Joanna Osada

Expression of interleukin 4 receptors in bronchial asthma patients who underwent specific immunotherapy.

BACKGROUND Interleukin (IL) 4 and IL-13 are crucial cytokines for the development of allergic reactions and have been shown to modulate the function of monocytes and macrophages. OBJECTIVES To evaluate the expression of IL-4Rs on peripheral blood monocytes and in the serum of patients with bronchial asthma who underwent specific immunotherapy (SIT). METHODS The study was performed on 17 asthma patients with a typical clinical history and positive skin prick test results to Dermatophagoides pteronyssinus allergens. Five asthma patients who declined SIT were used as a comparator control group. Ten healthy persons served as negative controls. Flow cytometry analysis was performed on the whole blood samples using labeled monoclonal antibodies against CD14 and CD36 monocyte markers and against the CD124 alpha chain of IL-4R. The serum levels of soluble IL-4R were evaluated using an immunoenzymatic assay. RESULTS Compared with controls, bronchial asthma patients before SIT had a higher mean +/- SD percentage of CD14-positive cells that coexpressed CD124 (3.5% +/- 1.8% vs 18.6% +/- 7.9%; P < .01). After SIT, the mean +/- SD percentage of CD14 cells coexpressing CD124 decreased to 8.1% +/- 5.1%, which was significantly lower than before SIT (P < .01) but still significantly higher than in controls (P = .01). Changes in CD124 expression were associated with up-regulation of CD14 and down-regulation of CD36 expression on peripheral blood monocytes, suggesting that IL-4/IL-13-mediated signaling may be important for regulation of monocyte phenotype and function in asthma patients receiving SIT. CONCLUSIONS Even short courses of SIT are associated with a decrease in IL-4R expression on peripheral blood monocytes, which may cause decreased IL-4/IL-13-mediated effects in patients who undergo SIT.

Monocyte Subsets and Natural Killer Cells in Acute Pancreatitis

Background: Alteration of the immune system is one of the major mechanisms responsible for complications in severe acute pancreatitis (AP). The aim of our study was to provide a complex evaluation of peripheral blood monocyte subsets, natural killer cells (NK cells) and cytotoxic T lymphocytes in patients with different severity forms of AP. Methods: 20 patients with mild AP and 15 with severe AP (S-AP) were included in our study. Peripheral blood mononuclear cells were studied on days 1–3, 5, 10 and 30, by means of flow cytometry. Results: In peripheral blood of patients with pancreatitis, we found a marked increase in total monocyte count. In S-AP, circulating monocytes were significantly activated, which was presumed from increased expression of HLA-DR, CD54, CD69 and CD25. Concurrent increased expression of CD95 (FasR) may indicate enhanced susceptibility of these cells to apoptosis. In patients with S-AP, a dramatic depletion of circulating NK cells (CD16/56 and CD3– CD8+) was found along with a reduction of circulating CD3+ CD8+ lymphocytes (cytotoxic T lymphocytes). Conclusion: Our findings suggest profound disturbances of innate cellular immunity in patients with S-AP.

Does prematurity affect platelet indices

PURPOSE The current study objective was to compare blood platelet indices in preterm newborns (PTN) and full term newborns (FTN). MATERIALS AND METHODS We introduced to our study 51 PTN (25 females, 26 males) and 55 FTN (25 females, 30 males). Platelet indices were estimated in blood samples collected from the umbilical artery. RESULTS PTN demonstrated a decreased count of blood platelets (197 x 103/microL) as compared to FTN (287 x 103/microL), p=0.0001. Platelet hematocrit (PCT) also showed substantial differences in both groups (PTN=0.16% vs. FTN=0.22%; p=0.001). Mean platelet volume (MPV) was found to be nearly the same (PTN=8.02 fl, FTN=7.79 fl). Platelet distribution width (PDW) was higher in PTN (50.64%) than in FTN (46.54%), p=0.021. Large platelet count (LPLT) was diminished in PTN (5.23%) in comparison with FTN (6.12 %). CONCLUSIONS A decreased count of blood platelets, platelet hematocrit and increased platelet distribution width may result from a low gestational age or a dysfunction of megakaryocytes and the placenta. Blood platelet indices may be vital in the diagnosis of haemostatic disorders.

