Chronic Lymphocytic Leukaemia Drives the Production of T Regulatory Cells From the CD4+ CD25- Compartment with a CD27high/CD127 Low Phenotype.

Abstract Abstract 2330 Poster Board II-307 Patients with chronic lymphocytic leukaemia (CLL) have defects in both cellular and humoral immunity including changes in the numbers and function of T regulatory cells (Tregs). The identification of Tregs is an ever-evolving field and in this study we readdressed the phenotype using the markers CD25, FoxP3 and CD127-/lo and confirmed function by classical suppressor assays in CLL patients on and off treatment. Using the combination CD4 and FoxP3 we observed increased Treg frequencies in CLL patients, in particular with advanced disease, supporting previous studies showing an increase in Tregs in CLL. However in contrast to previous studies, there was no increase in the CD25+ FoxP3+ population in CLL patients rather the increase in FoxP3 expression occurred in the CD25- compartment of CLL patients. Interestingly CLL induced a 7-fold increase in the expression of FoxP3 in CD4+CD25- T cells following short-term co-culture. The T regulatory cells in CLL patients had a significantly higher expression of CD27 compared to healthy controls and although CD127 expression was low in both healthy and CLL patients it was significantly lower in CLL patients. Fludarabine treatment initially induced increased expression of FoxP3 in the CD4+ T cell compartment but this declined gradually to reach levels below that pre-treatment. Here we propose that CLL drives the production of Tregs from the CD4+CD25- compartment as has been shown recently in Non-Hodgkins Lymphoma (NHL) and the mechanism of induction could provide alternative avenues for treatment. Disclosures: No relevant conflicts of interest to declare.

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Deficient CD4+ CD25+ FOXP3+ T regulatory cells in acquired aplastic anemia

Blood ◽

10.1182/blood-2007-01-066258 ◽

2007 ◽

Vol 110 (5) ◽

pp. 1603-1606 ◽

Cited By ~ 126

Author(s):

Elena E. Solomou ◽

Katayoun Rezvani ◽

Stephan Mielke ◽

Daniela Malide ◽

Keyvan Keyvanfar ◽

...

Keyword(s):

T Cells ◽

Aplastic Anemia ◽

T Regulatory Cells ◽

T Cell Development ◽

Foxp3 Expression ◽

Mrna Levels ◽

Regulatory Cells ◽

Protein Levels ◽

Almost All ◽

And Function

Abstract Regulatory T cells are believed to control the development and progression of autoimmunity by suppressing autoreactive T cells. Decreased numbers of CD4+CD25+ FOXP3+ T cells (Tregs) are associated with impaired immune homeostasis and development of autoimmune diseases. The transcription factors FOXP3 and NFAT1 have key roles in regulatory T-cell development and function. We show that Tregs are decreased at presentation in almost all patients with aplastic anemia; FOXP3 protein and mRNA levels also are significantly lower in patients with aplastic anemia and NFAT1 protein levels are decreased or absent. Transfection of FOXP3-deficient CD4+CD25+ T cells from patients with a plasmid encoding wild-type NFAT1 resulted in increased FOXP3 expression in these cells. By NFAT1 knockdown in CD4+CD25+ T cells, FOXP3 expression was decreased when NFAT1 expression was decreased. Our findings indicate that decreased NFAT1 could explain low FOXP3 expression and diminished Treg frequency in aplastic anemia. Treg defects are now implicated in autoimmune marrow failure.

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N-Acetyl-β-D-Glucosaminidase Enzymuria in Leukaemia and Myelomatosis: Effect of Treatment in Acute Myeloblastic Leukaemia and Myelomatosis in Adults

Annals of Clinical Biochemistry International Journal of Laboratory Medicine ◽

10.1177/000456328602300609 ◽

1986 ◽

Vol 23 (6) ◽

pp. 676-680 ◽

Cited By ~ 1

Author(s):

P H Whiting ◽

D J King ◽

A Ireland ◽

M A Ratcliffe ◽

A A Dawson

Keyword(s):

Chronic Lymphocytic Leukaemia ◽

Lymphocytic Leukaemia ◽

Normal Values ◽

Terminal Phase ◽

Acute Myeloblastic Leukaemia ◽

Lysosomal Hydrolase ◽

Patient Groups ◽

Effect Of Treatment ◽

Pre Treatment ◽

Serum And Urine

The activity of the lysosomal hydrolase N-acetyl-β-d-glucosaminidase (NAG) was measured in the urine of patients with leukaemia or myeloma. Elevated pre-treatment enzymuria was noted in all patient groups with acute myeloblastic leukaemias (AML) FAB type M4 or 5 displaying higher activities than AML patients FAB types M1–3, which in turn were higher than those found in patients with myelomatosis and chronic lymphocytic leukaemia. The ratio of the major isoenzymes of NAG, A/B was reduced significantly only in patients with AML. Following treatment, AML patients who entered remission demonstrated NAG levels which approached normal values. In those AML patients who were either in relapse, in the terminal phase of their illness or treated with aminoglycoside antibiotics, NAG enzymuria was similar to pre-treatment values. A reduction in urinary NAG levels and both serum and urine β2 microglobulin concentrations was also observed following treatment in myeloma patients. The use of enzymuria both as a guide to progress towards remission in AML patients and for assessing prognosis and progress in myeloma patients is discussed.

