Expression of toll-like receptors on human rectal adenocarcinoma cells.

The innate immune system uses Toll-like receptors (TLR) to detect the presence of pathogen patterns thus allowing for rapid host defense responses. Stimulation of TLR results in inflammatory response and regulatory cytokine production affecting acquired immunity. The aim of the study was an evaluation of TLR2 and TLR4 expression on the surface of human colon cancer cells in primary culture with or without autologous peripheral blood mononuclear cells. Surgical specimens of colon cancer were processed to obtain cancer cells. Cancer cells separation was conducted first by mechanical tissue disintegration and than by gradient centrifugation to obtain 95 % cell confluence. By staining the isolated cells the pathologist determined them as adenocarcinoma. Colon cancer cells were then co-cultured in 24 h culture alone or together with autologous lymphocytes. Reverse-transcription polymerase chain reaction was performed for detection of TLR2 and TLR4 mRNA in colon cancer and normal colon epithelial cells using commercially available primers. Resting as well as phytohemagglutinin or lipopolysaccharide (LPS) stimulated cells were tested. Receptor proteins on cancer cells were examined by immunohistochemistry. TLR4 mRNA was detected in cancer cells. Autologous lymphocytes do not exert any effect on these receptors expression. TLR4 mRNA expression was not observed in normal colon epithelial cells. TLR2 mRNA was present on LPS stimulated cancer cells as well as on resting and stimulated lymphocytes. Expression of TLR2 and TLR4 receptor proteins on colon cancer cells were confirmed by immunohistochemistry. TLR4 may be responsible for uncontrolled tumor growth under LPS stimulation in human colon environment.

[Neutrophils and hyaluronians in carcinogenesis]. / Neutrofile i hialuroniany w chorobach nowotworowych.

Inflammatory processes involving polymorphonuclear leukocytes (PMNL) play an important role in cancer. PMNL can affect the formation and growth of the tumor. Cancer cells produce significant amounts of hyaluronans (HA), which pass into the extracellular space and are responsible for the inhibition of PMNL function through TLR4 receptor, and increase in the production of IL 10 the immunosuppressive cytokine as well as the other proinflammatory cytokines. HA supported the metastasis of cancer cells by binding to the CD44 receptor on endothelial cells. Increased PMNL--Lymphocyte ratio as a negative prognostic factor indicates also the important not defined role of PMNL in carcinogenesis.

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.

[Anti-tumor activity of 1-O-alkylglycerols--the main component of shark liver oil]. / Wybrane aspekty dzialania przeciwnowotworowego 1-O-alkilogliceroli--glównej komponenty oleju z watroby rekina.

Fish oils contain several active compounds that modify cell activity and influence various functions of the human body. Shark liver oils are rich in 1-O-alkylglycerols which have strong ability to stimulate human immune system. In this review we discuss findings of the recent studies that showed antitumor properties of 1-O-alkylglycerols derived from fish oils and its effect in adjunctive treatment of several types of cancer.

Expression of Th1/Th2/Th3/Th17-related genes in recurrent aphthous ulcers.

The pathogenesis of recurrent aphthous ulceration (RAU) is unknown, although an abnormal immune reaction appears to be involved. RAU may result from oral epithelium damage caused by T cell-mediated immune response. To improve understanding of the role of T cells in RAU, the present study analyzed. the expression of T cell-related genes in oral ulcers from patients with RAU, as well as in healthy non-keratinized oral mucosa from aphthae-free volunteers. Biopsies from RAU patients and healthy individuals were analyzed using Human Th1-Th2-Th3 RT(2) Profiler PCR Array and qRT-PCR that allowed to quantify the transcript levels of 86 genes related to T cell activation. We found that cells present in aphthous ulcers express a characteristic Th1-like gene profile. The majority of genes up-regulated in aphthous lesions such as IFN-γ, TNF, IL-15, IRF1, STAT-1 and STAT-4 were Th1-associated. Th2-realated genes were not overexpressed in RAU tissues, with the exception for CCR3. Th3- and Th17-related gene expression patterns were not demonstrated in RAU. These findings clearly reveal that aphthous ulcer formation is predominantly dependent on the activation of the Th1-type immune response.

