Psoriasis and metabolic syndrome in children: current data.

BACKGROUND: The prevalence of cardiovascular and metabolic disorders in paediatric patients with psoriasis is not well established. AIM: To conduct a meta-analysis of previously published studies dealing with the occurrence of metabolic disorders in children with psoriasis. METHODS: Data from 7 studies with a total of 965 children with psoriasis were analysed using a random effects model. RESULTS: Prevalence of metabolic syndrome (MetS) was significantly higher in patients with psoriasis than in healthy controls (HCs). In most studies, significantly decreased levels of high-density lipoprotein (HDL) cholesterol were found in children with psoriasis. Mean level of HDL cholesterol in patients with psoriasis was 2.05 mg/dL lower than in HCs. Patients with psoriasis and HCs did not differ significantly in their mean triglyceride levels, although the difference was at a threshold of statistical significance. Mean level of fasting glucose in children with psoriasis was 5.75 mg/dL higher than in HCs (P < 0.01). The two groups did not differ significantly in mean waist circumference or in systolic and diastolic arterial pressures. CONCLUSIONS: Decreased levels of HDL cholesterol and increased concentrations of fasting glucose may represent very early stages of MetS in children with psoriasis. However, a large population-based study is needed to establish the relationship between psoriasis and MetS in children, including the environmental, genetic and immunological factors leading to their co-occurrence.

TL1A as a Potential Local Inducer of IL17A Expression in Colon Mucosa of Inflammatory Bowel Disease Patients.

Interaction between TL1A and death receptor 3 (DR3) co-stimulates T cells, induces production of several pro-inflammatory cytokines and has been linked to pathogenesis of inflammatory bowel disease (IBD). This study aimed to establish a link between expression of TL1A and selected TL1A-induced pro-inflammatory cytokines involved in IBD pathogenesis (IL-4, IL-13, IL-17A and IFN-γ) and to investigate a connection between serum concentration of TL1A in patients with IBD and activation of peripheral blood T cells. Elevated levels of IL-4 (2.91-fold) and IL-13 (4.05-fold) mRNA were detected in the inflamed colon mucosa of patients with ulcerative colitis (UC), IFN-γ mRNA was upregulated (3.23-fold) in the inflamed colon mucosa of patients with Crohn's disease (CD), whereas upregulation of IL-17A and TL1A mRNA was present in the inflamed colon mucosa of patients with both CD and UC (IL-17A: 4.48-fold and 2.74-fold, TL1A: 3.19-fold and 3.22-fold, respectively) vs. control subjects. We did not detect any changes in DR3 mRNA expression in the investigated groups of patients. TL1A mRNA level in colon mucosa of patients with IBD correlated only with the level of IL-17A mRNA but no other investigated cytokines. In colon mucosa, expression of TL1A and DR3 was localized to enterocytes and lamina propria mononuclear cells. We did not find any correlation between serum concentrations of TL1A and IL-17A or changes of CD4(+) or CD8(+) lymphocytes phenotype in patients with IBD. Therefore, our data indicate that TL1A may contribute to pathogenesis of IBD via local but not systemic induction of IL-17A but not IL-4, IL-13 or IFN-γ.

mtDNA(4977) deletion is not a common feature in patients with premature ovarian failure and primary infertility.

The aim of the current study was to investigate the incidence of mtDNA(4977) deletion in peripheral blood leukocytes of patients diagnosed with premature ovarian failure (POF) and primary infertility. The study group consisted of 17 patients with POF, 32 women with primary infertility, and 31 fertile women with the prevalence of the mtDNA(4977) deletion using the reverse transcription-polymerase chain reaction (RT-PCR) based technology. None of the patients affected by POF revealed mtDNA(4977) deletion. This deletion was detected only in one 26-year-old infertile patient. No significant difference in relation to mtDNA(4977) deletion was reported between the groups investigated (p > 0.05). In conclusion, mtDNA(4977) deletion is not a common finding in peripheral blood leukocytes of women affected by POF and primary infertility. The occurrence of mtDNA(4977) deletion in women between 20 and 39 years of age may not increase with increasing patients' age, independently of their fertility status.

