Maria Skrzypkowska

Loss of the balance between CD4+Foxp3+ regulatory T cells and CD4+IL17A+ Th17 cells in patients with type 1 diabetes

The presence of low-grade chronic inflammation is a known feature of long standing diabetes type 1. Recently, two T cell subsets, that may contribute to the disease progression are under investigation. These are Treg cells, which are specialized T cell subset, that controls the activity of autoreactive and inflammatory cells and Th17 cells which are involved in the pathogenesis of inflammatory and autoimmune diseases. The balance between Treg and Th17 controls inflammation and is responsible for the proper function of the immune system. An decrease of Tregs and/or increase of Th17 may induce local inflammation, which in turn may hasten the development of diabetic complications. In the present study, we have demonstrated that the Treg/Th17 balance was broken in patients with diabetes type 1 and might contribute to the progression of microvascular angiopathy.

The Serum IL-6 Profile and Treg/Th17 Peripheral Cell Populations in Patients with Type 1 Diabetes

IL-6 is a pleiotropic cytokine involved in the regulation of the immune response, inflammation, and hematopoeisis. Its elevated levels are found in a range of autoimmune and chronic inflammatory diseases. IL-6 is also involved in regulation of the balance between two T cell subsets: Tregs and Th17, which have contradictory functions in the control of inflammation. The present study provides a quantitative analysis regarding the Th17/Treg cell balance in peripheral blood of children with type 1 diabetes and its association with serum IL-6 level.

IL-33 Effect on Quantitative Changes of CD4+CD25highFOXP3+ Regulatory T Cells in Children with Type 1 Diabetes.

IL-33 is an IL-1 cytokine family member, with ability to induce both Th1 and Th2 immune responses. It binds to ST2 receptor, whose deficiency is associated with enhanced inflammatory response. The most recent studies have shown the immunoregulatory effect of IL-33 on Tregs in animal models. As type 1 diabetes is an autoimmune, inflammatory disease, where Treg defects have been described, we aimed to analyze the in vitro influence of recombinant IL-33 on quantitative properties of regulatory CD4+CD25highFOXP3+ T cells. CD4+CD25highFOXP3+ as well as CD4+CD25highFOXP3+ST2+ Tregs were analyzed by flow cytometry. In a group of patients with type 1 diabetes in vitro IL-33 treatment induced regulatory CD4+CD25highFOXP3+ cell frequencies as well as upregulating the surface expression of ST2 molecule. In addition, the number of CD4+CD25highFOXP3+ cells carrying ST2 receptor increased significantly. Similar effect was observed in case of the FOXP3 expression. We did not observe any significant changes in IL-33 treated cells of healthy controls. The level of ST2 was higher in serum of patients with type 1 diabetes in comparison to their healthy counterparts. We propose that IL-33 becomes an additional immunostimulatory factor used to induce Treg expansion in future clinical trials of adoptive therapy in type 1 diabetes.

Quantitative and functional characteristics of endothelial progenitor cells in newly diagnosed hypertensive patients

Populations of peripheral blood CD34+ cells comprise precursors of endothelial cells. These precursors are crucial to cardiovascular homeostasis. Hypertension, as one of the main risk factors for cardiovascular disease, is associated with the loss of endothelium structural integrity and its functional impairment. The aim of our study was to evaluate the subsets of endothelial precursor cells in patients with newly diagnosed arterial hypertension. Twenty-four newly diagnosed, previously untreated hypertensive patients aged 59.5±12.5 years, were enrolled into the study group, whereas the control group comprised 45 healthy subjects, 55.5±10.0 years old. Endothelial progenitor cells (EPCs) were analysed by flow cytometry. The results showed that hypertensive patients were characterized by a significantly higher percentage and number of the CD34+ cells and simultaneously less differentiated CD34+CD45dim/negCD133+ progenitors. The percentage and number of CD34+CD45negVEGFR2+ and CD34+CD45negCD133+VEGFR2+ cells were not different from the control group. Moreover, patients had a significantly lower percentage and number of the CD34+CD45negVEGFR2+CXCR4+ and CD34+CD45negVEGFR2+ICAM-1+ cells than healthy individuals. These changes were paralleled by early symptoms of nephropathy, that is, lower glomerular filtration rate (GFR) values and borderline micro albuminuria. Our results indicate that an elevation in the number of less differentiated progenitors may be a mechanism compensating for defects of migration and adhesion, present in a more differentiated subset.

