Glucocorticoids upregulate decreased IL-7 receptor expression in asthmatic patients and simian immunodeficiency virus-infected non-human primates.

Signaling through interleukin-7 receptor (IL-7R) is essential for regulation of T-cell homeostasis and survival. Previously, we and others have found diminished IL-7R levels in simian immunodeficiency virus (SIV) - infected non-human primates and human immunodeficiency virus (HIV) - infected patients. To date, it remains unknown whether changes in IL-7R expression could also be linked to non-infectious inflammatory diseases such as asthma or anti-inflammatory drug use. Here, we investigated through flow cytometry the levels of IL-7R expression on CD4+ and CD4- T-cells in asthmatic patients in relation to disease severity, immune status and glucocorticoid (GC) use. In addition, we sought to evaluate the effects of in vivo and in vitro GC treatment on IL-7R expression in both asthmatic patients and SIV-infected non-human primates. We demonstrated that expression of IL-7R on peripheral blood CD4+ T-cells was significantly decreased in clinically stable GC-naive mild and moderate asthmatic patients. Accordingly, the development of asthmatic reaction following bronchial allergen challenge performed in sensitized subjects was associated with a significant drop in levels of IL-7R on circulating CD4+ T-cells. In contrast, CD4+ T-cells from both, mild and moderate, but not severe asthmatics, treated with inhaled GC displayed levels of IL-7R similar to that seen in healthy controls. We did not find significant differences with serum or sputum interleukin-7 levels among healthy controls and GC-naïve and GC-treated asthmatic patients. Furthermore, both in vitro GC treatment and short-term oral GC administration to asthmatic patients resulted in a significant enhancement of IL-7R. Similarly, we demonstrated that GC-stimulated T-cells from SIV-infected non-human primates up-regulated IL-7R expression. Accordingly, experimental short-term systemic in vivo administration of GC to SIV-infected macaques led to enhancement of IL-7R expression on circulating T-cells. Our data indicate that GC bear potential to recover diminished IL-7R expression, as well in asthma as in lentiviral infection.

Generation of T regulatory cells in children with newly diagnosed type 1 diabetes mellitus.

UNLABELLED: There is increasing evidence that T-regulatory (Treg) cells could be used to prevent or cure autoimmune diseases including type 1 diabetes mellitus (T1DM). The aim of the present study was to verify the hypothesis that functional Treg cells can be generated from conventional T-cells separated from a small amount of peripheral blood of children with newly diagnosed T1DM (N=25). METHODS: CD4(+)CD25(-) cells were cultured with Treg expander (CD3/CD28) and IL-2 for generating de novo Treg cells. The assessment of the expression of selected genes and proteins critical to Treg function and the proliferation assays were performed with the use of real-time RT-PCR and flow cytometry. RESULTS: After a 4-week stimulation with Treg expander and IL-2, the percentage of T-regulatory cells was significantly higher compared to the cells treated with medium alone (with no difference between diabetic and control children). However, we found some disturbances in the gene expression at mRNA level for molecules crucial for T-reg function. The induced Tregs from diabetic and control children were fully functional as assessed in proliferation assays. IN SUMMARY: Despite some disturbances at mRNA level in the critical gene expression, the suppressive properties of induced Treg cells from diabetic and control children were effective.

Overexpression of B cell-activating factor (BAFF) in neutrophils of oral cavity cancer patients - preliminary study.

In the present study the expression of tumor-promoting B cell-activating factor (BAFF), a member of the tumor necrosis factor superfamily (TNF) in neutrophils from oral cavity cancer patients, was examined by real-time PCR. For the purpose of comparison, the expression of BAFF protein was assessed in autologous peripheral blood mononuclear cells (PBMCs). An important question of this study has also been to explain the role of NF-κB in the induction of BAFF molecule. The increased expression of BAFF at the mRNA and protein levels in neutrophils and mononuclear cells of patients before and after treatment were accompanied by the increased expression of phospho-IκB protein level. Demonstrated excessive expression and secretion of BAFF by examined leukocytes suggest a tumor-promoting activity of those cells in oral cavity cancer patients. The overexpression of BAFF, observed at mRNA and protein levels in PMNs and PBMCs, as well as the secretion of soluble form of sBAFF by these cells, accompany the increased concentrations of sBAFF in the serum of patients. Observations above suggest that the modulation of BAFF molecules in examined leukocytes and the levels thereof in the serum may have future implications for immunotherapy of oral cavity cancer patients.

