Antoni Hrycek

Long pentraxin 3 (PTX3) in the light of its structure, mechanism of action and clinical implications

Pentraxins are a group of evolutionarily conserved ancient proteins. Depending on their structure, pentraxins are divided into short and long pentraxin families. Pentraxin 3 (PTX3) is the prototype of the long pentraxin group. PTX3 synthesis is stimulated by a variety of molecules involved in the inflammatory process. The inflammatory mediator is typically produced at inflammatory sites; however, it can also be released at the sites remote from the original inflammatory insult. Although mainly expressed by vascular endothelium and smooth muscle cells, PTX3 is also synthesized by myeloid dendritic cells, mononuclear macrophages/phagocytes, vascular endothelial and smooth muscle cells, fibroblasts, adipocytes, cumulus oophorus cells mesangial cells, synovial cells and chondrocytes. PTX3 binds to several ligands including complement component C1q, factor H, ficolin-1 (M-ficolin), mannose-binding lectin, fibroblast growth factor 2, P-selectin, matrix protein TSG6 and Klebsiella pneumoniae; it is also known to play a role in humoral innate immunity as well as in degenerated and apoptotic cells clearance. PTX3 acts as a modulator of inflammatory processes, modifies angiogenesis and atherosclerotic lesion development, and participates in extracellular matrix formation. Due to the fact of PTX3 being primarily produced and released by vascular wall cells, it might be used as a sensitive and independent inflammatory marker.

HLA-DRB1 and -DQB1 alleles and gene polymorphisms of selected cytokines in systemic lupus erythematosus

ObjectiveThe objective of this study was to evaluate the genetic profiles of selected cytokines (transforming growth factor beta 1, tumor necrosis factor alpha, interleukin-6, interferon gamma, and interleukin-10) in systemic lupus erythematosus and the contributions of human leukocyte antigen (HLA)-DRB1 and -DQB1 alleles to susceptibility for this disease.Patients and methodsThe study was carried out in 24 SLE patients and 36 healthy controls (from Upper Silesia) using polymerase chain reaction methods. All persons were of Caucasoid origin. Standard association analysis was used to compare the HLA alleles and frequency of cytokine gene polymorphisms between these groups.ResultsOnly the frequency of HLA-DRB1*07 allele was higher in SLE patients than controls (odds ratio 2.92, 95% confidence interval 1.16–7.33), but the difference did not reach statistical significance when Bonferroni’s adjustment procedure was performed. No other significant associations were noted between class II alleles (DR1-DR6, DR8-DR10, DQ1-DQ4) and SLE. The frequency of the interleukin-6 GG and GC genotypes was significantly higher in SLE patients than in controls, and a significantly higher percentage of the G vs C alleles between patients and controls was revealed (odds ratio 2.53, 95% confidence interval 1.37–4.65, chi-squared test 8.16, P<0.05). The most significant association of increased frequency of the G allele with SLE was more commonly noted in HLA-DRB1*07-positive patients (odds ratio 10.29, 95% confidence interval 5.34–19.83, P<0.001). These data indicate that this combination could contribute toward determining the susceptibility to SLE, but its possible significance will require confirmation by further studies.

Human cytomegalovirus in patients with systemic lupus erythematosus

The objective of this study was to determine the frequencies of human cytomegalovirus (HCMV) infection and HCMV genome copy number in blood of consecutive (treated from several months to several years) systemic lupus erythematosus (SLE) patients (22 women). The obtained results were compared to the healthy controls (15 women). All patients fulfilled at least four of the 1982 revised American rheumatism association (ARA) classification criteria for SLE. Our patients demonstrated three or four of the nine possible organ systems involved and most of them had mild SLE with SLE disease activity index (SLEDAI) score < 10 at time when blood samples were collected to detect HCMV. Quantitative analysis of HCMV genome was performed with aid of sequence analyzer ABI PRISM TM 7700 Perkin Elmer. Primers and probe were constructed on the basis of IE4 region of HCMV genome. The viral load was expressed as log10 of calculated HCMV genome copy number. Qualitative analysis revealed that 100% of our SLE patients were infected with HCMV, whereas in the control group only 73% of persons were HCMV positive. Statistically significant difference was demonstrated when the strength of the association between SLE or controls and infection of HCMV was calculated (estimated by Fishers exact test, P value = 0.02). Higher viral DNA copy number was observed in whole blood of SLE patients than in the control group (338.45 ± 221.76 and 229.00 ±405.61 copies/ml respectively) but did not reach statistical significance level (95% confidence interval from 170.41 to 249.32, P = 0.71). Furthermore percentage of patients with HCMV-DNA copy number >2.0 × 102 copies/ml was statistically significantly higher than this one in controls. The data show association between HCMV infection and SLE, which should be taken into account during the course of SLE.