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.

Evaluation of TLR4 expression and chosen parameters of oxidative-antioxidative balance in young children with food allergy.

The authors evaluated mRNA TLR4 expression on neutrophils and the chosen parameters of oxidative-antioxidative balance in blood of 35 children with food allergy (17 of them with IgE-dependent allergy and 18 with IgE-independent allergy) and 15 healthy children without any allergy. The age of these children ranged from 1 to 36 months. Children with food allergy in comparison with healthy children were found to have lower mRNA TLR4 expression, higher average value of chemiluminescence (CL) and its increase after stimulation by fMLP, PMA and OZ as well as lower TAS values. Disturbances of oxidative-antioxidative balance were found in children with food allergy. We suggest that natural immunity is involved in the development of food allergy mechanisms. Moreover, chemiluminescence can be used as an additional diagnostic test.

Does the KIR2DS5 gene protect from some human diseases?

BACKGROUND: KIR2DS5 gene encodes an activating natural killer cell receptor whose ligand is not known. It was recently reported to affect the outcome of hematopoietic stem cell transplantation. METHODOLOGY/PRINCIPAL FINDINGS: In our studies on KIR2DS5 gene associations with human diseases, we compared the frequencies of this gene in patients and relevant controls. Typing for KIR2DS5 gene was performed by either individual or multiplex polymerase chain reactions which, when compared in the same samples, gave concordant results. We noted an apparently protective effect of KIR2DS5 gene presence in several clinical conditions, but not in others. Namely, this effect was observed in ankylosing spondylitis (p=0.003, odds ratio [OR]=0.47, confidence interval [CI]=0.28-0.79), endometriosis (p=0.03, OR=0.25, CI = 0.07-0.82) and acute rejection of kidney graft (p=0.0056, OR=0.44, CI=0.24-0.80), but not in non-small-cell lung carcinoma, rheumatoid arthritis, spontaneous abortion, or leukemia (all p>0.05). In addition, the simultaneous presence of KIR2DS5 gene and HLA-C C1 allotype exhibited an even stronger protective effect on ankylosing spondylitis (p=0.0003, OR=0.35, CI=0.19-0.65), whereas a lack of KIR2DS5 and the presence of the HLA-C C2 allotype was associated with ankylosing spondylitis (p=0.0017, OR=1.92, CI=1.28-2.89), whereas a lack of KIR2DS5 and presence of C1 allotype was associated with rheumatoid arthritis (p=0.005, OR=1.47, CI=1.13-1.92). The presence of both KIR2DS5 and C1 seemed to protect from acute kidney graft rejection (p=0.017, OR=0.47, CI=0.25-0.89), whereas lack of KIR2DS5 and presence of C2 seemed to favor rejection (p=0.0015, OR=2.13, CI=1.34-3.37). CONCLUSIONS/SIGNIFICANCE: Our results suggest that KIR2DS5 may protect from endometriosis, ankylosing spondylitis, and acute rejection of kidney graft.

Proinflammatory and immunosuppressive serum, ascites and cyst fluid cytokines in patients with early and advanced ovarian cancer and benign ovarian tumors.