Abnormal Expression of BTLA and CTLA-4 Immune Checkpoint Molecules in Chronic Lymphocytic Leukemia Patients.

Chronic lymphocytic leukemia (CLL) is characterized by the peripheral accumulation of neoplastic B cells and is frequently complicated by the systemic immunosuppression associated with an impairment in B and T lymphocyte activation. We hypothesized that the expression of immune checkpoint suppressors B and T lymphocyte attenuator (BTLA) and cytotoxic T lymphocyte antigen (CTLA-4) is disturbed in both lymphocyte subpopulations in CLL. The expression of CTLA-4 and BTLA mRNA was determined by real-time PCR, while CTLA-4 protein expression (surface or intracellular) was estimated in BTLA+ lymphocytes by flow cytometry. In CLL patients, we observed a higher gene transcript level of BTLA and CTLA-4 than in healthy individuals in both freshly isolated and PMA stimulated B and T cells. Remarkably, lower amounts of both inhibitory proteins were found in peripheral blood (PB) CLL B cells, whereas normal BTLA and elevated CTLA-4 were found in T cells. Consistently, there was a prevalence of CTLA-4+ cells within circulating BTLA+ T cells cells of patients confronting PB healthy cells. After in vitro stimulation, the only change found in CLL patients was a decrease in BTLA expression in B and T lymphocytes. In contrast, healthy lymphocytes responded more vigorously as regards the BTLA and CTLA expression with substantially higher frequency of CD69+ cells under the stimulating condition compared to corresponding cells from the CLL group. Our results indicate that CLL development is associated with the affected expression of BTLA and CTLA-4 checkpoint receptors in PB and its impaired expression might be associated with lowering of the threshold for B cell activation and proliferation, while upregulated CTLA-4 expression in CLL peripheral BTLA+ T cells may contribute to suppressed T cell effector functions. This hypothesis needs to be validated in future studies, which would allow us to explain how the increased or decreased expression of these molecules affects the cell function.

The significance of soluble HLA-G plasma levels as well as messenger HLA-G for B-cell chronic lymphocytic leukemia (B-CLL).

The immunosuppression accompanies B-cell chronic lymphocytic leukemia (B-CLL) but might be also responsible for disease progression by enabling CLL cells to escape from the immunosurveillance. Some particles involved in the regulation of an immune system might represent prognostic value for B-CLL. Recently we found no correlation between HLA-G on messenger and protein level, suggesting that HLA-G is released in soluble form. To confront this hypothesis we characterized soluble HLA-G (sHLA-G) by the prognostic factors in the first cohort of 34 CLL patients. No correlation was observed between sHLA-G levels in ZAP-70(+) and ZAP-70(-) CLL as well as in CD38(+) CLL and CD38(-) CLL patients. Next, we wondered whether gene expression of HLA-G, which represent the whole HLA-G pool in the cell, posses prognostic value for CLL. In the second cohort of 41 CLL patients we assessed messenger levels of HLA-G by the strongest prognostic factors in CLL including cytogenetics, IgVH mutational status, ZAP-70 as well as CD38. No changes of HLA-G expression levels were found in different CLL groups characterized by IgVH gene mutational status, ZAP-70 as well as CD38. We observed no differences in expression of HLA-G in various cytogenetic groups of CLL including del17p, del13q, del11q, +8q, +3q, del14q and del6q when compared to those with normal karyotype or with 12+. Both, mRNA expression of HLA-G and levels of its soluble form in plasma bring no additional prognostic value for B-CLL patients.

Expression of RHAMM/CD168 and other tumor-associated antigens in patients with B-cell chronic lymphocytic leukemia.

