Guanosine may increase absence epileptic activity by means of A2A adenosine receptors in Wistar Albino Glaxo Rijswijk rats.

The non-adenosine nucleoside guanosine (Guo) was demonstrated to decrease quinolinic acid(QA)-induced seizures, spontaneously emerged absence epileptic seizures and lipopolysaccharide(LPS)-evoked induction of absence epileptic seizures suggesting its antiepileptic potential. It was also described previously that intraperitoneal (i.p.) injection of 20 and 50mg/kg Guo decreased the number of spike-wave discharges (SWDs) in a well investigated model of human absence epilepsy, the Wistar Albino Glaxo Rijswijk (WAG/Rij) rats during 4th (20mg/kg Guo) and 3rd as well as 4th (50mg/kg Guo) measuring hours. Guanosine can potentially decrease SWD number by means of its putative receptors but absence epileptic activity changing effects of Guo by means of increased extracellular adenosine (Ado) cannot be excluded. An increase in the dose of i.p. injected Guo is limited by its low solubility in saline, therefore, we addressed in the present study whether higher doses of Guo, diluted in sodium hydroxide (NaOH) solution, have more potent antiepileptic effect in WAG/Rij rats. We confirmed that i.p. 50mg/kg Guo decreased but, surprisingly, i.p. 100mg/kg Guo enhanced the number of SWDs in WAG/Rij rats. Combined i.p. injection of a non-selective Ado receptor antagonist theophylline (5mg/kg) or a selective Ado A2A receptor (A2AR) antagonist SCH 58261 (7-(2-phenylethyl)-5-amino-2-(2-furyl)-pyrazolo-[4,3-e]-1,2,4-triazolo[1,5-c]pyrimidine) (1mg/kg) and a cyclooxygenase 1 and 2/COX-1 and COX-2 inhibitor indomethacin (10mg/kg) with 100mg/kg Guo decreased the SWD number compared to i.p. 100mg/kg Guo alone. The results suggest that i.p. 100mg/kg Guo can increase SWD number by means of the adenosinergic system.

Antiphospholipid antibodies in relation to sterility/infertility.

Of the systemic autoimmune diseases that lead to sterility/infertility, antiphospholipid syndrome (APS) has an outstanding importance; it may be associated with abortion and premature birth which are included in its diagnostic criteria, as well as preeclampsia, pregnancy-induced hypertension, foetal retardation, miscarriage, stillbirth and sterility. Between 2004 and 2009, on the Department of Immunology in the Zala County Hospital, 100 female patients with sterility (st)/infertility (if) (33/67), (mean age: 34.08 years) underwent, in addition to history taking and physical examination, an assessment by immune-serologic tests (ANA, anti-dsDNA, ENA-Profile, anti-TPO, a-sperm, aCL, aPS, aß2GP1, aANX, and aPT). Positive aCL on two occasions could be demonstrated in 27/100 cases (27%) (st/if: 7/20). Among them 4 cases of primary APS have been diagnosed respectively. In the remaining 17 patients the clinical picture did not fulfil criteria. In addition to the twofold positive aCL, unusual antiphospholipid antibodies including aß2GP1, aPS or both were present in 1/27, 2/27 and 1/27 patients, as well as aANX and aPT in 3/26 and 1/27 patients respectively. One-time positive aCL occurred in 16/100 women (16%) (st/if: 5/11); among them aPT and aß2GP1 could be detected in 1/16 patient each. Based on the clinical picture, we raised the possibility of primary APS in 2/16 patients. Among the aCL-negative women, we found the unusual antibodies of APS in 8/57 patients (14%) including positivity of aß2GP1, aPS, aPT and aANX in 4/57, 4/57, 2/57 and 3/57 patients respectively; taking the clinical criteria of APS into consideration, primary APS could be stated in 2/57 patients of them. The 32 pregnancies developed in the follow-up period upon administration of acetylsalicylic acid (ASA) and maintenance dose low molecular weight heparin (LMWH), together with prednisolone in patients with secondary APS, resulted in 23 deliveries and 5 miscarriages; 4 pregnancies are currently in progress. The results of our investigations highlight the significance of demonstrating latent autoimmune diseases in female patients with sterility/infertility, as barrenness can be terminated by the appropriate treatment of these conditions.