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CD18 Is Required for Optimal Development and Function of CD4+CD25+ T Regulatory Cells

The Journal of Immunology ◽

10.4049/jimmunol.175.12.7889 ◽

2005 ◽

Vol 175 (12) ◽

pp. 7889-7897 ◽

Cited By ~ 64

Author(s):

Marissa Marski ◽

Sravanthi Kandula ◽

Jerrold R. Turner ◽

Clara Abraham

Keyword(s):

T Regulatory Cells ◽

Regulatory Cells ◽

Optimal Development ◽

And Function

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Dexamethasone affects day/night development and function of thymus-derived T regulatory cells

Immunobiology ◽

10.1016/j.imbio.2019.07.007 ◽

2019 ◽

Vol 224 (5) ◽

pp. 614-624

Author(s):

Ewelina Kiernozek ◽

Anna Bieńkowska ◽

Magdalena Markowska ◽

Ewa Kozlowska ◽

Nadzieja Drela

Keyword(s):

T Regulatory Cells ◽

Regulatory Cells ◽

And Function

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Pertussis toxin as an adjuvant suppresses the number and function of CD4+CD25+ T regulatory cells

European Journal of Immunology ◽

10.1002/eji.200535353 ◽

2006 ◽

Vol 36 (3) ◽

pp. 671-680 ◽

Cited By ~ 77

Author(s):

Xin Chen ◽

Robin T. Winkler-Pickett ◽

Nicholas H. Carbonetti ◽

John R. Ortaldo ◽

Joost J. Oppenheim ◽

...

Keyword(s):

Pertussis Toxin ◽

T Regulatory Cells ◽

Regulatory Cells ◽

And Function

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Runx1 and Runx3 Are Involved in the Generation and Function of Highly Suppressive IL-17-Producing T Regulatory Cells

PLoS ONE ◽

10.1371/journal.pone.0045115 ◽

2012 ◽

Vol 7 (9) ◽

pp. e45115 ◽

Cited By ~ 19

Author(s):

Lequn Li ◽

Nikolaos Patsoukis ◽

Victoria Petkova ◽

Vassiliki A. Boussiotis

Keyword(s):

T Regulatory Cells ◽

Regulatory Cells ◽

And Function

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The anti-cancer agents lenalidomide and pomalidomide inhibit the proliferation and function of T regulatory cells

Cancer Immunology Immunotherapy ◽

10.1007/s00262-008-0620-4 ◽

2008 ◽

Vol 58 (7) ◽

pp. 1033-1045 ◽

Cited By ~ 254

Author(s):

Christine Galustian ◽

Brendan Meyer ◽

Marie-Christine Labarthe ◽

Keith Dredge ◽

Deborah Klaschka ◽

...

Keyword(s):

T Regulatory Cells ◽

Regulatory Cells ◽

Anti Cancer ◽

And Function

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To Treat or Not to Treat: Metabolomics Reveals Biomarkers for Treatment Indication in Chronic Lymphocytic Leukaemia Patients

Blood ◽

10.1182/blood.v126.23.5286.5286 ◽

2015 ◽

Vol 126 (23) ◽

pp. 5286-5286

Author(s):

Jaroslaw Piszcz ◽

Emily Grace Armitage ◽

Ferrarini Alessia ◽

Francisco Javier Ruperez ◽

Agnieszka Kulczynska ◽

...

Keyword(s):

Chronic Lymphocytic Leukaemia ◽

Lymphocytic Leukaemia ◽

Conflicts Of Interest ◽

Disease Onset ◽

Disease Status ◽

Analytical Technique ◽

Fatty Acid Amides ◽

Staging Systems ◽

Disease Staging ◽

Patient Survival Time

Abstract In chronic lymphocytic leukaemia (CLL) clinical course of patients is heterogeneous. Some present an aggressive disease onset and require immediate therapy, while others remain without treatment for years. Current disease staging systems developed by Rai and Binet may be useful in forecasting patient survival time, but do not discriminate between stable and progressive forms of the disease in the early stages. Recently ample attention has been directed towards identifying new disease prognostic markers capable of predicting clinical aggressiveness at diagnosis. In this research we reached for LC-MS metabolic fingerprinting method to analyse serum of stable (n=51) and progressive (n=42) CLL patients and controls (n=45) with aim to discover metabolic indicators of disease status. A panel of markers discriminating aggressive from indolent patients was discovered. Ten of them were selected for validation on larger population (45 controls and 92 CLL) with an independent analytical technique. Linoleamide (p=0.002) in addition to various acylcarnitines (p=0.001-0.000001) showed to be significant markers of CLL in its aggressive form. Acetylcarnitine (p=0.05) and hexannoylcarnitine (p=0.005) were also distinguishable markers of indolent subjects. Forming a panel of selected acylcarnitines and fatty acid amides, it was possible to reach a highly specific and sensitive diagnostic approach (AUC=0.766). Disclosures No relevant conflicts of interest to declare.