Antigen targeted immunotherapies might represent a novel treatment for B-cell chronic lymphocytic leukemia (B-CLL). We screened the mRNA expression of tumor-associated antigens (TAAs) from the literature (fibromodulin, survivin, OFA-iLRP, BAGE, G250, MAGE1, PRAME, proteinase, syntaxin, hTERT, WT-1) and TAAs defined previously by serological analysis of cDNA expression libraries from leukemic cells (PINCH, HSJ2, MAZ, MPP11, RHAMM/CD168, NY-Ren60). Peripheral blood mononuclear cells from 43 B-CLL patients and 20 healthy volunteers (HVs) were examined by conventional and quantitative RT-PCR. mRNA of RHAMM/CD168, fibromodulin, syntaxin and NY-Ren60 was expressed in 55-90%, and mRNA of HSJ2, MAZ and OFAiLRP was expressed in 90-100% of the patients. No expression of WT-1, hTERT, BAGE, G250, MAGE1 or survivin was observed. Low (2-20%) expression frequencies of MPP11, PINCH, PRAME and proteinase were detected. RHAMM/CD168, fibromodulin, PRAME and MPP11 showed expression in B-CLL patients, but not in HVs. Because of the exquisite tissue expression of RHAMM/CD168 and its high expression frequency in CLL patients, mixed lymphocyte peptide culture (MLPC), enzyme-linked immunosorbent spot (ELISPOT) and flow cytometry were performed for antigen specific T-cells. In MLPC, RHAMM specific responses by CD8+HLA-A2/R3tetramer+CCR7-CD45RAhigh effector T-cells were detected. RHAMM/CD168 might be a possible target for future immunotherapies in both ZAP-70(+) and ZAP-70(-) B-CLL patients.

Allogeneic dendritic cells pulsed with tumor lysates or apoptotic bodies as immunotherapy for patients with early-stage B-cell chronic lymphocytic leukemia.

Recently, immunotherapies with allogeneic dendritic cells (DCs) pulsed with tumor antigens to generate specific T-cell responses have been tested in clinical trials for patients with solid tumors. This is the first report on a clinical vaccination study with DCs for patients with B-cell chronic lymphocytic leukemia (B-CLL). The potential of allogeneic DCs pulsed ex vivo with tumor cell lysates or apoptotic bodies to stimulate antitumor immunity in patients with B-CLL in early stages was evaluated. Monocyte-derived DCs were obtained from unrelated healthy donors. Nine patients (clinical stage 0 and 1 according to Rai) were vaccinated five times with a mean number of 32 x 10(6) stimulated DCs administered intradermally once every 2-3 weeks. No signs of autoimmunity were detected, and only mild local skin reactions were noted. During the treatment period, we observed a decrease of peripheral blood leukocytes and CD19+/CD5+ leukemic cells. In one patient, a significant increase of specific cytotoxic T lymphocytes against RHAMM/CD168, a recently characterized leukemia-associated antigen, could be detected after DC vaccination. Taken together, the study demonstrated that DC vaccination in CLL patients is feasible and safe. Immunological and to some extent hematological responses could be noted, justifying further investigation on this immuno-therapeutical approach.

The influence of thalidomide therapy on cytokine secretion, immunophenotype, BCL-2 expression and microvessel density in patients with resistant or relapsed multiple myeloma.