Combined plasmapheresis and high-dose intravenous immunoglobulin treatment in systemic sclerosis for 12 months: follow-up of immunopathological and clinical effects.

Systemic sclerosis (SSc) is an autoimmune disease which involves the skin, as well as several internal organs. Most therapies available in this disease are symptomatic. Authors present a case of diffuse SSc with progressive disease not responding to currently available treatments. Therefore a 12-month protocol of repeated plasmapheresis and high-dose intravenous immunoglobulin treatment was administered with good clinical efficacy. Apart from monitoring the clinical symptoms throughout the treatment, authors also assessed a number of humoral and cellular immunolaboratory markers in order to obtain information on the immunomodulatory effects of this combined treatment in SSc.

Intracytoplasmic cytokine expression and T cell subset distribution in the peripheral blood of patients with ankylosing spondylitis.

OBJECTIVE: To determine the role of inflammatory mediators in the pathogenesis of ankylosing spondylitis (AS), we investigated peripheral blood lymphocyte subsets and their intracellular cytokine production. METHODS: The percentages of T and B lymphocytes, natural killer (NK) cells, activated T lymphocytes, CD4+ T helper (Th), and CD8+ T cytotoxic (Tc) cells were determined by flow cytometry in 42 patients with AS compared to 52 healthy controls. In order to assess circulating Th1/Th2 and Tc1/Tc2 subsets, we used a whole-blood cytometric assay based on the intracellular interferon-gamma, interleukin 4 (IL-4), and IL-10 expression of the cells. RESULTS: In the peripheral blood, the frequencies of CD4+ T helper and CD56+ NK cells were higher in AS (54.8% and 16.2%, respectively) compared to controls (45.3% and 10.8%) (p < 0.05). The frequencies of Th0 (1.9% vs 0.8%) and Tc0 (2.1% vs 0.8%) cells were higher, while that of Tc1 cells was lower (26.6% vs 40.1%) in patients with AS versus controls (p < 0.05). The percentage of IL-10-producing Tc cells was significantly higher in AS (18.4%) versus controls (8.5%) (p < 0.05). Finally, the active phase of AS was associated with significantly lower percentage of IL-10-producing Tc cells in the peripheral blood (6.6%) compared to patients with inactive AS (23.1%). CONCLUSION: Our results provide further evidence for an altered T cell subset distribution and intracytoplasmic cytokine balance in AS.

Progressive divergent shifts in natural and induced T-regulatory cells signify the transition from undifferentiated to definitive connective tissue disease.

The objectives of the study is to determine clinical signs and distribution of peripheral T-cell subsets, B cells and T regulatory cells in patients with undifferentiated connective tissue disease (UCTD) and during the development toward well-established connective tissue diseases (CTD). The methods include 46 patients with UCTD were followed and investigated for differentiation into defined CTDs for 2 years. Cell subsets were determined on the basis of cell surface markers, intracellular cytokine production by flow cytometry and serum cytokine levels by ELISA. The results are as follows: 45.6% of UCTD patients developed into a defined CTD. The number and percentage of activated T cells, memory T cells and NKT cells were increased in patients compared with controls. In addition, in patients with UCTD, the percentage of CD4+/IFN gamma+ T(h)1 was significantly higher compared with controls and further increased in patients that developed CTDs. The percentage and absolute number of CD4+CD25+Foxp3+ regulatory T cells (Tregs) were diminished in UCTD patients compared with healthy controls, while the number of CD4+/IL-10+ Tregs increased. The conclusions are Overproduction of IFN gamma and the decrease of natural (Foxp3+) Tregs seem to be characteristic features of UCTD patients. The increased IL-10 production of CD4+ T cells might be a compensatory suppressive mechanism; however, it is probably not able to balance the overproduction of IFN gamma and the observed decrease of Foxp3+ Tregs. The shift toward T(h)1 with increased IFN gamma production in patients with UCTD combined with the degree of immunoregulatory disturbances characterized by the progressive divergent shifts in natural and induced T-regulatory cell populations signify the transition from undifferentiated to definitive CTD.

