Sub-optimal primary surgery leads to unfavorable immunological changes in ovarian cancer patients.

Primary cytoreduction, followed by chemotherapy, is a standard treatment of patients with epithelial ovarian cancer (EOC). However, the effectiveness of this treatment depend on various elements e.g. type of operation. It is accepted that optimal surgery correlates with longer survival of patients. The other element, an efficiency of immune system after surgical intervention although important is less elucidated. The aim of this study was to establish the impact of optimal and sub-optimal operation on immunological status of EOC patients regarding also their overall survival (OS). On the day of primary cytoreduction and 7days after, the selected serum immunological parameters were determined in 49 patients with confirmed EOC. We found that, the level of immunosuppressive (interleukin 10; transforming growth factor-ß - TGF-ß1) and pro-inflammatory (interleukin-6 and 8) cytokines was significantly higher in the group of patients with advanced stage of disease, compared to early stage. However, the number of circulating CD3+, CD4+ or CD8+ cells, CD19+ and NK cells was similar in both group of EOC patients. The overall survival of patients who underwent optimal cytoreduction was significantly higher than that in whom only sub-optimal surgery was performed. Sub-optimal cytoreduction only partially weakened the serum level of TGF-ß1 and IL-8 and what is more enhanced the number of circulating CD4+CD25+high cells in patients with advanced stage of disease. Sub-optimal surgery and high post-operative level of TGF-ß1 increased the hazard ratio for patients. Besides, we noticed that the high pre-operative concentration of TGF-ß1 could distinguish all EOC patients (independently of FIGO classification) for whom optimal or sub-optimal surgery would be applied. Sub-optimal debulking resulted in higher immunosuppression and lower OS of EOC patients.

The methylenetetrahydrofolate reductase c.c.677 C>T and c.c.1298 A>C polymorphisms in reproductive failures: Experience from an RSA and RIF study on a Polish population.

Almost 1600 individuals from the Polish population were recruited to this study. Among them 319 were fertile couples, 289 were recurrent spontaneous abortion (RSA) couples, and 131 were in the group of recurrent implantation failure (RIF) following in vitro fertilization. The aim of this study was to evaluate the MTHFR c.c.677 C>T and c.c.1298 A>C polymorphisms' association with RSA and RIF. We used PCR-RFLP with HinfI (677 C>T) and MboII (1298 A>C) digestion. We observed a protective effect of the female AC genotype (OR = 0.64, p = 0.01) and the C allele (AC+CC genotypes; OR = 0.65, p = 0.009) against RSA. Moreover, 1298 AA/677 CT women were more frequent in RSA (31.14%) and RIF (25.20%) groups in comparison to fertile women (22.88%), although this difference was significant only in the case of RSA (p = 0.022, OR = 1.52). Male combined genotype analysis revealed no association with reproductive failure of their partners. Nevertheless, the female/male combination AA/AC of the 1298 polymorphism was more frequent in RSA couples (p = 0.049, OR = 1.49). However, the significant results became insignificant after Bonferroni correction. In addition, analysis of haplotypes showed significantly higher frequency of the C/C haplotype (1298 C/677 C) in the female control group than in the female RSA group (p = 0.03, OR = 0.77). Moreover, the association between elevated homocysteine (Hcy) level in plasma of RSA and RIF women and MTHFR polymorphisms was investigated but did not reveal significant differences. In conclusion, for clinical practice, it is better to check the homocysteine level in plasma and, if the Hcy level is increased, to recommend patients to take folic acid supplements rather than undergo screening of MTHFR for 1298 A>C and 677 C>T polymorphisms.

Possible Role of HLA-G, LILRB1 and KIR2DL4 Gene Polymorphisms in Spontaneous Miscarriage.