A Comparative Clinical Study on Five Types of Compression Therapy in Patients with Venous Leg Ulcers

The aim of this study was to compare five types of compression therapy in venous leg ulcers (intermittent pneumatic vs. stockings vs. multi layer vs. two layer short stretch bandages vs. Unna boots). Primary study endpoints were analysis of changes of the total ulcer surface area, volume and linear dimensions inside observed groups. The secondary end points were comparisons between all groups the number of completely healed wounds (ulcer healing rates), Gilman index and percentage change of ulcer surface area. In total, 147 patients with unilateral venous leg ulcers were included to this study. Participants were randomly allocated to the groups: A, B, C, D and E. After two months the healing rate was the highest in group A (intermittent pneumatic compression) - 57.14%, 16/28 patients, B (ulcer stocking system) - 56.66%, 17/30 patients and C (multi layer short stretch bandage) - 58.62%, 17/29 patients. Significantly much worse rate found in group D (two layer short stretch bandages) - only 16.66%, 5/30 patients and E (Unna boots) - 20%, 6/30 patients. The analysis of changes of the percentage of Gilman index and wound total surface area confirmed that intermittent pneumatic compression, stockings and multi layer bandages are the most efficient. The two layer short - stretch bandages and Unna boots appeared again much less effective.

Pentraxin 3 as a biomarker of local inflammatory response to vascular injury in systemic lupus erythematosus

Abstract Systemic lupus erythematosus (SLE) is an autoimmune disorder with organ injury related to vasculitis. Inflammation of blood vessels results from auto-immunological activation of endothelial cells. The pentraxin 3 (PTX3), might act as an indicator of vasculitides in many diseases. The aim of this study was to determine whether PTX3 might be useful as a marker of vascular injury in SLE. This study was carried out in a group of 56 SLE women, and in the 28 female volunteers control group. All participants’ plasma and serum samples were collected to estimate concentrations (ELISA) of PTX3, soluble thrombomodulin, soluble E-selectin (sE-selectin), soluble P-selectin (sP-selectin), soluble form of vascular cell adhesion molecule 1 (sVCAM-1), soluble inter-cellular adhesion molecule-1 (sICAM-1), soluble platelet endothelial cell adhesion molecule 1, monocyte chemotactic protein-1 (MCP-1) and von Willebrand factor (vWF) activity. Anthropometric, demographic and lifestyle characteristics of SLE patients were also performed. The SLE patients had higher PTX3, vWF, MCP-1, sE-selectin and sVCAM-1 levels than the controls (1.82 ± 1.56 ng/mL, 237 ± 101%, 70.05 ± 18.31 ng/mL, 111.16 ± 49.15 ng/mL and 978.78 ± 462.35 ng/mL vs. 0.86 ± 0.40 ng/mL, 138 ± 43%, 58.56 ± 13.91 ng/mL, 66.04 ± 27.18 ng/mL and 499.07 ± 125.67 ng/mL, respectively). The independent factors affecting PTX3 expression included Systemic Lupus Erythematosus Disease Activity Index, prednisone dose and anemia severity. Moreover, the PTX3 areas under the curve–receiver operating characteristics curves 0.717 ± 0.056 with cut-off level of 1.96 ng/mL was comparable to vWF, MCP-1, sE-selectin, sP-selectin and sICAM-1. PTX3 and sVCAM-1 were the only factors related to SLE activity. Other vascular injury indicators associated with PTX3 were vWF and sVCAM-1. In conclusion, PTX3 concentrations in SLE patients might serve as a indicator of the activation/dysfunction of vascular endothelium.

Selected acute phase proteins and interleukin-6 in systemic lupus erythematosus patients treated with low doses of quinagolide

The relationship between endocrine regulation and immune system has recently become the subject of intense investigations. The objective of this study was to determine the extent of selected serum acute phase proteins (APP), IL-6 and Systemic Lupus Erythematosus Disease Activity Index (SLEDAI) involvement in systemic lupus erythematosus (SLE) patients during quinagolide therapy. A further aim of this study was to evaluate the relationships between the above mentioned parameters. In 25 SLE patients treated with a low dose of quinagolide (12.5–50 μg per day) and in 25 healthy persons who constituted the control group, serum concentration of C-creative protein (CRP), alpha-1-antitripsin (AAT), ceruloplasmin (CER), IL-6 and prolactin (PRL) were estimated at entry and in patients after 3 months of treatment. Moreover, SLEDAI score was calculated at entry and after 3 months of therapy with quinagolide. IL-6 and PRL levels were significantly higher in SLE group whereas the concentrations of CRP, AAT and CER were higher than in the controls, but without statistical significance. After 3 month therapy statistically significant decrease of serum level of IL-6 and PRL was revealed. Statistically significant lower serum concentration of CER was also obtained after 3 months of therapy whereas serum CRP and AAT concentration was lower compared with the mean pretreatment level but the results did not reach statistical significance. A raised SLEDAI score at entry was significantly reduced after 3 month therapy and positive correlation with PRL level in examined group of patients with SLE was noted at entry. The decreased serum concentration of IL-6, APP and SLEDAI score observed during applied therapy with small dose of quinagolide confirms the hypothesis that quinagolide may become a valuable and safe drug in the therapy of patients with mild SLE.