Associations between serum anti-CCP antibody, rheumatoid factor levels and HLA-DR4 expression in Hungarian patients with rheumatoid arthritis.

BACKGROUND: The presence of anti-cyclic citrullinated peptide autoantibody is highly specific for rheumatoid arthritis. Certain HLA-DR4 (HLA-DRB1*04) alleles, also known as the "shared epitope," are associated with increased susceptibility to RA. In addition, these alleles may also have relevance for disease outcome. Anti-CCP antibody positivity has been associated with the presence of HLA-DR4 alleles in patients with RA. However, there is little information regarding a relationship between quantitative anti-CCP production (serum anti-CCP concentrations) and the shared epitope. OBJECTIVES: To determine the association between anti-CCP antibody production and various HLA-DRB1 alleles. METHODS: Serum anti-CCP, rheumatoid factor and C-reactive protein levels were assessed in 53 RA patients. All these patients underwent HLA-DRB1 genotyping. RESULTS: Of the 53 patients 33 (62%) were positive for anti-CCP antibody. We found significant correlations between anti-CCP and RF positivity (chi-square = 6.717, P < 0.01), as well as between anti-CCP and HLA-DRB1*04 positivity (chi-square = 5.828, P < 0.01). There was no correlation between RF positivity and serum levels, CRP serum levels and HLA-DRB1*04 positivity. When quantitatively comparing serum anti-CCP levels with shared epitope positivity, patients carrying one or two copies of HLA-DRB1*04 alleles had significantly higher anti-CCP concentrations (530.0 +/- 182.6 U/ml) compared to DRB1*04-negative patients (56.8 +/- 27.4 U/ml) (P < 0.01). There was no difference in serum anti-CCP antibody concentrations between patients carrying only one HLA-DRB1*01 allele but no HLA-DRB1*04 allele (12.0 +/- 8.6 U/ml) compared to SE-negative patients (76.8 +/- 56.2 U/ml). Regarding non-SE HLA-DRB1 genotypes, all 6 patients (100%) carrying DRB1*15 alleles and 6 of 7 (85%) patients carrying DRB1*13 were anti-CCP positive. In addition, patients with HLA-DRB1*13 (282.5 +/- 23.8 U/ml) and DRB1*15 (398.7 +/- 76.2 U/ml) produced significantly more anti-CCP than did any other non-SE HLA-DRB1 subtypes (P < 0.01). CONCLUSIONS: There is significant association between anti-CCP and RF, as well as between anti-CCP and SE positivity in RA. In addition, the presence of one or two copies of HLA-DRB1*04 alleles has been associated with higher serum anti-CCP antibody levels. Thus, patients carrying HLA-DRB1*04 alleles exhibited an overall tenfold increase in serum anti-CCP antibody levels in comparison to HLA-DRB1*04-negative subjects. Increased anti-CCP production may also be associated with other non-SE HLA-DRB1 genotypes, such as DRB1*13 or DRB1*15. In reports by other investigators, both anti-CCP concentrations and SE positivity were related to more rapid disease progression and unfavorable outcome.

Immunoregulatory T cells in the peripheral blood of patients with Hodgkin's lymphoma.

We aimed at investigating the distribution of various types of immunoregulatory T cells in the peripheral blood of patients with Hodgkin's lymphoma (HL) (n = 94) being in the state of long-lasting complete remission using flow cytometry. Healthy patients (n = 41) as 'negative' and patients in complete remission with breast cancer (n = 47) as 'positive' controls were investigated in the study. We found significant elevations in the number of CD4+ CD25high naturally occurring regulatory T cells, CD4+/intracellular IL-10+ (Tr1) and CD8+/intracellular IL-10+ T cells in HL compared to the healthy controls. In carcinoma patients, however, the number of Tr1 and CD8+/IL-10+ T cells was higher than that in the other two groups. The increase in the number of CD4+ CD25high T cells seems to be characteristic of HL compared to the two other types of regulatory T cells. This change exists for a long time and it seems to be a characteristic of HL and independent of the types of therapy and the duration of time since therapy.

Cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted mice.

BACKGROUND: Our department worked out a modified surgical form of spleen autotransplantation earlier, named "spleen apron method" introduced already into the clinical practice. Recently we tested the immunological changes in a group of patients autotransplanted with about 10-15% of their spleen, what was the at least always implantable amount after the severe splenic injuries. In the current work we aimed at measuring some cellular and serological changes in the peripheral blood of splenectomized and spleen autotransplanted inbred mice two and eight months after the operations in order to get more unambiguous results than that we could obtain in our patients with this technique. MATERIALS AND METHODS: We divided 96 two months old Balb/c female mice into eight groups (n = 12/group). The group of controls, sham operated, splenectomized and autotransplanted animals with two and eight months of survival time after the operations. During the autotransplantation we inserted the same amount of spleen, five slices, "chips," about 10-15% of total mass of spleen, into the greater omentum similarly as it was used in the patients. The concentration of serum proteins were measured by laser nephelometry. The lymphocyte subsets were analyzed by flow cytometry. RESULTS: We found that two months after the operations the number of CD 19+ B-cells increased in the splenectomized but decreased in the autotransplanted animals. Eight months after the operations the number of both CD3+ T and CD19+ B lymphocytes decreased both in the splenectomized and autotransplanted animals compared to the controls and sham operated mice. However, the numbers of T and B cells were slightly but not significantly higher in the autotransplanted than in the splenectomized mice. The serum level of IgM was also decreased in the splenectomized and autotransplanted mice at both time points, however, eight months after the operations the concentration of IgM was significantly higher in the autotransplanted group than in the splenectomized animals. CONCLUSION: The effects of autotransplanted "chips" were different at the various ages of the animals. Additionally, they showed some immunological benefit being quantitatively in accordance to the amount of the transplanted spleen. The elevated level of serum IgM what we found in the autotransplanted mice even with this amount of transplanted spleen eight months after the operations, however, might have the potentially greatest importance compared to splenectomy. These experiments can prove that the attempts for autotransplantation may have real perspectives but their efficacy depends on the amount of the successfully transplanted (saved) mass of spleen.

Glucocorticosteroid therapy decreases CD14-expression and CD14-mediated LPS-binding and activation of monocytes in patients suffering from systemic lupus erythematosus.

In order to study the possible action of glucocorticosteroids (GCS) on the CD14/Toll like receptor mediated activation of monocytes the CD14-expression, CD14-mediated LPS binding and activation of these cells of patients suffering from Systemic Lupus Erythematosus receiving no, low dose or pulse steroid treatment was studied. The CD14-expression was determined on whole blood monocytes by flow cytometry, while the LPS-binding of an FITC-LPS preparate and the LPS-induced TNFalpha secretion were tested on isolated monocytes. The CD14-dependent and -independent LPS-binding and activation were evaluated with the help of a blocking anti-CD14 mAb. Our results showed that the CD14-expression, CD14-dependent LPS-binding and activation were significantly inhibited by the in vivo applied pulse steroid therapy. In contrast, the CD14-independent LPS-binding and activation were not altered by the GCS treatment. Our data provide further in vivo evidence for a possible new way of GCS therapy is able to initiate its anti-inflammatory action.

[Comparative hematologic and immunologic studies of patients with splenectomy and spleen autotransplantation]. / Splenectomizált és lép autotranszplantált betegek összehasonlító haematologiai és immunologiai vizsgálatai.

We compared changes in haematological and immunological parameters of patients with splenectomy (n=24), splenectomy plus spleen autotransplantation (n=12) and healthy controls (n=23). In the autotransplantation group pieces of the removed spleen were placed into the omentum with good visible circulation. Significant alterations in the hematological status and in some immunological parameters were observed in both groups of patients who were operated on compared to those in the control group. There was no difference, however, between the results of the two groups of operated patients. Therefore, we emphasize the importance of vaccination in patients with spleen autotransplantation in order to prevent potential sepsis. In addition, we recommend the possible further use of spleen autotransplantation.