Thalidomide (THAL) is currently used as a novel drug in patients with chemotherapy resistant or relapsed multiple myeloma. THAL antitumor activity seems to be very complex, however the precise mechanisms of its action are still not fully understood. The aim of this study was to assess some of possible mechanisms of THAL action both in in vivo analysis of immune cells phenotype and in in vitro cultures with THAL. The study involved 30 patients with relapsed or chemotherapy refractory multiple myeloma who were qualified to THAL treatment. We assessed immunophenotype of malignant plasma cells and T lymphocytes in both peripheral blood (PB) and bone marrow (BM) samples taken before and after 4 and 8 weeks of THAL treatment. Before therapy cytokine secretion (VEGF, HGF, bFGF, TNF, IL-6 an sIL-6R) and Bcl-2 expression in PB and BM cell cultures with THAL were analyzed. We used flow cytometry technique and ELISA method. The clinical response to therapy was assessed after 4 and 8 weeks of treatment. We also investigated microvessel density (MVD) in bone marrow samples before the THAL treatment and after 6 months of therapy in the group of responding patients. In cell cultures with THAL we detected statistically significant lowering of analyzed cytokines concentration and the decrease in Bcl-2 expression by malignant plasma cells in BM and CD8(+) T lymphocytes in BM and PB. In the group of patients responding to therapy we observed the decrease in the number of myeloma cells and significant increase of CD4(+) and CD8(+) cells in both PB and BM samples. There was statistically significant increase of CD3(+)/CD69(+) cells in the course of therapy, while the percentage of CD3+/HLA-DR(+) cells was significantly lower after 8 weeks of therapy. We also detected lowering of MVD after THAL therapy in responders group. The obtained results demonstrate that THAL efficacy in MM is multidirected and included such mechanisms like down-regulation of proangiogenic cytokines, that could lead to lowering of MVD, induction of apoptosis and influence on malignant cells and T lymphocytes immunophenotype.

Apoptosis of HeLa cell lines incubated with retinol.

PURPOSE: The aim of the study was to evaluate a soluble form of APO-1/Fas antigen in supernatants from HeLa cell line culture after 24 and 72 h of incubation with selected retinoic acid concentrations. MATERIALS AND METHODS: HPV18-positive cell lines were cultivated with All-trans-retinoic acid (ATRA) at concentrations of 1 x 10(-6) and 1 x 10(-9) M. The cultures were incubated for 24 and 72 h. A control culture with 3 microl of DMSO was incubated under identical conditions. The concentrations of soluble APO-1/Fas antigen in cell culture supernatants were estimated using an ELISA method. RESULTS: The culture with 72-h incubation with retinoic acid proved to be toxic to cells and was excluded from the analysis. The results obtained showed a significantly lower concentrations of soluble APO-1/Fas antigen in supernatants from cell lines incubated with retinol for 24 h than in the controls. CONCLUSIONS: The higher concentrations of soluble APO-1/Fas antigen in supernatants from the HeLa cell line without retinol may constitute a protective mechanism of the cells infected with the virus before undergoing Fas/FasL-dependent apoptosis. Lower concentrations of sAPO-1/Fas antigen in the supernatant from HeLa cell culture incubated with retinol may suggest that mechanisms protecting infected cells against Fas/FasL-mediated apoptosis become defective under the influence of retinol. Our studies confirm that Vitamin A and its analogues inhibit the proliferation of cells associated with HPV infection and suggest promising effects of retinoid therapy in inhibiting the progression of early cervical lesions to cancer.

The degree of lymphocytic mitochondrial transmembrane potential and blood magnesium concentrations during coronary artery bypass grafting.