OBJECTIVE: To analyze the profiles of interleukin-2 (IL-2), IL-6, IL-8, IL-10, tumor necrosis factor-alpha (TNF-alpha), transforming growth factor-beta1 (TGF-beta1) and interferon-gamma (IFN-gamma) in serum and the tumor microenvironment (cyst fluid, ascites) in women with ovarian cancer or benign ovarian tumors to find the differences in their immunological status. We also estimated serum cytokines as biomarkers to distinguish preoperatively between malignant or benign character of tumors. DESIGN: Prospective study. SETTING: Tertiary referral hospital. POPULATION: 51 women with epithelial ovarian cancer, 26 with benign ovarian tumors of epithelial origin and 21 healthy controls. METHODS: The levels of cytokines were measured using ELISA sets. RESULTS: We did not found differences in the levels of IFN-gamma, TNF-alpha and IL-2 in all fluids isolated from patients with malignant or benign tumors. Women with advanced cancer had significantly higher serum IL-6, IL-10 and TGF-beta1 levels than women with early stages or benign tumors. Moreover, women with very advanced cancer in whom the optimal cytoreduction was disabled had the highest serum levels of IL-10, TGF-beta1 and IL-8. The concentrations of IL-6 and IL-8 were higher in ascites of cancer patients than in ascites of women with benign tumors. The areas under curves constructed for the selected cutoff serum cytokines levels (AUC-ROC) showed good predictive values for IL-6 (0.87), IL-10 (0.836) and IL-8 (0.797). CONCLUSIONS: Our results indicate on intensified inflammatory process in women with ovarian cancer (accompanied by their immunosuppression). Preoperative analysis of serum IL-6, IL-10 and IL-8 may improve the differential diagnosis of ovarian tumors.

Antioxidant effect of hyaluronan on polymorphonuclear leukocyte-derived reactive oxygen species is dependent on its molecular weight and concentration and mainly involves the extracellular space.

INTRODUCTION: Hyaluronan (HA), a component of the extracellular matrix, may regulate immune cell functions through its interactions with cellular receptors. Besides its effect on cytokine and chemokine production, its antioxidant properties have been described. However, the mechanisms of this are not fully elucidated. The aim of this study was to evaluate the relationship between HA concentration and molecular weight and its antioxidant properties towards human neutrophils. Also assessed was whether the antioxidant effect of HA is connected with a reduction in intracellular oxygen potential, which could indicate its direct effect on neutrophil respiratory burst. MATERIALS/METHODS: The relationship between HA's antioxidant properties and its concentration and molecular weight was assessed by the luminol-enhanced chemiluminescence method (CL). To evaluate the effect of HA on intracellular oxygen potential selectively, the dihydrorhodamine 123 (DHR123) flow cytometric method was used. RESULTS: Reduction of both HA molecular weight and its concentration decreased its antioxidant properties in the CL method. A selective effect of HA on intracellular oxygen potential measured by the DHR123 method was not shown. CONCLUSIONS: The antioxidant properties of HA are related to both its molecular weight and its concentration. The lack of an antioxidant effect of HA in the DHR123 test compared with a significant reduction in CL values at the same HA concentration suggests that HA acts mainly as a chemical ROI scavenger in the extracellular space.

[Production of proinflammatory cytokines by peripheral blood mononuclear cells in cocultures with squamous cell carcinoma of the larynx]. / Wydzielanie cytokin prozapalnych przez jednojadrzaste komórki krwi obwodowej w hodowli z komórkami raka plaskonablonkowego krtani.

PURPOSE: Antitumor mechanisms of cellular immune response are connected with the cytokines activity secreted by peripheral blood mononuclear cells (PBMC). The aim of this study was estimation of proinflammatory cytokine (IL-6 and TNF- ) concentration in patients with laryngeal squamous cell carcinoma and analysis of directed influence of neoplasm cells to the function of PBMC and modification of cytokine profile. MATERIALS AND METHODS: For analysis of cytokine secretion, the cultures of isolated peripheral blood mononuclear cells (PBMC) with marginal neoplasm cells and noncancerous adjacent mucosa cells from 55 patients with laryngeal squamous cell carcinoma were established. Production of cytokines in 21h and 72h culture supernatants was detected by Elisa. RESULTS: Authors reported the increase of IL-6 and decrease of TNF- concentration in cultures of isolated peripheral blood mononuclear cells (PBMC) with marginal neoplasm cells and non-cancerous adjacent mucosa cells. CONCLUSION: The presence of laryngeal squamous carcinoma cells in PBMC culture can modify immunocompetent cells activity and production of proinflammatory cytokines.

Dysfunction of CD4+CD25high T regulatory cells in patients with recurrent aphthous stomatitis.