The KIR2DL4 receptor and its ligand HLA-G are considered important for fetal-maternal immune tolerance and successful pregnancy. The absence of a particular variant of KIR2DL4 might be a bad prognostic factor for pregnancy outcome. However, it could be compensated by the presence of the respective LILRB1 allele. Therefore, we investigated the KIR2DL4, LILRB1 and HLA-G polymorphisms in 277 couples with spontaneous abortion and 219 control couples by HRM, PCR-SSP and RFLP methods. We found a protective effect of women's heterozygosity in -716 HLA-G (p = 0.0206) and LILRB1 (p = 0.0131) against spontaneous abortion. Surprisingly, we observed more 9A/10A genotypes of KIR2DL4 gene carriers in the group of male partners from the miscarriage group in comparison to the men from the control group (p = 0.0288). Furthermore, there was no association of women's KIR2DL4 polymorphism with susceptibility to spontaneous abortion. Multivariate analysis indicated that women's -716 HLA-G and LILRB1 and men's KIR2DL4 9A/10A are important in terms of the protection or susceptibility to miscarriage, respectively (p = 0.00968). In conclusion, a woman's heterozygosity in HLA-G and LILRB1 might be an advantage for a success of reproduction, but the partner's heterozygosity in 9A/10A KIR2DL4 alleles might not.

Feasibility of implementation of a "simplified, No-X-Ray, no-lead apron, two-catheter approach" for ablation of supraventricular arrhythmias in children and adults.

INTRODUCTION: Although the "near-zero-X-Ray" or "No-X-Ray" catheter ablation (CA) approach has been reported for treatment of various arrhythmias, few prospective studies have strictly used "No-X-Ray," simplified 2-catheter approaches for CA in patients with supraventricular tachycardia (SVT). We assessed the feasibility of a minimally invasive, nonfluoroscopic (MINI) CA approach in such patients. METHODS: Data were obtained from a prospective multicenter CA registry of patients with regular SVTs. After femoral access, 2 catheters were used to create simple, 3D electroanatomic maps and to perform electrophysiologic studies. Medical staff did not use lead aprons after the first 10 MINI CA cases. RESULTS: A total of 188 patients (age, 45 ± 21 years; 17% <19 years; 55% women) referred for the No-X-Ray approach were included. They were compared to 714 consecutive patients referred for a simplified approach using X-rays (age, 52 ± 18 years; 7% <19 years; 55% women). There were 9 protocol exceptions that necessitated the use of X-rays. Ultimately, 179/188 patients underwent the procedure without fluoroscopy, with an acute success rate of 98%. The procedure times (63 ± 26 vs. 63 ± 29 minutes, P > 0.05), major complications (0% vs. 0%, P > 0.05) and acute (98% vs. 98%, P > 0.05) and long-term (93% vs. 94%, P > 0.05) success rates were similar in the "No-X-Ray" and control groups. CONCLUSIONS: Implementation of a strict "No-X-Ray, simplified 2-catheter" CA approach is safe and effective in majority of the patients with SVT. This modified approach for SVTs should be prospectively validated in a multicenter study.

Is there any association between secretory IgA and lactoferrin concentration in mature human milk and food allergy in breastfed children.

BACKGROUND: Breastfeeding is recommended as a protective method against the development of allergy. However, some studies have reported an increased risk of allergies development in breastfed infants of atopic mothers, which implies that atopic mothers may have an altered composition of breast milk. AIM: The aim of the study was to determine the concentration of secretory immunoglobulin A (S-IgA) and lactoferrin in human mature milk and to evaluate the association between the levels of these proteins in breast milk with food allergy in children, depending on the allergy status of the breastfeeding mother. MATERIAL AND METHODS: Medical data was collected from birth to 24 months of age from 84 mother-child pairs participating in an EU-funded project "EuroPrevall - The prevalence, cost and basis of food allergy across Europe". The diagnosis of food allergy in children was based on the positive result of a double-blind placebo-controlled food challenge (DBPCFC). S-IgA and lactoferrin levels were measured in the whey of mature breast milk with commercial enzyme-linked immunosorbent assay (ELISA) kits. Statistical analysis (the U Mann-Whitney and Kruskal-Wallis tests as well as the Spearman's rank correlation coefficient) was performed using STATISTICA 8.0 PL (Statsoft, Tulsa, USA). RESULTS: Ten out of eighty four participating children had positive skin prick tests (SPT) and/or sIgE to food antigens and in 7 (8.4%) DBPCFC confirmed food allergy. the median concentration of S-IgA was 476,83 µg/ml (range 6.51-1359.61 µg/ml). the median concentration of Lf was 15.68 µg/ml (range 11.68-36.43 µg/ml). The concentrations of S-IgA and Lf showed a moderate, negative, correlation R=-0.28; p=0.05. CONCLUSIONS: Mature breast milk of mothers of children with food allergy and of healthy children showed similar concentrations of both proteins. The level of S-IgA in the mature milk of mothers with atopic allergy was significantly lower, compared to non-atopic mothers. More studies are needed to reveal the mystery of the lack of protective effect of breastfeeding on allergy development in children.