UNLABELLED: Magnesium (Mg) plays an important role in lymphocyte function. Low blood concentration of Mg may result in intralymphocyte imbalance and in turn may be associated with intensified apoptosis of peripheral blood lymphocytes. Due to its multistage character; extracorporeal circulation (ECC) may augment Mg disorders adding to the above mentioned pathology. The aim of this study was to assess the correlation between lymphocyte apoptosis and Mg concentration in the blood during the course of coronary artery bypass grafting (CABG) and in the early postoperative period. METHOD: Twenty male patients undergoing CABG with ECC under general anaesthesia were included in the study. For detection of apoptotic lymphocytes in the circulation, inner mitochondrial transmembrane potential (deltapsim) was measured with the use of chloromethyl-X-rosamine (CMXRos) and flow cytometry. Spectrophotometry was used for Mg blood concentration measurements. Peripheral blood samples were obtained in seven stages: 1) just before anaesthesia, 2) 2 hours after the beginning of surgery, 3) immediately after surgery, 4) 12 hours after the beginning of surgery, 5) 24 hours after the beginning of surgery, 6) 36 hours after the beginning of surgery, 7) 54 hours after the beginning of surgery. RESULTS: The statistically significant increases of lymphocyte apoptosis were noted in stages from 2 to 7. Blood Mg concentrations decreased in stages 2 and 3. There was negative correlation between Mg blood concentration in stages 2 and 3 and the intensity of lymphocyte apoptosis in the stage 5. CONCLUSIONS: 1) CABG with extracorporeal circulation was associated with a decrease of magnesium concentration in the blood and an increase of lymphocyte apoptosis intensity. 2) The decrease of magnesium blood concentration may increase the degree of lymphocyte apoptosis. 3) Lymphocyte apoptosis after extracorporeal circulation has a two-phase course.

The effect of 2-h infusion of 2-chlorodeoxyadenosine (cladribine) with prednisone in previously untreated B-cell chronic lymphocytic leukaemia.

2-Chlorodeoxyadenosine (2-CdA) is a new antimetabolite chemotherapeutic agent active in indolent lymphoid malignancies. In this retrospective study, 69 previously untreated patients with B-cell chronic lymphocytic leukaemia (B-CLL) were treated with 2-CdA administered at a dose of 0.12 mg/kg daily in 2-h intravenous infusion for 5 consecutive days. 45 patients also received prednisone 30 mg/m2 orally each day for 5 days starting with 2-CdA courses. Patients were given 2-6 courses (mean 4.6) of 2-CdA repeated usually at monthly intervals. If a complete response was achieved, no further 2-CdA courses were administered. Guidelines for response were those developed by the NCI Sponsored Working Group. Complete response (CR) was achieved in 26 (38%) and partial response (PR) in 27 (39%) cases, giving an overall response rate of 77%. 16 patients (23%) did not respond to 2-CdA. In the subgroup of 45 patients receiving 2-CdA with prednisone, CR was obtained in 15 (33%) and PR in 20 (44%) patients giving an overall response rate of 78%. CR was achieved in 11 (46%) out of 24 patients treated only with 2-CdA and in 7 cases (29%) PR was observed, giving an objective response rate of 75%. The differences between both subgroups were not statistically significant. However, we observed a relationship between the response and the number of courses of 2-CdA given in patients receiving and those not receiving prednisone. In the subgroup receiving 2-CdA with prednisone, an earlier response to 2-CdA was observed. In this group a response was achieved in 9 (20%) patients after two courses of 2-CdA and in 18 (40%) after four courses. In the subgroup receiving only 2-CdA, 17 (71%) responses were obtained after six cycles.

Pre-eclampsia affects the immunophenotype of neonates.

OBJECTIVE: We tried to estimate whether immunological changes are present in neonates born to mothers who had been suffering from pre-eclampsia. STUDY DESIGN: Eighteen neonates born to mothers with severe pre-eclampsia (between 35 and 40 weeks of gestation) and 20 full-term healthy newborns (between 38 and 40 weeks of gestation) were included in the study. The lymphocytes were isolated from umbilical cord blood. The specific lymphocyte antigens were determined using direct staining with monoclonal antibodies and analysed by flow-cytometry. RESULTS: We observed that neonates born to pre-eclamptic mothers had decreased percentage of T CD 3(+), CD 4(+) and T CD 8(+)28(+) (cytotoxic) lymphocytes and increased percentage CD 3(-)16/56(+) cells and CD 8(+)28(-) (suppressor) lymphocytes in comparison with newborns of healthy women. Furthermore, we found decreased CD 4: CD 8 lymphocyte ratio in the study group in comparison with the control group. We also observed that the percentage of CD 19(+)5(+), CD 4(+)8(+), CD 19(+)40(+) and CD 3(+)40L(+) lymphocytes did not differ in both studied groups. The percentage of CD 4(+)45RO(+), CD 8(+)45RO(+) memory cells was higher in neonates born to pre-eclamptic mothers when compared to controls. Moreover, the expression of CD 25 molecule was higher on T CD 8(+) and B CD 19(+) lymphocytes of neonates of pre-eclamptic mothers. CONCLUSION: The alterations in the immunological parameters of neonates born to pre-eclamptic mothers can be associated with the maternal disease.