CCR5-Δ32 gene polymorphism is associated with retinopathy in patients with type 1 diabetes.

AIM The aim of the study was to assess the relationship between the CCR5-Δ32 polymorphism and the risk of diabetic retinopathy (DR) in patients with DM1. METHODS We examined 420 patients and 350 healthy controls. The analysis concerned CCR5-Δ32 polymorphism as well as levels of serum inflammatory markers (CRP, TNF-α), adhesion molecules (VCAM, ICAM-1, ICAM-3) and CCR5 ligand (MCP-1). RESULTS We found a negative association between DM1 and Δ32 allele. Moreover, the frequency of Δ32 allele was higher in a group with DR in comparison to control subjects without this complication. We also found that Δ32 carriers had the highest levels of: HbA1c, inflammatory markers, adhesion molecules and CCR5 ligand. CONCLUSIONS The findings of our studies suggest that the CCR5-Δ32 polymorphism is associated with DM1 such that the Δ32 allele protects against the development of DM1 and increases the risk of DR in patients who have already developed the disease.

Estrogen receptor α gene polymorphism and vascular complications in girls with type 1 diabetes mellitus

The effect of estrogens is mediated by activation of estrogen receptors (ERs). Because ER-α gene polymorphisms may exert different effects in childhood, we analyzed the associations between the IVS1 −397T>C (PvuII) polymorphism and systemic inflammatory state, proangiogenic factors, frequency of monocyte subsets, lipid profile, blood pressure, and vascular complications in girls with type 1 diabetes mellitus (DM1). We examined 180 young girls with DM1 and 120 healthy age-matched controls. The analysis concerned PvuII polymorphism of the ER-α gene as well as the levels of serum inflammatory markers (CRP, IL-6, TNF-α), proangiogenic factors (VEGF, angiogenin), 17β-estradiol, values of monocyte subsets (CD14++CD16− and CD14+CD16+), lipid profile, and blood pressure. In our study, girls with CC genotype had lower level of inflammatory and angiogenic factors and lower frequencies of CD14+CD16+ monocytes in comparison to CT or TT carriers. Simultaneously, the CC carriers had a greater population of CD14++CD16− monocytes, increased blood pressure, and serum levels of: estrogen, total cholesterol, triglycerides, and low-density lipoprotein cholesterol than girls bearing CT or TT genotype. Our study suggests a pleiotropic effect of PvuII polymorphic CC variant on diabetic vasculopathies. Although the CC genotype carriers demonstrate less inflammatory and angiogenic activity, they seem to display less favorable cardiometabolic features. Based on our study, we cannot distinguish PvuII ER-α genotype that could be useful in identification of DM1 girls that are more prone to develop of late vascular complications, before the occurrence of first clinical symptoms.

Association of circulating progenitor cells with angiotensin II in newly diagnosed hypertensive patients