[Can the concentration of immunomodulating factors in mature breastmilk be associated with food allergy in breastfed children?]. / Czy istnieje zwiazek miedzy stezeniem czynników immunomodulujacych w ludzkim mleku dojrzalym i wystapieniem alergii pokarmowej u dzieci karmionych piersia?

OBJECTIVES: The aim of the study was to determine the concentration of such immunomodulating factors as transforming growth factor beta1 (TGF-ß1), interleukin 10 (IL-10), tumor necrosis factor alpha (TNF-α) in mature human milk and to relate the levels of the above mentioned cytokines in mature breast milk to the occurence of food allergy in children during the first 24 months of life. MATERIALS AND METHODS: Data on breastfeeding, symptoms of food allergy in children and breast milk samples were collected prospectively from birth to 24 months of age from 84 mothers participating in the Polish birth cohort of "EuroPrevall" study, in the years 2005-2007. Cytokine levels were measured in the whey with commercial enzyme-linked immunosorbent assays (ELISA) kits. RESULTS: Ten out of the eighty four (11.9%) participating children had positive SPT and/or sIgE to food antigens. In 7 out of 84 (8.4%) children DBPCFC confirmed the diagnosis of food allergy. The median concentration of TGF-ß1 was 21.94 pg/ml (range 10.47-83.19), TNF-α 1.46 pg/ml (range 0.35-16.50), IL- 101.83 pg/ml (range 0.58-31.04). There was a positive correlation between the concentration of IL-10 and TGF-ß1. The level of TNF-α correlated positively with the duration of lactation (p=0.04). There was no significant difference between the concentration of IL-10, TGF-ß1, TNF-α, in the mature breast milk of mothers of children with symptoms of allergy and positive SPT and/or sIgE, mothers of children with positive DBPCFC and in the milk of mothers of control children. CONCLUSIONS: There was no significant difference between the concentration of IL-10, TGF-ß1, TNF-α, in the mature breast milk of mothers of children with food allergy and in the breast milk of mothers of control children.

Immunotherapy of patients with recurrent spontaneous miscarriage and idiopathic infertility: does the immunization-dependent Th2 cytokine overbalance really matter?

Recurrent spontaneous miscarriage (RSM) and idiopathic infertility (IIF) are partially caused by immunologic disturbances. Paternal lymphocyte immunization (PLI) is proposed for restoration of the proper Th1/Th2 balance in these patients, but still there are controversies on PLI mechanism, its efficacy and identification of patients who may benefit from this therapy. The study group consisted of n = 34 RSM and n = 42 IIF women with unexplained miscarriage or IIF. PLI was offered as a treatment in both groups. Peripheral blood lymphocyte (PBL) populations (CD3(+), CD3(-)/CD19(+), CD3(+)/CD4(+), CD3(+)/CD8(+), CD3(-)/CD16(+)CD56(+)) were studied before immunization, while PBL cytokine secretion (IFN-γ, TNF-α, IL-10, IL-5, IL-4, IL-2), before and after immunization, pre-conceptionally in both groups. The reference PBL ratio and cytokine levels were adopted from previously studied normal fertile women. PBL populations, concentration and ratio of Th1/Th2 cytokines did not differ between RSM and IIF patients. Compared to the results observed in normal fertile women the levels of IFN-γ, TNF-α and IL-2 were higher, while IL-10 lower in both RSM and IIF patients (p < 0.01). After immunization a decrease of IFN-γ (RSM and IIF groups) and IL-4 and IL-10 (RSM group) were observed, as well as an increase in TNF-α/IL-4 ratio (RSM group) (p < 0.05). No differences in Th1/Th2 concentration and ratio between patients with successful and unsuccessful pregnancy were observed. No significant correlations between success and particular cytokine concentration were observed. Concentrations of Th1/Th2 cytokines and PBL populations did not differ between RSM and IIF women. Th1 shift in both RSM and IIF patients was observed in comparison to fertile women. Treatment with PLI-induced pre-conceptionally cytokine changes which neither indicated Th2 shift nor correlated with subsequent pregnancy success.