CD1c(+) immature myeloid dendritic cells are predominant in cord blood of healthy neonates.

It has been suggested lately that some types of antigen presenting cells-myeloid dendritic (DC-1) cells can differentiate the immune response towards Th1 type immunity, whereas lymphoid cells (DC-2) can stimulate Th2 type immunity. It has been observed that neonates are deficient in Th1 response. The purpose of our study was to estimate the proportions of immature myeloid (CD1c(+)) and lymphoid (BDCA-2(+), BDCA-4(+)) dendritic cells and the CD1c(+):BDCA-2(+) cell ratio in cord blood of healthy neonates in comparison with dendritic cells of healthy adults. Thirty healthy neonates born from normal pregnancies and 30 healthy adults were included in the study. The dendritic cells were isolated from cord and peripheral blood, stained with anti-CD1c, anti-BDCA-2, anti-BDCA-4, anti-CD123 and anti-CD19 monoclonal antibodies and estimated using flow cytometry. The percentage of CD1c(+) dendritic cells in cord blood of healthy newborns did not differ significantly when compared to those in peripheral blood of healthy adults. The percentages of cord blood BDCA-2(+) and BDCA-4(+) dendritic cells of neonates were significantly lower when compared to lymphoid dendritic cells in peripheral blood of adults. The CD1c(+):BDCA-2(+) ratio was significantly higher in cord blood of neonates in comparison with CD1c(+):BDCA-2(+) ratio in adult's blood. Myeloid and lymphoid dendritic cells may be involved in the immune regulation during fetal development. Immature myeloid dendritic cells are predominant in cord blood of healthy neonates. Immature lymphoid dendritic cells are not the major population of dendritic cells in cord blood.

The expression and concentrations of Fas/APO-1 (CD95) antigen in patients with severe pre-eclampsia.

PROBLEM: Apoptosis has been proposed as a mechanism for maintaining homeostasis in the immune system. Activated lymphocytes are removed by a Fas/FasL-mediated programmed cell death process called activation induced cell death (AICD). The aim of the study was to investigate surface Fas antigen (APO-1, CD95) expression on T lymphocytes and NK cells and also soluble Fas antigen concentrations in pre-eclamptic patients and healthy pregnant women. MATERIALS AND METHODS: Sixteen pre-eclamptic patients and 18 healthy pregnant women were studied. Peripheral blood lymphocytes were isolated, labeled by direct staining with anti-CD95 monoclonal antibodies and analyzed using the flow cytometric method. Furthermore, the concentrations of soluble CD95 molecule in serum of patients with severe pre-eclampsia and women with uncomplicated pregnancy were measured using ELISA method. RESULTS: We found that Fas antigen expression and fluorescence intensity on T CD8+ lymphocytes were higher in patients with severe pre-eclampsia in comparison with healthy pregnant women (P<0.05). Furthermore, the concentrations of soluble CD95 molecule were higher in the group of pre-eclamptic patients when compared to controls (P<0.001). CONCLUSIONS: These findings suggest that T CD8+ lymphocytes in patients with severe pre-eclampsia can be activated. Moreover, higher concentrations of soluble CD95 antigen can suggest altered possibilities to undergo Fas/FasL-mediated activation induced cell death process of lymphocytes in severe pre-eclampsia.