Populations of CD34− and VEGFR2-expressing cells are responsible for regeneration of damaged endothelium and vascular remodelling. As their quantity and activity changes during cardiovascular diseases, they are potentially useful markers of cardiovascular health. The aim of our study was to investigate changes of various CD34+ and CD34+ VEGFR2+ populations in subjects with newly recognised hypertension and to evaluate whether observed alterations are influenced by clinical parameters and angiotensin II. Circulating CD34+ and CD34+ VEGFR2+ cells were analysed in peripheral blood samples by flow cytometry. Serum levels of angiotensin II were determined using immunoenzymatic assay. We discovered increased proportions of various CD34+ populations and CD34+ VEGFR2+ c-Kit+ cells in newly diagnosed patients. CD34+ cells seem to be influenced by angiotensin II, but we did not observe comparable results when populations co-expressing VEGFR2 were analysed. The quantity of CD34+ VEGFR2+ cells in patients with newly recognised primary hypertension ought to be determined by other factors. Increased proportions of CD34+ progenitors in blood could comprise compensatory mechanism for increased endothelial damage in hypertension.

Sex-related association of serum uric acid with inflammation, kidney function and blood pressure in type 1 diabetic patients

Recent studies suggest that uric acid (UA) is a mediator of diabetic nephropathy. We hypothesized that serum UA would associate with the prevalence of diabetic nephropathy in youth with type 1 diabetes (T1D), and that this relationship would differ by sex.

CCR5-Δ32 gene polymorphism is related to celiac disease and autoimmune thyroiditis coincidence in patients with type 1 diabetes

AIM The aim of the study was to assess the relationship between CCR5-Δ32 polymorphism and the coincidence of celiac and autoimmune thyroid diseases with type 1 diabetes mellitus (T1D) in children. METHODS 420 children with T1D aged 15.5±3.0years and 350 healthy controls were studied. Characterization of CCR5-Δ32 genotypes (rs333) was analyzed by polymerase chain reaction (PCR). RESULTS The allele frequency was significantly different in diabetic children as compared to the healthy controls (p<0.0001). We found negative association between T1D and Δ32 allele (OR=0.383; 95% CI=0.268-0.549). Besides, we observed alterations in the frequencies of CCR5-Δ32 genotypes due to celiac and autoimmune thyroid diseases. The risk of celiac disease for patient carriers of the 32-bp deletion was more than threefold higher than for noncarriers (OR=3.490; 95% CI=1.357-8.859; p=0.009). Similar results were obtained in the case of autoimmune thyroiditis. The risk of autoimmune thyroiditis for patient carriers of the 32-bp deletion was also more than threefold higher than for noncarriers (OR=3.466; 95% CI=1.754-6.849; p=0.0004). CONCLUSIONS The findings of our studies suggest that the CCR5-Δ32 polymorphism is associated with type 1 diabetes mellitus and the Δ32 allele increases the risk of celiac disease and autoimmune thyroid disorders in patients with T1D.

The GG genotype of the −152 G/A vascular endothelial growth factor (VEGF) polymorphism predisposes to hypertension-related chronic kidney disease

Our purpose was to determine whether the VEGF −152 G/A polymorphism could be associated with chronic kidney disease and endothelial dysfunction in hypertensive patients. There were 100 healthy volunteers enrolled into the control group. The group of patients was constituted by 99 consecutively admitted hypertensive patients referred to our Institution by their general practitioner. All patients were treated with anti-hypertensive polytherapy. Presented study revealed that the hypertensive patients bearing the GG genotype were characterized by the highest values of diastolic blood pressure and markers of endothelial damage such as Angiogenin, Endostatin, CRP as well as von Willebrandt factor. In addition, higher number of immature endothelial progenitor cells with CD34+CD133+, CD34+CD133- markers was observed in GG hypertensive carriers while in normotensive individuals no differences were found. Such phenomenon may indicate an increased mobilization of bone-marrow derived endothelial progenitors. It may testify to the preserved compensatory mechanism in chronic kidney disease (CKD) patients until the G3a stage of the disease. Moreover, patients with higher estimated glomerular filtration rate (eGFR) level had lower of vWf and Endostatin values, and higher level of VEGF. Taken together our findings clearly indicate the −152 GG hypertensive carriers as more prone to develop CKD. We can suspect that the VEGF −152 GG genotype is strongly associated with hypertension-dependent CKD.