Disturbances in some gene expression in T regulatory cells separated from children with metabolic syndrome.

The metabolic syndrome (MS) is defined as a cluster of risk factors, including abdominal obesity, dyslipidaemia, glucose intolerance and hypertension, which increase the risk for coronary heart disease. The immunological aspects of obesity and MS, including the role of T regulatory cells, have been intensively investigated. The aim of this study was to determine whether there is any disturbance in T regulatory cells number and/or function in children with MS. The percentages of T regulatory cells in the peripheral blood of children fulfilling the International Diabetes Federation criteria of the disease (n = 47) were assessed. Treg cells were also separated for further analysis of multiple genes important in their function with the use of real-time RT-PCR. We did not observe any difference in Treg percentages between study and control group but there was lower expression of some molecules including transforming growth factor-beta and interleukin-12 family members in Treg cells separated from children with MS compared to the healthy subjects. Our study is the first to report significant disturbances in some gene expression in T regulatory cells separated from children with MS. The results should be useful for further research in this field, including immunotherapeutic interventions.

Ghrelin in gestational diabetes: serum level and mRNA expression in fat and placental tissue.

OBJECTIVES: We studied the effect of an oral glucose load on circulating ghrelin, as well as ghrelin and ghrelin receptor (GHS-R1a) mRNA expression in subcutaneous adipose tissue (SAT), visceral adipose tissue (VAT) and placental tissue from pregnant women with gestational diabetes (GDM) and normal glucose tolerance (NGT). METHODS: Plasma total ghrelin levels were measured in 58 patients with GDM and 61 women with NGT by radioimmunoassay. Ghrelin and GHS-R1a mRNA expression was studied in 16 subjects with GDM and 20 healthy pregnant women at term, using RT-PCR. RESULTS: Basal ghrelin concentrations and the maximal decrease in ghrelin levels after glucose load did not differ in the women with GDM and NGT (399.1 [299.6-563.3] pg/ml vs. 400.9 [302.3-475.8] pg/ml and 127.6 [23.1-213.1] pg/ml vs. 101.7 [44.0-217.6] pg/ml, respectively). Ghrelin mRNA expression in placental tissue was significantly higher in the subjects with GDM than in the healthy pregnant women (0.06 [0.03-0.07] AU vs. 0.02 [0.015-0.03 AU], p=0.02), whereas GHS-R1a mRNA expression in all three tissues studied did not differ between the two groups. Multiple regression analysis revealed that ghrelin mRNA expression in SAT was significantly predicted by serum insulin (beta=0.62, p=0.01), explaining 42% of its variability. CONCLUSIONS: Ghrelin mRNA expression in placental tissue was higher in the GDM than in NGT subjects, whereas no association between circulating ghrelin and GDM was observed.

The FTO gene modifies weight, fat mass and insulin sensitivity in women with polycystic ovary syndrome, where its role may be larger than in other phenotypes.

AIM: Genome-wide association studies have shown that variation in the FTO gene predisposes to obesity and related traits that are common features of polycystic ovary syndrome (PCOS). The aim of the present study was to assess the effect of FTO variation on obesity, insulin sensitivity, and metabolic and hormonal profiles in PCOS. METHODS: We examined 136 PCOS women (mean body mass index [BMI]: 28.28+/-6.95kg/m(2), mean age: 25.36+/-5.48 years). Anthropometric measurement, euglycaemic-hyperinsulinaemic clamp and oral glucose tolerance tests and sex hormone assessments were performed. The study group was genotyped for the FTO rs9939609 polymorphism. RESULTS: BMI (29.0+/-6.9kg/m(2) vs 26.1+/-6.8kg/m(2); P=0.023), body weight (80.1+/-20.7kg vs 72.6+/-20.2kg; P=0.048), fat mass (29.7+/-1 6.6kg vs 24.6+/-17.7kg; P=0.045) and waist circumference (89.8+/-16.7cm vs 83.2+/-17.1cm; P=0.028) were higher in carriers of at least one copy of the A allele. Differences in these parameters were more significant when comparing AA and TT homozygotes. Women with the AA genotype also had decreased insulin sensitivity (P=0.025) and follicle-stimulating hormone (P=0.036). In logistic-regression analyses, the association of the FTO gene polymorphism with insulin sensitivity was no longer significant when BMI was included in the model. CONCLUSION: Variation in the FTO gene modifies weight, adiposity and other measures of obesity and insulin sensitivity in PCOS. The examined FTO gene variant appears to have a greater impact on obesity and related traits in PCOS than in other phenotypes. The effect on insulin sensitivity appears to be secondary to its influence on obesity and body fat.

