 0.34; range, 0.14 to 0.82) and superior DFS (hazard ratio 0.39; range 0.18 to 0.87). A randomized controlled trial of IL-2 immunotherapy after T-cell-depleted BMT should now be undertaken.

Effect of Low-Dose Interleukin-2 on Disease Relapse After T-Cell-Depleted Allogeneic Bone Marrow Transplantation

T-cell depletion of donor bone marrow has been associated with an increased risk of disease relapse after allogeneic bone marrow transplantation (BMT). Recombinant interleukin-2 (IL-2), which is capable of increasing the antileukemic activity of peripheral blood lymphocytes obtained from patients who have undergone BMT, has been proposed as a potentially useful agent to reduce the risk of relapse post-BMT. We have previously shown that IL-2 administered to patients at very low doses after BMT is both clinically tolerable and immunologically active. We now report on the clinical outcome of 29 patients treated with low-dose IL-2 after CD6-depleted allogenic BMT for hematologic malignancy. IL-2 was administered by continuous infusion for up to 3 months beginning at a median of 67 days post-BMT. Eligibility requirements for IL-2 therapy included demonstration of stable engraftment and absence of acute grade 2-4 graft-versus-host disease (GVHD). Low-dose IL-2 was well tolerated by the majority of patients, with only 4 of 29 subjects withdrawn early. Acute GVHD developed in only one individual. After 12 weeks of treatment, the mean number of circulating natural killer cells in patients increased 10-fold without any significant change in T-cell number. Of the 25 patients who received > or = 1 month of IL-2, only 6 have relapsed. Relapse rate and disease-free survival (DFS) were determined in the 25 patients who completed at least 4 weeks of IL-2 treatment and compared with historical controls transplanted at our institution for the same conditions and treated with an identical ablative regimen and method of T-cell depletion. Only control patients who had survived disease free for 100 days post-BMT were included in this analysis. Cox's proportional hazards regression model suggested that, compared with control patients without a history of GVHD, patients treated with IL-2 had a lower risk of disease relapse (hazard ratio 0.34; range, 0.14 to 0.82) and superior DFS (hazard ratio 0.39; range 0.18 to 0.87). A randomized controlled trial of IL-2 immunotherapy after T-cell-depleted BMT should now be undertaken.

Recruitment of inflammatory cells to a tumor deposit potentiates the immunotherapeutic effects of interleukin-2.

Interleukin-2 (IL-2) is a potent immunotherapeutic agent in murine models of intraperitoneal, pulmonary, and hepatic tumor implantation. Because of the systemic toxicity documented at doses of IL-2 required to control tumor growth, potentiation of the effects of low dose IL-2 is an important problem in immunotherapy. To address this problem, we attempted to recruit lymphocytes into a tumor mass. Allogeneic P185 (H-2d) tumor was mixed with MCA-105 (H-2b) tumor and injected s. c. into C57BL/6 (H-2b) mice. Mice were treated with 50,000 units of IL-2 twice daily from day 0 to day 6. When IL-2 alone was used to treat s. c. tumor, there was no reduction in the size of tumor implants. When allogeneic tumor was mixed with syngeneic tumor, there was a reduction in tumor size at the high dose of allogeneic tumor but not at the low dose. When allogeneic tumor was mixed with syngeneic tumor and the mice treated with IL-2, the immunotherapeutic effects of IL-2 were markedly increased. These studies show that an immune response to alloantigens, generated within tumor tissue can augment the immunotherapeutic effects of exogenous IL-2.