The expression of the chemokine receptor CCR5 in tick-borne encephalitis

BackgroundChemokine receptor 5 (CCR5) is hypothesized to drive the lymphocyte migration to central nervous system in flavivirus encephalitis, and the non-functional CCR5Δ32 genetic variant was identified as a risk factor of a West Nile virus infection and of tick-borne encephalitis (TBE). We have attempted to investigate how CCR5 expression corresponds to the clinical course and severity of TBE.MethodsWe have repeatedly studied CCR5 expression in 76 patients during encephalitic and convalescent TBE phase, analyzing its association with clinical features, cerebrospinal fluid (csf) pleocytosis, and concentrations of CCR5 ligands (chemokines CCL3, CCL4, and CCL5) and CCR5 genotype. Fifteen patients with neuroborreliosis, 7 with aseptic meningitis, 17 in whom meningitis/encephalitis had been excluded, and 18 healthy blood donors were studied as controls. Expression of CCR5 was measured cytometrically in blood and csf-activated Th lymphocytes (CD3+CD4+CD45RO+). Concentrations of chemokines in serum and csf were measured immunoenzymatically, and CCR5Δ32 was detected with sequence-specific primers. Data were analyzed with non-parametric tests, and p < 0.05 was considered significant.ResultsThe blood expression of CCR5 did neither differ between the groups nor change in the course of TBE. The CCR5 expression in the inflammatory csf was several-fold increased in comparison with blood but lower in TBE than in neuroborreliosis. The csf concentration of CCL5 was increased in TBE, the highest in the most severe presentation (meningoencephalomyelitis) and correlated with pleocytosis. The CCR5Δ32/wt genotype present in 7 TBE patients was associated with a decreased CCR5 expression, but enrichment of csf Th population in CCR5-positive cells and the intrathecal inflammatory response were preserved, without a compensatory increase of CCL5 expression.ConclusionsWe infer CCR5 and CCL5 participate in the response to TBE virus, as well as to other neurotropic pathogens. The intrathecal response to TBE is not hampered in the bearers of a single copy of CCR5Δ32 allele, suggesting that the association of CCR5Δ32 with TBE may be mediated in the periphery at the earlier stage of the infection. Otherwise, a variability of the CCR5 expression in the peripheral blood lymphocytes seems not to be associated with a variable susceptibility to TBE.

Platelet indices in SGA newborns

PURPOSE The current study objective was to compare blood platelet indices in full-term small-for-gestational-age newborns (SGA) and full-term appropriate-for-gestational-age newborns (AGA). MATERIALS/METHODS We introduced to our study 61 SGA newborns (31 females and 30 males) and 70 eutrophic infants (32 females and 38 males). The SGA newborns were divided into two groups: those weighing less than the 5th centile: 35 infants (16 females and 19 males) and those between the 5th and 10th centiles: 26 infants (15 females and 11 males). Platelet indices were estimated in blood samples collected from the umbilical artery. RESULTS SGA demonstrated a decreased count of blood platelets (238×103/μ) as compared with AGA (286×103/μL), p=0.0001. Platelet hematocrit (PTC) also showed differences in both groups (SGA=0.19% vs. AGA=0.22%; p=0.0005). Mean platelet volume (MPV) was higher in SGA (8.25fl) as compared with AGA (7.84fl); p=0.008. Large platelet count (LPLT) was higher in AGA 6.26% vs. SGA=4.75%; p=0.01. Platelet distribution width (PDW) was found to be nearly the same (SGA=47%, AGA=46%). PDW was higher in SGA newborns < 5th centile (43%) as compared with SGA infants between the 5th and 10th centiles (52%); p=0.008. CONCLUSIONS A decreased blood platelet count, platelet hematocrit and large metabolically active platelet count, which in addition to reduced synthesis and excessive consumption of coagulation factors in states of hiperclotting is characteristic of IUGR, enhances the possibility of bleeding complications and increases the risk of infections. From a clinical point of view, it is important to take into consideration the degree of intrauterine hypotrophy during the evaluation of hemostatic disorders.

Potential role of β1 integrin and collagen biosynthesis in estrogen-dependent reduction of apoptosis in tamoxifen-treated breast cancer cells

It was found that 10 µM tamoxifen induced apoptosis and a significant (approximately 50%) depletion of β1 integrin levels in human breast cancer cells. Estradiol-treated MCF-7 cells exhibited exceptional viability and adherence, high levels of β1 integrin and increased (by 100%) collagen biosynthesis. Pretreatment of MCF-7 cells with 1 nM estradiol prevented tamoxifen-induced cell death, loss of cell adherence and decrease in β1 integrin level. Tamoxifen and estradiol had an opposite effect on the β1 integrin level and adherence in breast cancer cells, suggesting that the decrease in the β1 integrin level may be an early event during tamoxifen-induced apoptosis in breast cancer cells.

Platelet activation in acute pancreatitis.