HLA-C C1C2 heterozygosity may protect women bearing the killer immunoglobulin-like receptor AA genotype from spontaneous abortion.

Spontaneous abortion is the most common complication of human pregnancy. Natural killer (NK) cells expressing killer immunoglobulin-like receptors (KIRs), which may recognize HLA-C (i.e. its C1 or C2 groups) on trophoblast cells, constitute a large leukocyte population in the endometrium. This study investigated whether genetic polymorphisms in the KIR and HLA-C genes are risk factors for spontaneous abortion. One hundred and twenty-five couples with at least two spontaneous abortions, including eighty-five couples with idiopathic recurrent abortion (RSA; three or more abortions), and 117 control couples (with two or more healthy-born children) were tested. The frequencies of the individual KIR genes in the patients were similar to those in the controls. In the group of KIR AA women with HLA-C C2C2 partners, the HLA-C C1C2 heterozygotes were present in the controls but not in the patients (p=0.015 for all patients and p=0.0048 for RSA, but both comparisons lost significance after Bonferroni correction), whereas both homozygotes, C1C1 and C2C2, were absent in the control women but present among the aborting ones. Therefore, our results suggest that among KIR AA women who have HLA-C C2C2 partners, HLA-C heterozygous females show a trend towards an increased chance of successful pregnancy.

L-ficolin (ficolin-2) insufficiency is associated with combined allergic and infectious respiratory disease in children.

We previously reported an association between relative L-ficolin deficiency and recurrent respiratory infections co-existing with allergic disorders in children. To confirm and extend this preliminary finding, we performed a prospective study on children of a similar age (mean 8.9 years) designed to establish whether the principal relationship was with infection or allergy. Serum L-ficolin values in healthy children were normally distributed with a mean value of 3838 ng/ml. L-ficolin concentrations were generally lower in patients with asthma and/or allergic rhinitis with (mean 3413 ng/ml; p=0.02) or without (3512 ng/ml; p<0.07) respiratory infections, but not in patients with respiratory infections without allergic disease (3623 ng/ml; p=0.2). The lower average values in the group comprised of children with respiratory allergy and infections were largely due to a high proportion of very low values: 18.3% had values below 2150 ng/ml compared to only 5.5% of healthy controls (OR=3.9; p=0.01). This relationship was not apparent in the groups characterized by allergy without infection or infections without allergy. An association between mannan-binding lectin (MBL) insufficiency and recurrent respiratory infections was also confirmed. One of the patients was MASP-2 deficient, evidenced both by MASP2 genotyping and by lectin pathway activity measurement. In conclusion, L-ficolin may confer some protection from microorganisms that exacerbate allergic inflammation in the lung and its relative deficiency may contribute to enhanced susceptibility to respiratory infections. MBL insufficiency and MASP-2 deficiency are risk factors for recurrence of infections independently of allergic disease.

Effect of nitric oxide donors on NADPH oxidase signaling pathway in human neutrophils in vitro.