Blood myeloid and lymphoid dendritic cells are stable during the menstrual cycle but deficient during mid-gestation.

The aim of this study was to estimate the populations of peripheral blood myeloid and lymphoid dendritic cells (CD1c(+), BDCA-2(+), BDCA-4(+)) and the CD1c(+):BDCA-2(+) ratio in phases of the ovarian cycle and in normal pregnant patients. 18 non-pregnant women and 17 normal pregnant women were included. Dendritic cells were isolated from peripheral blood, stained with monoclonal antibodies (mAbs) against blood dendritic cell antigens (anti-BDCA-1, BDCA-2, BDCA-4) and estimated using flow cytometry. CD1c(+), BDCA-2(+) and BDCA-4(+) dendritic cells were present in the follicular and luteal phases of the ovarian cycle and in all trimesters of normal pregnancy. The percentages of CD1c(+) dendritic cells did not differ between the follicular and luteal phases of the ovarian cycle. The percentage of BDCA-2(+) dendritic cells was lower in the luteal phase of the ovarian cycle compared with the follicular phase, but the differences were not statistically significant. The CD1c(+):BDCA-2(+) cell ratio was significantly lower in the luteal phase compared with the follicular phase of the ovarian cycle. The numbers of dendritic cells were significantly lower in the second trimester when compared with the first and third trimesters of normal pregnancy. Furthermore, in the second trimester, the CD1c(+):BDCA-2(+) ratio was higher than in the other trimesters of normal pregnancy. All populations of dendritic cells and the CD1c(+):BDCA-2(+) ratio did not differ in the first and third trimesters of physiological pregnancy. Our results suggest that myeloid and lymphoid dendritic cells are not affected by steroid hormones during the menstrual cycle. The deficiency of peripheral blood dendritic cells observed during the second trimester of normal pregnancy can be associated with their migration to the uterus during the second physiological invasion by cytotrophoblast.

Influence of low dose rIL-2 treatment on endogenous cytokine production, expression of surface IL-2R and the level of soluble IL-2R in patients with minimal residual disease.

This study was designed to investigate the immunomodulatory effect of low-dose IL-2 therapy (100 microg/day for 3 weeks) on interferon (IFN), tumor necrosis factor (TNF) production in vivo and in vitro and on the expression of IL-2Ralpha/beta and soluble form of IL-2Ralpha. Patients enrolled in the study suffered from multiple myeloma (MM), Hodgkin's disease (HD) and non-Hodgkin's lymphoma (NHL) All of them were in remission after chemotherapy or radiotherapy. Our results indicated that IL-2 given subcutaneously at a low dose of 100 microg/day for 3 weeks induced IFN-gamma and TNF-alpha in plasma (measured 24 hrs after the last dose of IL-2) and affected the ability of blood leukocytes to produce cytokines. Production of IFN-gamma induced in vitro with PHA was enhanced, but TNF-alpha production induced by lipopolysaccharide (LPS) and virus (Newcastle Disease Virus) was depressed. The expression of both: surface IL-2R, especially beta subunit on total population of lymphocytes and NK cells, and soluble form of IL-2R, of chain were significantly enhanced after low-dose IL-2 therapy. Low dose IL-2 therapy was well tolerated by all patients, and side effects not exceeding II grade of toxicity according to WHO scale were observed. Five patients with MM relapsed 3-10 month after cessation of IL-2 therapy, but all patients with Hodgkin's and non-Hodgkin's lymphomas are still in remission (20 months of observation).

Interferon production by peripheral blood leukocytes of patients with acute myeloid leukemia.