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Granzyme B Expression in T-Regulatory Cells Is a Strong Predictor of Acute Graft-Versus-Host Disease after Day +30 in Patients with "Classic" Immunosuppression after Allo-HSCT

Blood ◽

10.1182/blood.v128.22.2238.2238 ◽

2016 ◽

Vol 128 (22) ◽

pp. 2238-2238

Author(s):

Mikhail Yu. Drokov ◽

Elena N. Parovichnikova ◽

Julia Davydova ◽

Larisa A. Kuzmina ◽

Irina V Galtseva ◽

...

Keyword(s):

Roc Curve ◽

T Regulatory Cells ◽

Conflicts Of Interest ◽

Granzyme B ◽

Conditioning Regimen ◽

Curve Analysis ◽

Unrelated Donor ◽

Regulatory Cells ◽

Roc Curve Analysis ◽

Becton Dickinson

Abstract Introduction. Granzyme B is a serine protease commonly found in the granules of cytotoxic lymphocytes and natural killer cells. It is secreted with the pore forming protein perforin and mediates apoptosis in target cells. Granzyme B mediated cytolysis is one of the regulatory mechanisms (together with IL-2 receptors (CD25)) by which T-regulatory cells (T-regs) influence on T-effectors cells. Despite the well-known fact that T-regs participate in pathogenesis of aGVHD and their amount after allo-HSCT inversely correlates with the probability of aGVHD incidence, data about functional status of T-regs and aGVHD is still limited. Patients and methods. Peripheral blood samples were collected in EDTA-tubes at day +30 after allo-HSCT. We use PBMC from 29 patients with hematological malignancies obtained by density gradient media. This method was used due to lymphopenia in this group of patients. Group with no aGVHD consist of 22 patients (AML=12, ALL n=5, LPD=1, CML=2, AA=1, MDS=1) after allo-HSCT n=17 from MUD, n=5 from mismatch unrelated donor. MAC conditioning regimen was used in 17 cases, 5 patients receive RIC. The group of aGVHD after day +30 include 7 patients. (AML=4, ALL n=3) after allo-HSCT from MUD (n=5), mismatch unrelated donor (n=1) and n=1 from sibling HLA-identical donor. Two patients receive MAC and 5 patients receive RIC conditioning regimen. All patients in both groups receive standard immunosuppression (MMF+CSA+ATG). All patients developed II-IV grade aGVHD (II (n=1); III (n=4); IV (n=2)) with a median time onset on day +50 (34-150). The anti-CD4-APC-Cy7, anti-CD25-APC, anti-CD127-FITC and anti-Granzyme B-PE (Becton Dickinson, USA) antibodies were used to determine T-regulatory cells population. A Bland-Altman plot (difference plot) was used in analyzing the agreement between the two different methods of T-regs assay (CD4+CD25high and CD4+CD25highCD127low) - differences were not found (p=0,942). Due to this fact in our experiments CD4+CD25high cells were identified as T-reg cells (Gregg et al., 2005). 30000 of CD4+ cells were analyzed on a BD FACSCanto II to achieve sufficient statistical power (Becton Dickinson, USA). Results. As we can see on chart 1 level of Granzyme B was higher in patients who never developed aGVHD. In accordance with chart 2 percentage of Granzyme B positive T-regs in group of patients who never developed aGVHD was 7,26±1,89% in comparison with 2,04±0,93% in group who developed aGVHD after day +30 (p=0,02*). We should note that according to our previous experiments bacterial or viral infection (e.g CMV) and HLA-disparity does not affect the level of granzyme B expression in T-regs. Using ROC- curve analysis (see Chart 3) we obtained area under curve (AUC) 0,74. Conclusion. Our data shows that Granzyme B in T-reg cells after allo-HSCT in patients with standard IST may predict aGVHD onset after day +30 with satisfactory test results (AUC=0,74, «cut-off» - 4,15%; sensitivity - 85,71%; specificity - 45,45%).). According to data of our transplant center (for 2006-2016), "granzyme B-based strategy" can help us to predict up to 47,3% of all aGVHD. Disclosures: No relevant conflicts of interest to declare. Chart 1: Example of Granzyme B expression (right; shown on horizontal axis) in T-regs (left; upper right quadrant) on day +30 after allo-HSCT in group with standard immunosuppression Chart 2: Percentage of Granzyme B positive T-reg cells in patient without aGVHD and patients who develop aGVHD after day +30. Chart 3: ROC curve analysis of Granzyme B in T-regs as predictor of aGVHD. Figure Figure. Figure Figure. Figure Figure. Disclosures No relevant conflicts of interest to declare.

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