The production of reactive oxygen species (ROS) in activated neutrophils is catalyzed by NADPH oxidase, a multiproteins complex. Various protein kinases including protein kinase C (PKC) and mitogen activated protein kinases (MAPKs) are involved in NADPH oxidase phosphorylation and activation. The main step in the NADPH oxidase activation is phosphorylation of its cytosolic protein p47(phox). We found previously that nitric oxide (NO) donors such as metabolite of molsidomine-SIN-1 and diethylamine/NO significantly impaired the ROS production in activated human neutrophils. In this study, we investigated the effects of both NO donors on NADPH oxidase-linked signaling proteins in activated neutrophils. We found that NO donors decreased the phosphorylation of p47(phox) on tyrosine and serine/threonine residues and PKC on serine residues in neutrophils. Both NO donors did not affect the phosphorylation of MAPKs. NO donors partially but significantly lost their ability to reduce ROS production in the presence of PKC but not MAPKs inhibitors. We show that whereas NO donors have no effect on MAPKs activity, they do decrease PMA- and/or fMLP-induced phosphorylation of p47(phox) and PKC as well as PMA- and fMLP-induced ROS production.

Isolated head injury in children affects the neutrophil function and lymphocyte count.

BACKGROUND: Outcomes of treatment of postinjury complications remain unsatisfactory and research continues into the impact of trauma on innate and acquired immunity. The aim of our study was to describe how head injury affects a child's immunity by measuring the neutrophil function and lymphocytes subsets. METHODS: The peripheral blood of 16 children with head trauma (Glasgow Coma Score < or =9) was examined. The blood samples were collected on the first and on the seventh day after trauma. The production of reactive oxygen species (ROS), spontaneous and stimulated, the expression of CD11b, and the lymphocyte subpopulations were measured. The blood of healthy children was studied as control. The impact of endotracheal intubation on the examined parameters was analyzed as well. RESULTS: Head trauma leads to the increase of leukocytosis; the total production of ROS by peripheral blood neutrophils does not change after head injury. Correction of the results according to the number of neutrophils revealed a significant decrease in ROS production by a single neutrophil. The expression of adhesion molecule CD11b did not change. Head injury in children causes the decrease of the total lymphocyte count, CD3, CD4, CD8, and natural killer cells count on both the first and the seventh postinjury day. On the seventh day the significant decrease of natural killer cells subset was observed. The CD4/CD8 ratio increased from 1.5 (the first day) to 2.5 (the seventh day). The intubation did not affect the examined parameters. CONCLUSIONS: After head injury, total ROS production and adhesion molecule CD11b expression remained unchanged when compared with control. The study did not demonstrate evidence for neutrophil activation in patients with head injuries. The total lymphocyte count was found to be decreased and the composition of lymphocytes' subsets was deeply impaired.

[Assessment of postvaccinal response in children past pertussis]. / Ocena odpowiedzi poszczepiennej u dzieci, które przebyly krztusiec.

In the nineties, despite high percentage of vaccination of children, from 86% to 99% dependently on the region, there came to the increase of whooping cough cases. Until then infants and children to the age of 5 years suffered from this disease, whereas in the last decade the number of cases increased mainly among children over 5 years of age, who were subjected to full cycle of vaccination against whooping cough. Searching for the causes of such epidemiologic situation the following suggestions have been given: change of the bacteria antigenicity, the phenomenon of postvaccinal immunity extinction, immune system mechanisms disorders and groundless excuse from vaccination. The study estimates parameters of postvaccinal response to immunization with selected vaccines in children post whooping cough.

Influence of systemic photochemotherapy on regulatory T cells and selected cytokine production in psoriatic patients: a pilot study.

BACKGROUND: Psoriasis is a chronic autoimmune inflammatory disease of the skin with strong genetic and environmental risk factors and is regarded as a Th1 cell-type disease. The aim of our study was to evaluate the effect of one-month PUVA (Psoralen Ultraviolet A) therapy on a regulatory T-cell subpopulation (CD4+CD25+) and the production of some cytokines. MATERIAL/METHODS: The study was performed on the group of 12 patients with severe psoriasis. They were put on PUVA therapy for one month. We analyzed the level of CD4+CD25+ regulatory T cells using a FACSCalibur cytometer and CellQuest Software. The production of IFN-gamma (interferon-gamma), TNF-alpha (tumor necrosis factor alpha), IL (interleukin) -10, IL-5, IL-4, and IL-2 by lymphocytes was estimated by using a CBA system. The control group consisted of 11 healthy volunteers. RESULTS: We found that the production of INF-gamma, TNF-alpha, IL-2, and IL-10 in psoriatic patients before PUVA application increased significantly compared with the control group. In patients after PUVA therapy we observed decreased production of TNF-alpha and a decreased number of CD4+CD25+ cells in the blood compared with the same group of patients before the treatment. CONCLUSIONS: It was demonstrated that systemic PUVA therapy led to a marked reduction in CD4+CD25+ T cells and a change in cytokine production.