Serum levels of selected chemokines in systemic lupus erythematosus patients

Chemokines promote leukocyte traffic into the site of inflammation. Serum levels of monocyte chemotactic protein 1 (MCP-1), stromal cell-derived factor-1 (SDF-1), interferon-gamma-inducible protein 10 (IP-10), and interleukin-8 (IL-8) were evaluated in 48 treated women with systemic lupus erythematosus (SLE) and mild-to-moderate disease severity. The results were compared between the whole SLE group and the control (29 women). The relationships between chemokines, their concentrations, and peripheral blood leukocyte count and between the chemokines and individual leukocyte populations (polymorphonuclear leukocytes—PMNs, lymphocytes—Ls, monocytes—Ms, eosinophils) counts were determined. The relationships between the analyzed chemokines were also determined in the control. SLE subjects had significantly higher MCP-1, SDF-1, IP-10, and lower IL-8 concentrations compared to the control. Moderate, positive correlations between MCP-1/SDF-1, SDF-1/IP-10 and a negative correlation between MCP-1/IL8 were observed in the patient group. Moderate, negative correlations were found between SDF-1/total leukocyte count, SDF-1/absolute number of PMNs as well as between IP-10/total leukocyte count, IP-10/absolute PMNs, Ls, and Ms counts in peripheral blood of SLE group. We suggest that the obtained results and correlations observed between the examined parameters might be used to monitor SLE course and progression. However, further randomized clinical studies should be carried out on in untreated and treated patients with SLE.

Coeliac disease in systemic lupus erythematosus: a case report

The case of a rare coexistence of systemic lupus erythematosus (SLE) with coeliac disease (CD) is described. Systemic lupus erythematosus was diagnosed prior to CD and initially SLE treatment was administered. After several years, when CD symptoms developed, the diagnosis was corrected and additional treatment with a gluten-free diet was applied with beneficial effects.

Annexin A5 and anti-annexin antibodies in patients with systemic lupus erythematosus

Plasma levels of annexin A5 (ANX A5) and anti-annexin A5 (aANX A5) antibodies were evaluated in 51 women with systemic lupus erythematosus (SLE). The results were compared between the total SLE group, subgroups on/without immunosuppressive therapy and the control (28 women). The relationships between ANX A5/aANX A5 antibodies levels and laboratory variables (anti-cardiolipin antibodies—aCL, total cholesterol, thrombocyte count, activated partial thromboplastin time—APTT, prothrombin time, international normalized ratio–INR) were performed in the total SLE group and in the patient subgroups identified as the arithmetic mean of ANX A5 concentration in the control plus 1–4 standard deviations (SD). The whole SLE group and the subgroup on immunosuppression showed significantly higher ANX A5 and IgG aANX A5 antibodies concentrations. A weak positive correlation was found between ANX A5 and thrombocyte count, a moderate one between IgG and IgM aANX A5 antibodies, a weak negative correlation between IgG aANX A5 and APTT in the whole SLE group. SLE subgroups with ANX A5 concentrations higher than the control mean plus 3 or 4 SD showed a weak/moderate negative correlation of this parameter with aANX A5 antibodies, moderate one with IgG aCL antibodies levels, a moderate positive correlation with cholesterol concentration, moderate/high positive correlations with thrombocyte count. The association between plasma ANX A5/IgG aANX A5 levels and severity of disease was noticed. The role of aANX A5 and IgG aCL antibodies as causative factors of increased ANX A5 levels was suggested, and the relationship between ANX A5 and thrombocyte count was revealed.

Peripheral blood lymphocytes and selected serum interleukins in workers operating X-ray equipment

The objective of these studies was to determine the effect of low X-ray doses on peripheral blood lymphocytes in the workers operating X-ray equipment. In 30 workers and 18 persons who constituted the control group the absolute number of peripheral blood lymphocytes, the number of T CD4+ and T CD8+ lymphocytes, the ratio CD4+/CD8+ and the concentration of serum interleukins (IL-2, IL-4, IL-6) were tested. Subgroups with respect to sex and employment period were selected. A significant increase in IL-2 concentration in workers as compared to controls was observed and significantly decreased IL-4 level in operating X-ray equipment was revealed. The lowest levels of IL-4 were noted in the women subgroup. A high positive, statistically significant correlations between the number of lymphocytes and the concentration of IL-2 and the level of IL-4 were observed. Moreover, our studies revealed that lymphocyte number and interleukin concentrations in serum of X-ray operators do not depend on the assumed ranges of the length of employment period.