Single Nucleotide Polymorphisms in exon 3 of the adiponectin gene in subjects with type 2 diabetes mellitus.

PURPOSE: Adiponectin (APM1)--a newly discovered adipocytokine secreted by fat tissue--was recently suggested to play a role in the genetic predisposition to type 2 diabetes, obesity and insulin resistance. Adiponectin gene is localized on chromosome 3q27 within the region which was identified as susceptibility locus for type 2 diabetes and metabolic syndrome. Till now genetic associations of two SNP in exon 2 (+45T/G) and intron 2 (+276G/T) of adiponectin gene with type 2 diabetes and adiponectin level were reported in Japanese population and with insulin resistance in some Caucasian populations (Italy, Germany). Moreover, in the proximal promoter region of the APM1 gene: SNP-11426A/G and -11391A/-11377G haplotype predicted the associations with fasting plasma glucose, type 2 diabetes and adiponectin levels. On the other hand the role of mutations in exon 3 of the adiponectin gene is not so well studied. MATERIAL AND METHODS: The aim of our study was the screening for rare mutation in exon 3 of adiponectin gene in the Polish subjects with type 2 diabetes as there is no data available about the frequency and role of these mutations in our population. The study was performed in the group of 187 Polish origin patients with type 2 diabetes (32 female and 155 male, mean age 54.1 +/- 8.6 yrs) and 102 age and sex matched healthy controls. RESULTS: The frequency of adiponectin gene mutations in exon 3 was 3.9%, while in the control group 0.98% and this difference was not statistically significant. We also observed that adiponectin level is significantly lower in patients with c.331 T-->C mutation (Y111H) in comparison to subjects without this mutation (5.0 ug/ml vs 14.4 ug/ml, p=0.0148). CONCLUSIONS: To our knowledge the present study is the first which shows that in Polish populations.

Interleukin 18 and sICAM-1 serum levels in families with type 1 diabetes mellitus.

It is well known that subjects with type 1 diabetes are at an increased risk of death from coronary heart disease in comparison to non-diabetic age-matched individuals because hyperglycaemia is believed to be a key risk factor for the development of micro- and macrovascular complications. On the other hand there is increasing evidence about the role of inflammatory mediators in the pathogenesis of atherosclerosis and the development of acute coronary syndromes. It has been recently suggested that IL-18 and sICAM-1 have a strong predictive value for cardiovascular diseases and deaths in patients with coronary artery disease and/or in apparently healthy men. The aim of our study was to estimate the serum levels of IL-18 and sICAM-1 in subjects with type 1 diabetes and their relatives, who share HLA diabetic susceptibility genes (with or without pancreatic autoantibodies), but still without glucose level disturbances, as an evaluation of the possible role of genetic predisposition to the presence of IL-18 in diabetic families. The study was carried out in 35 type 1 diabetic subjects, their 101 healthy first-degree relatives: 36 siblings and 65 parents and the control group consisted of 31 healthy volunteers. We have found increased IL-18 and sICAM-1 levels in subjects with type 1 diabetes and their first degree relatives, who share diabetic HLA haplotypes: DRB1*03/DRB1*04 or DRB1*03/*04/DQB1*02 independently of their autoimmune status. There was a strong positive correlation between IL-18 and sICAM-1 levels in diabetic subjects and their first degree relatives without glucose level disturbances. To our knowledge this is the first study, which suggests that sICAM-1 elevations could be a result of IL-18 overproduction in type 1 diabetic subjects and their first degree relatives. Since in previous studies IL-18 and sICAM-1 were found to be predictors of death in subjects with CHD, one could speculate that high levels of IL-18 observed in subjects with genetic predisposition, but still with normal glucose levels, are an in addition to hyperglycaemia, pathogenic factors responsible for a higher risk of acute coronary events in subjects with diabetes type 1.