[Alterations of procalcitonin and interleukin 6 after cardiopulmonary bypass in children with congenital heart disease]. / Zmiany stezen prokalcytoniny i interleukiny 6 u dzieci z wrodzonq wadq serca podczas zabiegu kardiochirurgicznego i po jego zakonczeniu.

UNLABELLED: Cardiac surgery induces systemic inflammatory response that may have been implicated the postoperative organ dysfunction. This inflammatory response is thought to be produced by exposing patients to proinflammatory factors. The aim of our study was to investigate alterations in procalcitonin (PCT) concentration in peripheral blood in children as the potential early indicator of complications occurring during and after surgery in extracorporeal circulation. Additionally, we evaluated the perioperative time course of IL-6. MATERIAL AND METHODS: The investigations were carried out in 21 children undergoing cardiac operation with cardiopulmonary bypass (CPB). Serum concentrations of PCT and IL-6 were sequentially measured before induction of anesthesia, at the initiation of CPB, at the end of CPB, and 24 hours, and 72 hours after CPB. RESULTS: There was no significant PCT-elevation at all 5 times of measurement. Levels of IL-6 increased significantly after surgery, and remained elevated for up to 1 day. Peak values correlated with the duration of CPB (r=0.68, p=0.0006). CONCLUSIONS: We conclude, that cardiac surgery with CPB did not have any influence on the secretion of PCT in children. These results suggest that IL-6 was more effective than PCT to monitor patients with a favorable outcome.

[Effect of high doses of shark liver oil supplementation on T cell polarization and peripheral blood polymorphonuclear cell function]. / Modyfikujacy wplyw duzych dawek preparatu oleju z watroby rekina na polaryzacje limfocytów T i funkcje neutrofili krwi.

Fish oils supplementation has been recently widely used in prevention and treatment of the diseases in humans. Fish oil beneficial effects have been investigated in a number of animal disease models as well as human studies. Here, we examined clinical, immunological and biochemical effects of shark liver oil supplementation in high doses in 13 volunteers. The experiment was based on the consumption of 3.6 g of squalene, 3.6 g of alkylglycerols and 750 mg of n-3 polyunsaturated fatty acids (PUFA) per day for 4 weeks. We have shown the increased response of neutrophils towards bacteria, the increased level of C4 component of complement in blood, the rise of total antioxidant status of serum, and the predominance of Type I cytokine IFN-gamma, TNF-alpha and IL-2 production by peripheral blood mononuclear cells after shark liver oil intake. Moreover, shark liver oil supplementation markedly affect lipid metabolism and cholesterol balance. The increase of total cholesterol level from 182.92 +/- 29.290 mg/dl before oil consumption to 224.46 +/- 62.198 mg/dl after diet rich in oil, and the decrease of HDL fraction were noted. However, metabolism of lipids normalised spontaneously after the end of the experiment in all the individuals. The results of the present study have shown, that the main effects of shark liver oil are the result of the biological activity of squalene and 1-O-alkylglycerols, which dominate in the composition of the oil quantitatively. On the contrary, anti-inflammatory effects of n-3 PUFA do not manifest, when taking together with high doses of squalene and alkylglycerols. On the bases of these observations, we propose that shark liver oil supplementation in high doses is beneficial in bacterial, viral and fungal infections, whereas patients with atherosclerosis or autoimmune diseases should avoid the consumption of high amounts of shark liver oil.