Can CD3+/HLA-DR+ activated T cells predict the prognosis of non-Hodgkin's lymphoma patients?

The immune system has several mechanisms to fight against developing malignant cell clones in the host, one of them is the activated T-cell response. Both CD4+ helper and CD8+ cytotoxic T-cells bear HLA-DR molecules as important surface activation markers. Our aim was to determine, how the ratio of activated T cells change in the peripheral blood of non-Hodgkin's lymphoma (NHL) patients during the periods of polychemotherapy. Using the methods of immunofluorescence staining and flow cytometry, we found the ratio of CD3+/HLA-DR+ cells significantly higher in NHL patients before treatment compared to healthy controls (10.63% versus 2.97%, P<0.001). During the period of polychemotherapy, this ratio began to increase significantly (10.63% versus 16.94%, P=0.006). After treatment, the ratio of activated T cells decreased, however, we detected significantly higher rate of CD3+/HLA-DR+ lymphocytes in patients who relapsed within 1 year than in those who stayed in remission (9.55% versus 20.62%, P<0.001). We suppose that investigation of CD3+/HLA-DR+ activated T cells might be a promising method to determine the prognostics of lymphoma patients.

Intracellular IL-4/IFN-gamma producing peripheral T lymphocyte subsets in B cell non-Hodgkin's lymphoma patients.

AIM: The availability of several biological therapy options is rapidly growing in the field of malignant lymphomas. This emphasizes the need to understand the precise interaction of the host immune system with the malignant disease. We measured the intracellular cytokine responses in lymphoma patients' lymphocytes, to characterize the polarization changes in their immune system. METHODS: Patients with B cell non-Hodgkin's lymphoma were involved in the study. Peripheral lymphocytes were labeled with anti-CD4 or anti-CD8 antibodies and intracellular accumulation of IL-4 or IFN-gamma was detected after in vitro incubation the cells with activating cocktail. The frequency of different T-cell subsets were measured within the lymphocyte population. RESULTS: Significantly increased CD4+, IFN-gamma producing (Th1) cell percentage were found in untreated lymphoma cases (28.8% vs. 21.8%). CD8+ IL-4 and IFN-gamma producing (Tc0) T cell frequency is significantly higher in untreated lymphoma patients compared with normal controls (1.3% vs. 0.47%). The frequency of CD4+ IL-4 producing (Th2) cells is significantly lower in untreated patients (0.96% vs. 1.19%). Patients in long-term remission have lower frequency of CD4+, IL4 producing (Th2) cell ratio (0.31% vs. 1.19%) and increased CD4+ IFN-gamma producing (Th1) cell frequency (30.1% vs. 21.8%), compared with healthy normal controls. CONCLUSION: Our results demonstrate that there is a sustained increase in the CD4+ IFN-gamma producing cell frequency in lymphoma patients. The frequency of CD4+ IL-4 producing cells is decreasing by treatment. These may contribute to strong polarization toward Th1 type response, needed for lymphoma clearance and remission.

Relationship between skin bacterial colonization and the occurrence of allergen-specific and non-allergen-specific antibodies in sera of children with atopic eczema/dermatitis syndrome.

In this study we investigated skin bacterial colonization, allergen-specific IgE and antiphospholipid/antinuclear antibodies in 72 children with atopic eczema/dermatitis syndrome (age 2-17 years). Bacteria were found on the skin in 41 cases and serological allergen-specific IgE positivity in 37. The different forms of antibodies appeared in the ratio 21/72 (33 antibodies in 21 children). The occurrence of antiphospholipid antibodies was significantly higher in the patients than in the controls. There were significantly more allergens in the group with bacterial colonization than in the group without colonization. The SCORAD index showed a significant positive association with the skin colonization. We conclude that there are significant relationships between the occurrence of Staphylococcus aureus colonization and the levels of inhalant allergen-specific IgE in children with atopic eczema/dermatitis syndrome, and between the occurrence of antiphospholipid IgM positivity and atopic eczema/dermatitis syndrome.