Objectives The aim of this study was to assess the functional state of platelets in patients with mild acute pancreatitis and severe acute pancreatitis (S-AP). Methods The number of platelets and their morphological parameters were measured with Advia 2120. &bgr;-Thromboglobulin and platelet factor 4 concentrations were determined by enzyme-linked immunosorbent assay method. To evaluate the expression of platelet glycoproteins, flow cytometry method was used. Results At the time of admission, a multiparameter evaluation of the platelets’ function in AP patients showed enhanced platelet activation, which was reflected by an increase in the number of large platelets, concentration of degranulation markers (platelet factor 4 and &bgr;-thromboglobulin), expression of glycoprotein (Gp) IIb/IIIa, and decreased mean platelet component. Only in S-AP patients at day 1 a decreased number of platelets and high expression of P-selectin and GpIa were observed, which may suggest their prognostic value. At day 30, the procoagulation state was still present in S-AP patients, because of increased platelets and number of large platelets as well as high GpIIb/IIIa expression. Conclusions These results may indicate an important role of platelet activation in the pathogenesis of acute pancreatitis and the development of complications in S-AP.

Utility of laboratory tests in B-CLL patients in different clinical stages

The study objective was to analyse the utility of laboratory tests performed in 30 patients with B-cell chronic lymphocytic leukaemia (B-CLL) at different clinical stages. Laboratory tests included automated and microscopic assessment of peripheral blood and bone marrow counts as well as evaluation of leukaemic cells. Apart from the diagnostic and prognostic value of laboratory abnormalities such as clonal lymphocytosis with CD5+CD19+CD23+ phenotype, reduced erythrocyte parameters, thrombocytopenia or bone marrow infiltration by the neoplastic clone as well as low percentage of Gumprecht’s shadows, low apoptotic activity of peripheral blood lymphocytes, and increased percentage of CD38− and ZAP-70 ± cells markedly correlate with the stage of disease progression. These results seem to confirm the diagnostic and prognostic significance of these parameters determined in routine laboratory tests in B-CLL patients.

Expression of Fas receptor on human T lymphocytes under stimulation with Borrelia burgdorferi sensu lato – preliminary results

PURPOSE Apoptosis of activated T lymphocytes is essential to immunoregulation and its abnormalities have been observed in immune system disorders and persistent infections. To asses Borrelia burgdorferi influence on the susceptibility of T lymphocytes to apoptosis, we have measured expression of the Fas death receptor on these cells after incubation with live B. burgdorferi. MATERIAL AND METHODS Peripheral blood mononuclear cells from 23 LD patients (18 with Lyme arthritis, 5 with neuroborreliosis) and 13 healthy controls (C) were incubated for 48 hours with and without live B. burgdorferi spirochetes: B. afzelii, B. garinii or B. burgdorferi sensu stricto. After incubation, Fas expression on CD3+ cells was measured cytometrically with FITC-labeled monoclonal antibody. RESULTS Median fraction of Fas-expressing T lymphocytes increased under incubation with B. burgdorferi, with more cells expressing Fas after incubation with B. burgdorferi sensu stricto than with B. garinii. There was a tendency for a higher expression of Fas on T lymphocytes from LD patients then from controls, both in unstimulated and B. burgdorferi-stimulated cultures, but it did not reach a level of statistical significance. CONCLUSIONS B. burgdorferi seems to increase Fas expression on CD3+ T lymphocytes, which may render these cells more susceptible to apoptosis. This effect is stronger for B. burgdorferi s.s. than for B. garinii genospecies.

Plasma levels of MMP-7 and TIMP-1 in laboratory diagnostics and differentiation of selected histological types of epithelial ovarian cancers

BackgroundMMP-7 and TIMP-1 may play a role in the pathogenesis of cancer disease. In this study we investigated plasma levels of selected metalloproteinase and its tissue inhibitor in comparison to plasma levels of the commonly accepted tumor markers (CA 125 and HE4) in selected histological types of epithelial ovarian cancer patients as compared to control groups: patients with a benign ovarian tumor and healthy subjects. Plasma levels of MMP-7 and TIMP-1 were determined using ELISA, CA 125 and HE4 – by CMIA methods.ResultsPlasma levels of all biomarkers studied were significantly higher in ovarian cancer patients as compared to both control groups. MMP-7 demonstrated comparable to HE4 or CA125 values of diagnostic sensitivity (SE: 61%; 68%; 58%, respectively), specificity (SP: 95%; 95%; 98%, respectively), positive (PPV: 93%; 96%; 98%, respectively) and negative predictive values (NPV: 61%; 66%; 60%, respectively) in the groups tested. The combined use of the aforementioned biomarkers resulted in a further increase in diagnostic criteria and AUC, especially in the early stages of the disease.ConclusionsThese findings suggest the usefulness of combining MMP-7 with CA 125 and HE4 in the diagnosis of epithelial ovarian cancer as a new tumor marker panel.