The ability of peripheral blood leukocytes (PBL) of 13 selected patients with acute myeloid leukemia (AML), and of 10 healthy subjects to produce interferon (IFN) spontaneously and after in vitro induction was tested. Spontaneous IFN production was detected in supernatants of PBL cultures of healthy subjects but was not present in cultures of leukemic cells (leukocytes density 1 x 10(6) ml). After induction with Newcastle disease virus (NDV) PBL cultures from 5 patients with AML exhibited a low IFN response, while in others IFN titers were similar to controls. The IFN titers induced with lipopolysaccharide (LPS) were lower in leukemic cells than those detected in control cells. The absence or low IFN levels after induction with phytohemagglutinin (PHA) correlated with very small numbers of normal lymphocytes in blood of leukemic patients.

Vascular endothelial growth factor and basic fibroblast growth factor in patients with squamous cell oesophageal cancer.

It is suggested that vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (bFGF) play an important role in tumor-induced angiogenesis. The purpose of this study was to estimate the correlation between VEGF and bFGF levels and tumor pathological status according to pTNM classification in patients with squamous cell oesophageal cancer. A group of 25 healthy controls and 32 consecutive patients with oesophageal cancer were included in this study. Serum VEGF and bFGF levels were determined by enzyme-linked immunosorbent assay (Quantikine R&D Systems). Serum VEGF and bFGF levels were significantly elevated in the patient groups (VEGF: 146.0 pg/ml, 79.0-386.3 pg/ml vs. 38.0 pg/ml, 6.5-135.1 pg/ml, p<0.005, and bFGF: 5.2 pg/ml, 1.2-10.6 pg/ml vs. 2.06 pg/ml, 0.07-4.0 pg/ml, p<0.02 Fisher test). The highest correlation between serum VEGF and bFGF levels were found in patients with advanced cancers, especially with: T4, N1, and M1 factors. The VEGF and bFGF levels were significantly higher in patients with pT4 (p<0.01). Patients with N1 lymph node invasion, compared with N0 factor, have higher levels of angiogenetic factors (p<0.04). Also in patients with advanced cancers with liver metastases the serum levels VEGF and bFGF were significantly higher (M1 vs. M0, VEGF p<0.001 and bFGF p<0.05). Consecutive monitoring of VEGF and bFGF serum levels may be a useful prognostic marker for patients with squamous cell oesophageal cancer.

T helper 1- and T helper 2-type cytokine imbalance in pregnant women with pre-eclampsia.

OBJECTIVES: The purpose of our study was to investigate T helper 1/T helper 2 balance in pregnant women with pre-eclampsia. STUDY DESIGN: 18 patients with pre-eclampsia and 20 healthy pregnant women were included in the study. Peripheral blood mononuclear cells (PBMC) were stimulated with phytohaemagglutinin (PHA) for 48 h. Cytokine: interleukin-2 (11-2), interferon-gamma (IFN-gamma) and interleukin-10 (I1-10) concentrations in culture supernatants were determined using the ELISA method. Statistical analysis was performed using a standard non-parametric Mann-Whitney U-test. RESULTS: We found that in pre-eclamptic patients PHA-stimulated 11-2 and IFN-y production was significantly higher (P<0.001) and I1-10 production significantly lower (P<0.005) in comparison with the control group. CONCLUSION: These results could suggest that there is Th1/Th2 imbalance in pre-eclamptic patients with predominant Th1-type immunity.

The concentrations of osteocalcin and degradation products of type I collagen in pregnant women with pre-eclampsia.

OBJECTIVES: To investigate the concentrations of osteocalcin and collagen type I C-terminal telopeptides in pregnant women with pre-eclampsia. STUDY DESIGN: 26 patients with severe pre-eclampsia and 24 healthy pregnant women were included in the study. Serum concentrations of osteocalcin and C-telopeptides--degradation products of type I collagen were determined using the ELISA method. Statistical analysis was performed using Mann-Whitney U-test. RESULTS: In pre-eclamptic patients, the concentrations of osteocalcin and degradation products of collagen type I were significantly higher (P<0.005) when compared to healthy pregnant women. CONCLUSION: These results could suggest that there are alterations in bone metabolism in pregnant women with pre-eclampsia.