[Plasmapheresis therapy in systemic sclerosis]. / Plazmaferezis-kezelés szisztémás sclerosisban.

BACKGROUND: Systemic sclerosis (SSc) is a polysystemic autoimmune disorder characterized by excessive production of extracellular matrix and vascular abnormalities, involving the skin and internal organs. There is currently no real disease-modifying therapy of SSc and there are only few reports on the use of plasmapheresis in SSc. OBJECTIVES: Authors present 4 cases of SSc treated with plasmapheresis within one year from disease onset. All patients had diffuse cutaneous form of SSc with rapid progression and with different internal organ manifestations. METHODS: The patients were treated with plasmapheresis performed every 3 months besides symptomatic drug treatment. Clinical symptoms, Rodnan skin score, immunolaboratory markers (serum immunoglobulin, complement, several autoantibodies, leukocyte CD markers) and soluble adhesion molecules (E-selectin, intercellular adhesion molecule 1, vascular cell adhesion molecule, which were showed as the markers of disease activity in previous works) were determined before and after the plasmapheresis. RESULTS: The progression of the disease slowed down significantly in all patients, no more new clinical manifestation appeared, Rodnan skin score significantly decreased. There was a reduction in CD4/CD8 ratio (approaching normal ratio), in expression of CD69, as early activation marker. There was a significant suppression of soluble adhesion molecules also in all patients. CONCLUSION: In patients with diffuse cutaneous form of SSc, plasmapheresis may be effective with decreasing progression of the disease and improving clinical symptoms (especially skin manifestation) in the early phase of the disease.

Elevated rate of Thelper1 (T(H)1) lymphocytes and serum IFN-gamma levels in psoriatic patients.

Several disorders are known to be associated with altered Thelper1/Thelper2 (T(H)1/T(H)2) cytokine balance. Psoriasis is characterized by increased systemic and local production of T(H)1 and pro-inflammatory cytokines. Furthermore recent data indicate the dominant presence of T(H)1 lymphocytes in the circulation and T(H)1 and Tcytotoxic1 (T(C)1) cells in lesional skin of psoriatic patients. In order to assess the systemic T(H)1/T(H)2 imbalance in psoriasis most of the studies so far tested isolated peripheral mononuclear cells. As a new approach we applied a whole blood flow cytometric assay to determine the rate of circulating T(H)1/T(H)2 and T(C)1/Tcytotoxic2 (T(C)2) lymphocytes based on their intracellular IFN-gamma, IL-4 and IL-10 expression. Besides, serum levels of these cytokines were determined in healthy controls and psoriatic patients by commercial ELISAs. In psoriatic patients we found significantly (P<0.02) increased rates of CD4(+)/IFN-gamma(+) lymphocytes (30.3+/-8.8%) while the percent of CD4(+)/IL-4(+) cells (0.37+/-0.31%) were significantly (P<0.03) lower compared to healthy controls (CD4(+)/IFN-gamma(+): 20.1+/-7.3% and CD4(+)/IL-4(+): 0.78+/-0.44%). The IL-10-positive CD4(+) and CD8(+) cells also had higher rate in psoriasis, but the difference between patients and controls was not significant, similarly to the rate of CD8(+)/IFN-gamma(+) and CD8(+)/IL-4(+) lymphocytes. Beside cellular expression, serum IFN-gamma levels were also significantly higher (control: 4.9+/-6.4 pg/ml; psoriatic patients: 35.9+/-47.0 pg/ml; P<0.05). Our results provide further evidence for an altered T(H)1/T(H)2 balance in psoriasis measured in non-separated whole blood T cells.

No effect of transfusion transmitted virus viremia on the distribution and activation of peripheral lymphocytes in hemodialyzed patients.