BACKGROUND: Recurrent aphthous stomatitis (RAS) is a chronic inflammatory disease of unknown etiology characterized by recurring formation of painful oral ulcers. RAS may result from oral epithelium damage caused by T-cell-mediated immune response. CD4(+)CD25(+) T regulatory (Treg) cells suppress proliferation and effector functions of other immune cells, and therefore are crucial in regulating the immune response. METHODS: We tested the function of peripheral CD4(+)CD25(high) Treg cells in active RAS through their ability to inhibit proliferation and cytokine production of conventional CD4(+) T cells. We also attempted to detect the presence of FOXP3 and indoleamine 2,3-dioxygenase (IDO) mRNA in the lesional and non-lesional oral mucosa of RAS patients and healthy individuals using real-time PCR assay. RESULTS: Treg cells derived from RAS patients were less efficient in the suppression of cytokine production of CD4(+) T effector cells than Treg cells from healthy individuals. Moreover, in RAS, Treg cells were nearly twice less potent in the inhibition of CD4(+)CD25(-) T cell proliferation than in healthy donors. Furthermore, we have demonstrated the decreased proportion of CD4(+)CD25(+)FOXP3(+) Treg cells in peripheral blood of RAS patients compared with controls. We failed to detect FOXP3 mRNA, while IDO mRNA expression was decreased in non-lesional mucosa biopsies from RAS patients compared with ulcer biopsies or normal mucosa from healthy donors. CONCLUSIONS: These findings suggest that CD4(+)CD25(high) Treg cells are both functionally and quantitatively compromised in RAS and that decreased constitutive expression of IDO in oral mucosa in RAS may lead to the loss of local immune tolerance.

[Hyaluronan-mediated regulation of inflammation]. / Hialuronian jako czynnik regulujacy proces zapalenia.

Hyaluronan is high-molecular-weight biopolymer. Its linear structure is created by repeating disaccharide units. A single unit is composed of N-acetyl-D-glucosamine and D-glucuronic acid. Hyaluronan is the main component of the extracellular matrix. Apart from its structural role, hyaluronan can influence cell proliferation, differentiation, and migration, angiogenesis, as well as inflammation and immune cell function. During inflammation, high-molecular-weight hyaluronan is broken down under the influence of free radicals and enzymes to smaller fragments. Numerous literature data show that the effect of haluronan on immune cells is closely dependent on its molecular mass. High-molecular-weight hyaluronan can participate in restraining inflammation, while the low-molecular-weight form possesses a proinflammatory effect and activates immune cells. Through interaction with surface receptors (CD44, RHAMM, TLR4), hyaluronan fragments stimulate immune cells and enhance cytokine and reactive oxygen species production as well as other factors participating in inflammation. Hyaluronate can thus be an important regulator of the inflammatory process. Low-molecular-weight fragments deliver signal about tissue damage and mobilize immune cells, while the high-molecular-form suppresses immune cell function and prevents excessive exacerbation of inflammation.

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.

Lipopolysaccharide-activated CD4+CD25+ T regulatory cells inhibit neutrophil function and promote their apoptosis and death.

CD4+CD25+ T regulatory (Treg) cells play a central role in the suppression of immune response and prevention of autoimmune reactions. Pathogen recognition receptors expressed by immune cells, such as TLRs, may provide a critical link between the innate and adaptive immune systems. There is also evidence that TLR ligands can directly modulate the suppressive capacity of Treg cells. Here, we showed that CD4+CD25+ Treg cells affect neutrophil function and survival and that the TLR4 ligand is involved in the regulation of the cell interactions. We found that LPS-activated Treg cells inhibit reactive oxygen intermediates and cytokine production by neutrophils. Moreover, Treg cells reverse LPS-induced survival of neutrophils and promote their apoptosis and death. We also found that TCR-activated Treg cells induce the same effects on polymorphonuclear neutrophils as those achieved by TLR4 stimulation. Importantly, the suppressive potential of CD4+CD25+ Treg cells induced by LPS seems to be partially IL-10 and TGF-beta dependent, whereas anti-CD3/CD28 stimulation is rather contact dependent. Together, these observations suggest that Treg cells have the ability to directly regulate neutrophil function and life span when both types of the cells are exposed to LPS.