Flow cytometric evaluation of human neutrophil apoptosis during nitric oxide generation in vitro: the role of exogenous antioxidants.

Among numerous inflammatory mediators a nitric oxide molecule is supposed to be important in the modulation of neutrophil survival in vivo and in vitro. The effect of exogenous supply of NO donors such as SNP, SIN-1, and GEA-3162 on the course of human neutrophil apoptosis and the role of extracellular antioxidants in this process was investigated. Isolated from peripheral blood, neutrophils were cultured in the presence or absence of NO donor compounds and antioxidants for 8, 12, and 20 hours. Apoptosis of neutrophils was determined in vitro by flow cytometric analysis of cellular DNA content and Annexin V protein binding to the cell surface. Exposure of human neutrophils to GEA-3162 and SIN-1 significantly accelerates and enhances their apoptosis in vitro in a time-dependent fashion. In the presence of SNP, intensification of apoptosis has not been revealed until 12 hours after the culture. The inhibition of GEA-3162- and SIN-1-mediated neutrophil apoptosis by superoxide dismutase (SOD) but not by catalase (CAT) was observed. Our results show that SOD and CAT can protect neutrophils against NO-donors-induced apoptosis and suggest that the interaction of NO and oxygen metabolites signals may determine the destructive or protective role of NO donor compounds during apoptotic neutrophil death.

Predominance of Type 1 cytokines and decreased number of CD4(+)CD25(+high) T regulatory cells in peripheral blood of patients with recurrent aphthous ulcerations.

Recurrent aphthous ulcerations (RAU) are a chronic inflammatory disease with evidence of inappropriate immune response. Previous studies have suggested cell-mediated activation of immune response towards common micro-organisms of oral cavity in RAU. In this investigation, we explored cytokine production by peripheral blood mononuclear cells (PBMC) and T regulatory cell population in blood of active and remission RAU patients as crucial factors for maintenance of peripheral tolerance. Ten patients with minor RAU and 12 healthy individuals were selected for the study. Cytokine levels were analysed in supernatants using Cytometric Bead Array Kit for flow cytometry and ELISA. We have demonstrated increased production of Type 1 cytokines IL-2, IFN-gamma and TNF-alpha as well as IL-5, IL-6 and IL-8 by peripheral blood mononuclear cells in RAU. In contrast, IL-10 and TGF-beta anti-inflammatory cytokine production was decreased in RAU patients compared to healthy individuals. Moreover, we have found that CD4(+)CD25(+high) T regulatory cell proportion was decreased in RAU and represented 3.58+/-0.654% of CD4(+) T cells in active RAU, 4.66+/-0.561% of CD4(+) T cells in remission RAU, whereas in healthy controls CD4(+)CD25(+high) T cells represented 7.30+/-1.238% of CD4(+) T cells (p<0.001). Thus, the obtained results indicate that disproportion in cytokine production may be contributing factor in the pathogenesis of RAU. Alteration in the number of CD4(+)CD25(+high) T regulatory cells in RAU may additionally influence the development of the disease. We propose that imbalance in pro- and anti-inflammatory cytokine network may lead to the breakdown of peripheral tolerance in RAU and the excessive immune response towards harmless micro-organisms colonized oral mucosa or self-antigens.

Cytokines locally produced by lymphocytes removed from the hypertrophic nasopharyngeal and palatine tonsils.