AIM: We aimed to examine the distribution and activation of peripheral T cells in TTV positive (n = 32) and negative (n = 17) hemodialyzed patients. The control group (n = 20) consisted of healthy blood donors. METHOD: TTV-DNA was detected by seminested PCR. CD3, CD4, CD8, CD19, CD56, CD3/HLA-DR, CD3/CD69 and the Th1/Th2 ratio of T cells were analyzed by flow cytometry. Circulating IFN-gamma, IL-2, IL-4, IL-6, IL-10, IL-13, TNF-alpha, TGF-beta levels were measured by ELISA in the sera. RESULTS: There was no difference between the CD3, CD4, CD8 and CD19 values of HD subjects. In addition, the expression of both activation markers, HLA-DR and CD69, was significantly elevated in the TTV-positive and -negative HD groups compared to the controls, but not showing any difference from each other. The measurements of intracellular cytokines showed the enhanced occurrence of INF-gamma + CD4 T cells, and decreased appearance of IL-4 + CD4 lymphocytes in the HD groups without any significant difference between the TTV virus positive and negative patients. In addition, HD also elevated the expression of IL-10 in CD4 and CD8 (Th2) cells. There were only two significant changes in the levels of circulating cytokines: (a) IL-2 increased; (b) IL-13 decreased in both groups of HD patients compared to the controls, independently of TTV positivity or negativity. CONCLUSIONS: We assume that transfusion-transmitted virus does not cause any specific change in the distribution and activation of lymphocytes in the peripheral blood of hemodialyzed patients. Hemodialysis itself, however, results in a significant activation of peripheral T cells with the domination of increased production of Th1 type cytokines, IFN-gamma, IL-2, in contrast to the decreased synthesis of Th2 type cytokines, IL-4 and IL-13. Furthermore, the increased expression of IL-10 in the CD4 and CD8 cells of HD patients can be the sign of a contraregulatory Th2 activation as an answer on the Th1 effect.

Serum cytokine levels and type 1 and type 2 intracellular T cell cytokine profiles in mixed connective tissue disease.

OBJECTIVE: To determine serum cytokine concentrations and intracellular cytokine production of CD4+ and CD8+ T cells in 20 patients with mixed connective tissue disease (MCTD). METHODS: Detailed analysis of cytokine production; 8 patients were in the active stage, 12 in the inactive stage of the disease. Serum concentrations of interferon-gamma (IFN-gamma), interleukin 4 (IL-4), and IL-10 were assessed by ELISA. Intracellular content of IFN-gamma, IL-4, and IL-10 in CD4+ and CD8+ peripheral blood T cell and lymphocyte subsets was determined by flow cytometry. RESULTS: Serum concentrations of both type 1 and type 2 cytokines were significantly higher in patients with MCTD than in healthy controls. IFN-gamma expression of CD4+ and CD8+ T cells did not differ comparing all patients to controls. In patients with active MCTD, the percentage of CD8+/IFN-gamma+ Tc1 cells was significantly increased compared to inactive disease or to controls (p < 0.05). IL-4 expression of CD4+ T cells was scarcely detectable in MCTD, while a subpopulation of CD8+ Tc2 cells produced IL-4. A higher percentage of these CD8+/IL-4+ Tc2 cells were detected in patients with MCTD, especially in active disease, compared to controls (p < 0.05). The percentage of IL-10-expressing CD4+ and CD8+ T cells was higher in patients than in controls (p < 0.05). Again, CD4+ and CD8+ T cells from patients with active MCTD produced significantly more IL-10 than cells in patients with inactive disease or in controls (p < 0.05). CONCLUSION: Our results suggest that MCTD is associated with increased production of both type 1 (IFN-gamma) and type 2 cytokines (IL-4, IL-10). Cytokine production is usually higher in active MCTD than in inactive disease. CD8+ T cells may produce more IFN-gamma and IL-10 in comparison to CD4+ T cells. Patients with active disease have more IL-4-expressing Tc2 cells and IL-10-expressing Th2 and Tc2 cells than patients with inactive MCTD or controls. In MCTD, increased IL-10 production by Th2 and Tc2 cells may be an attempt by the immune system to downregulate the inflammatory reaction, although this effect may not be sufficient to restore the physiological Th1/Th2 balance in MCTD.