[Inflammation is a key effector phase in immune reactions]. / Zapalenie podstawowa reakcja efektorowa w zjawiskach immunologicznych.

Inflammation is key element in effector phase of all immune processes and create a cascade system of specific enzyme functions regulated in autocrine way. Humoral antigen-antibody reaction activate the complement system which is responsible for secondary mediatots generation and tissues destruction. Liberated mediators like kinins, leukotriens inflammatory cytokines and activated cell bound metalloproteases in tissues and blood vessels are responsible for oedema of tissues, pain, cell destruction and organ dysfunction. Induction of T cell dependent inflammatory reaction involve antigen dependent activation of T cells which usually is delayed and long lasting. The immune reactions responsible for specific activation induce inflammation.

[Biological action and clinical application of shark liver oil]. / Mechanizm dzialania i zastosowanie kliniczne oleju z watroby rekina.

Fish oils contain several active compounds that modify cell activity and influence various functions of the body. Shark liver oils are rich in alkylglycerols and squalene, but contain relatively low amounts of n-3 polyunsaturated fatty acids. Alkylglycerols may control immune response possibly throw modification of platelet activating factor (PAF) and diacylglycerol (DAG) production. Squalene enhances antigen presentation and induction of inflammatory response. Moreover, alkylglycerols and squalene have antitumour activity, that is possibly based on different mechanisms, ie., induction of apoptosis of neoplastic cells, suppression of signal transduction, inhibition of angiogenesis and promoting of transmembrane transport of cytotoxic agents. Shark liver oil has been found to be useful in treatment of conditions resulted from inadequate immune response, and in adjunctive treatment of several types of cancer.

Interleukin 6 and 8 levels in plasma and fibroblast cultures in psoriasis.

Fibroblasts have been implicated in psoriatic inflammatory processes. The aim of the study was to evaluate soluble interleukin 2 receptor (sIL-2R), interleukin 6 (IL-6), and interleukin 8 (IL-8) plasma levels in psoriatic patients and IL-6 and IL-8 levels in fibroblast culture supernatants. Cytokines levels in plasma and supernatants were measured by ELISA. Plasma sIL-2R, IL-6, and IL-8 levels were higher before the treatment in comparison to healthy controls (P < 0.001) and decreased after treatment. Fibroblasts from healthy controls, psoriatic lesional skin, and noninvolved psoriatic skin, when stimulated with tumor necrosis factor alpha, released considerable amounts of IL-6 and IL-8. No significant difference between healthy controls and psoriatic fibroblasts was observed. Monitoring plasma sIL-2R levels could be employed as a reliable method of psoriasis activity. IL-8 and IL-6 plasma levels seem to reflect psoriasis activity, and treatment response, respectively. Fibroblasts are not a major source of increased IL-6 and IL-8 production in psoriasis.

[Does the hearing loss affect the child's innate immunity?--preliminary]. / Niedosluch a odpornosc naturalna dziecka? (doniesienie wstepne).

UNLABELLED: Human neutrophiles play a crucial role in inmate immunity. Inmate and aquired immune response depend on their functional condition at the first stage of an inflammation process. Reactive forms of oxygen (RFO) produced by neutrophiles play a significant role in the eradication of pathogens as well as in the regulation of immune response. The aim of this study is question, does the hearing loss affect the inmate immunity? METHOD: The RFO production was directly examined in four systems - without stimulation, after stimulation with fMLP, opsonized zymosan or PMA. Direct RFO measurement was performed chemiluminescency evaluation using the whole blood, which indirectly depend on RFO production. RESULT: In the group of children with hearing loss was observed disturbances in RFO productions. CONCLUSION: This observation is very original and important for general practice.

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.