OBJECTIVE: Human palatine tonsils and the nasopharyngheal tonsil are the largest components of the Waldeyer's ring. Subepithelial and intraepithelial lymphocytes of human adenoids and tonsils are responsible for the local and the systemic immune response. We studied the cytokine production by lymphoid cells isolated from 16 nasopharyngeal tonsils (adenoid) and 9 palatine tonsils surgically removed by from 25 children (aged from 4 to 15 years) suffering from tonsil hypertrophy. METHODS: We evaluated (by the cytometry method, using BD Bioscience kits, San Diego, CA) the concentration of IL-2, IL-4, IL-5, IL-10, TNF(alpha) and IFN(gamma) released from human peripheral blood mononuclear cells (MC) (activated or not activated by phytohaemagglutinin (PHA)) cultured in vitro during 72 h. The fluorescence-activated cell sorter (FACS) analysis was also performed and the percentage of mononuclear cells (unstimulated or activated by phorbol acetate during 24 h) stained with the monoclonal antibodies anti-CD3 containing the intracellular cytokines was calculated. RESULTS: The increased secretion of IL-2, IL-4, IL-5, TNF(alpha) and IFN(gamma) from PHA activated palatine origin immune cell cultures, as compared to adenoids, was revealed. The higher mobilization (Delta%) of CD3+ T-lymphocytes containing IL-12 in palatine cell cultures (798.5+/-276.29%), in comparison with to the adenoids (298.5+/-49.16%; p< or =0.05), was also noted. CONCLUSION: In palatine tonsils, as compared to adenoids, the cellular immune (Th1) response dominates over humoral immune (Th2) reaction.

The involvement of immunoregulatory T cells in the pathogenesis of lichen sclerosus.

BACKGROUND: The pathogenesis of lichen sclerosus (LS) and the mechanism of involution of LS-affected tissues is controversial. Autoimmune factors are proposed as a cause of disease. The involvement of autoimmune T lymphocytes in the disease development and progression is considered. MATERIAL/METHODS: The investigation included 41 woman divided into two groups: a study group and controls. In the study group were 22 vulvular LS patients. Nineteen healthy woman underwent plastic surgery of the same area and served as controls. We analyzed reactive oxygen intermediate (ROI) production by peripheral blood granulocytes, the production of IL-2, IL-5, IL-10, IL-12, and TNF-alpha cytokines by lymphocytes, and the activation of CD25, CD26, CD69, CD71, and HLA DR antigen expression. RESULTS: An increase in CD4+CD25+ suppressor T cells together with a decrease in CD3+CD26+ activated lymphocytes paralleled an increase in IL-10 production by peripheral blood lymphocytes of the lichen sclerosus patients. Diminished ROI production by peripheral blood granulocytes of LS patients was observed after both receptor-dependent and -independent stimulation. Baseline increase in IL-12 and stimulated increases in IL-2, IL-5, IL-10, and TNF-alpha production by lymphocytes of LS patients were also observed. CONCLUSIONS: The involution of lichen sclerosus-affected tissues may be the suppressive effect exerted by CD4+CD25+ suppressor T lymphocytes, the increase in IL-10 inhibitory cytokine production, and diminished granulocyte ROI production. Inflammatory infiltrates in the affected regions of the skin are characterized by a diminished number of CD3 lymphocytes bearing the CD26 molecule, which may be responsible for an autocrine defect in bioactive mediator degradation.

Epidermal growth factor enhances TNF-alpha-induced priming of human neutrophils.

The intensity of neutrophil inflammatory response could be rapidly amplified by priming with pro-inflammatory mediators such as TNF-alpha, GM-CSF or LPS at low concentrations prior to stimuli. We proposed that epidermal growth factor (EGF) increases TNF-alpha-induced priming of human neutrophils. This study showed that EGF enhanced TNF-alpha-induced activation of neutrophils functions. The addition of EGF to neutrophils cultured with TNF-alpha resulted in increased respiratory burst and phagocytic activity of polymorphonuclear leukocytes (PMN) and up-regulation of adhesion molecule CD11b. Moreover, EGF enhanced IL-8 production by TNF-alpha-primed PMN. EGF alone was able to prime CD11b expression and IL-8 production by PMN. EGF receptor selective tyrosine kinase inhibitor, tyrphostin AG-1517, blocked the effect of priming with EGF, whereas the status of non-primed and TNF-alpha-primed neutrophils remained unaffected. EGFR expression on neutrophils was confirmed by flow cytometry and CELISA methods. These data provide the original evidence that EGF significantly enhances TNF-alpha-induced priming of human neutrophils acting through EGFR tyrosine kinase pathway. The observed effect may be a result of co-operative action of EGF, TNF-alpha and reactive oxygen intermediates (ROI).