Cytokine production of CD4+ and CD8+ peripheral T lymphocytes in patients with chronic idiopathic urticaria.

The aim of this study was to investigate the characteristic cytokine pattern of patients with chronic idiopathic urticaria. Using flow cytometry, we examined the frequency of IL4, IL-10, IL-13 and IFN-gamma producing CD4+ and CD8+ T cells in the peripheral blood mononuclear cells at a single cell level. In patients with chronic idiopathic urticaria, the frequency of IL-10 producing CD4+ and CD8 + T cells was significantly higher than that of control subjects, while the frequency of IFN-y producing helper and cytotoxic T cells was significantly lower. The proportion of IL-4 producing CD4 + T cells from patients with urticaria was significantly lower. The ratio of IL-4 producing CD8 + T cells and the proportion of IL-13 producing CD4 + and CD8 + T lymphocytes did not show any significant difference between patients and controls. In our study, we could observe neither a dominant Th1 nor a dominant Th2 type cytokine pattern. We found a significant elevation in the intracellular IL-10 level which may be the cause of the down-regulated Th1 and Tc1 and partly Th2 lymphocyte functions.

[Does the TT virus affect T-cell subgroups in patients undergoing hemodialysis?]. / Hatással van-e a TT-vírus a hemodializált betegek T-sejt alcsoportjaira?

BACKGROUND: Recently many publications have appeared about the new DNA virus, called transfusion transmitted virus (TT virus), first described in 1997. These are mainly about the virus epidemiology, gene sequences and the distribution of different genotypes. In spite of the fact that the prevalence of this type of infection can reach 40 percent rate in polytransfused patients, such as in hemodialysis patients, the real pathogenetic effect of the virus has not yet been known. AIMS: The aim of the authors was to examine the activation and distribution of mononuclear cells in peripheral blood and to analyse the possible changes in Th1/Th2 immune regulatory mechanism through the soluble and intracellular cytokine profile beside the biochemical parameters of hepatic lesions in TT virus positive (n = 32) and negative (n = 17) hemodialysed patients. Healthy blood donors were the control group (n = 20). METHOD: Semi-nested PCR was used to detect the DNA of TT virus. For the surface antigen (CD3, CD4, CD8, CD19, CD56, CD3/HLA-DR, CD3/CD69) and intracellular cytokine analysis the authors applied flow cytometric method. RESULTS: The authors did not find any differences in the liver specific biochemical parameters between TT virus positive and negative hemodialysed and the healthy control group. The number of total T, T helper and total B cells were decreased. The percentage of CD8+, CD3+/HLA-DR+, CD3+/CD69+ and CD56+ cells were increased significantly in both hemodialysed population independently the presence or absence of TT virus. The soluble and intracellular cytokines showed significant growth of the Th1/Th2 cells ratio in hemodialysed patients, which has not been modified by the virus. CONCLUSIONS: From these results the authors assume that the TT virus does not cause any significant changes in the immune regulation, although it could play some role in the pathogenesis of hepatitis by local reaction.

[Amyopathic dermatomyositis]. / Amyopathiás dermatomyositis.

Dermatomyositis is a systemic autoimmune disease which belongs to the group of idiopathic inflammatory myopathies. The disease is rare with an incidence of 0.1-1/100,000 and a prevalence of 1-6/100,000. Women are affected twice as often as men. In some patients the disease presents with dermatologic changes weeks to months before the myopathy arises. It was observed that in some patients the myositis develops much later and sometimes not at all. Therefore, the term of dermatomyositis sine myositis used in earlier literature has been changed to amyopathic dermatomyositis. The most important question is whether the patient needs systemic therapy with its possible side effects yet possibly preventing the appearance of myositis or only local therapy for the skin manifestations is necessary. The goal of this article is to summarize the latest findings in amyopathic dermatomyositis.