Expression of nucleic acid binding Toll-like receptors in control, lupus and transplanted kidneys--a preliminary pilot study.

We hypothesized that nucleic acids, free and/or complexed, filtered and/or locally released, might be entrapped in the kidneys because of the specific nucleic acid binding microbial pattern recognizing Toll-like receptors (TLRs). This hypothesis of nucleic acid binding potential was tested using paraffin sections from healthy control, SLE and transplant kidneys, which were labelled using TLR-specific rabbit or goat anti-human antibodies in immunoperoxidase staining. Normal and transplant kidneys contain some double- (TLR-3) and single-stranded RNA binding (TLR-8) receptors, but in particular double-stranded RNA binding receptor TLR-7, mostly in tubuli, whereas no DNA binding TLR-9 was found. SLE kidneys contain more TLR-3 and TLR-8 and express de novo also TLR-9, in particular in glomeruli. On the contrary, TLR-7 was relatively weak in SLE. It is concluded that kidneys have a capacity to bind nucleic acids. TLR stimulation leads to the production of tumour necrosis factor-alpha and other pro-inflammatory cytokines and to up-regulation of co-stimulatory molecules necessary for the adaptive immune response. This makes renal tissues a potential target for inflammatory and immune responses in autoimmune disease and in the reaction for the foreign tissue.

Immunological profiles of patients with chronic myeloid leukaemia. I. State before the start of treatment.

In view of the increasing interest in the immunotherapy of CML it seems highly desirable to broaden the present knowledge on the immune reactivity of CML patients. A group of 24 patients and 24 healthy controls were studied for the total of 15 immunological parameters, including the prevalence of antibodies against human herpesviruses and papillomaviruses. To clearly discriminate between changes associated with the disease and those induced by the therapy, all patients were enrolled prior to the start of any anti-leukaemic therapy. Statistically significant differences between patients and controls were found in the levels of IgA, C4 component of complement, CRP and IL-6, the production of Th1 cytokines in stimulated CD3 cells and the E. coli stimulatory index. The analysis of the interrelationship between the results obtained in the individual patients presented some unexpected findings, such as the lack of correlation between the CRP and IL-6 levels. It will be the purpose of a follow-up to determine whether and how the immune status of the patients prior to the treatment correlates with their response to therapy and how the individual immunological profiles change in the course of the disease. These observations will be utilized in the future immunotherapeutic studies to constitute the vaccine- and placebo-treated groups.

C1q complement component and -antibodies reflect SLE activity and kidney involvement.

The role of the complement system in the pathogenesis of systemic diseases is very ambivalent. In systemic lupus erythematosus (SLE), many abnormalities in the activation of the complement system have been reported. The most important antibodies formed against the complement system in SLE are the ones associated with the C1q component. The aim of this study was to assess separately the anti-C1q antibodies and C1q component in the serum from 65 patients with SLE, then in individuals with (n=33) and without (n=32) lupus nephritis and with active (n=36) and nonactive (n=29) form of the disease (European Consensus Lupus Activity Measurement, ECLAM>3, ECLAM

Serum levels of the MCP-1 chemokine in patients with ischemic stroke and myocardial infarction.

Chemokine-driven migration of inflammatory cells has been implicated in pathogenesis of atherosclerosis-associated conditions such as ischemic stroke and myocardial infarction. In this study, a candidate chemokine, monocyte chemoattractant protein (MCP)-1, was investigated in patients with both aforementioned manifestations of atheroslerotic inflammation. MCP-1 levels in serum were determined by ELISA in 40 healthy, control subjects (C), 40 patients with ischemic stroke (IS), and in 64 patients with myocardial infarction (MI). Statistical analysis utilised Mann-Whitney test, Fisher's exact test, and Spearman's rank correlation (P < .05). In comparison to control subjects (C; median/interquartile range: 239/126 pg/mL), MCP-1 serum levels were increased in both investigated patient cohorts (IS: 384/370, P < .001; MI: 360/200, P < .002). There was a substantial variability of MCP-1 serum levels, especially in the IS group. No relationship was observed between chemokine levels and atherosclerosis risk factors (hypertension, diabetes, smoking, and alcohol consumption), and MCP-1 was also not related to age or gender. Elevation of MCP-1 in circulation of patients with atherosclerosis-associated complications implicates this CC chemokine ligand (CCL)2 in inflammatory processes, which contribute to pathogenesis of myocardial infarction and ischemic stroke. Further investigations, including patient stratification, are however necessary to evaluate if MCP-1 can be utilised for clinical management of patients with these diseases.

[Neopterin and a soluble interleukin-2 receptor in patients with systemic lupus erythematodes]. / Neopterin a solubilní receptor interleukinu-2 u nemocných se systémovým lupus erythematodes.

Goal of this study was to monitor levels of serum neopterin and soluble interleukin-2 receptor (sIL-2r) and to evaluate their importance in monitoring activity of systemic lupus erythematodes (SLE). Levels of serum neopterin, anti-dsDNA antibodies, C3, C4 complement components, nucleosomes antibodies, IL-10, fas ligand, soluble thrombomodulin, sVCAM-1, and sICAM-1 were measured in a group of 52 patients with SLE. Positive correlations were proved between neopterin concentrations and disease activity (ECLAM), levels of sVCAM-1, sICAM-1, sIL-2r and thrombomodulin, further between sIL-2r level and disease activity (ECLAM), and concentrations sVCAM-1, sICAM-1 and neopterin. Higher values of neopterin and sIL-2r levels were identified in patients with lupus nephritis compared to patients without kidney impairment. Statistically significant differences were identified in levels of neopterin between a subgroup (A) with minimum disease activity and a subgroup (B) with increasing disease activity (p = 0.01) and a subgroup (C) with decreasing disease activity (p = 0.003 ) and a subgroup (LN) with lupus nephritis (0.007) during the first and the third series of measurements. sIL2r levels which had in all subgroups very varied values were the lowest in the subgroup A with minimum disease activity during the whole time of monitoring. The highest levels reached the free receptor IL-2 in the subgroup B with increasing disease activity and in the subgroup with lupus nephritis. Statistically significant differences in values were identified between the subgroup A (non-active) and the subgroup LN (lupus nephritis) with p = 0.01 during the first set of the measurements. Fluctuation of sIL-2r levels in individual subgroups during the time of monitoring did not reach statistically important levels. In conclusion it could be said that potential practical utilization of the measurement of concentrations of the two mentioned molecules should be seen especially in monitoring disease activity because they don't contribute to SLE with needed information. Their always low values have favourable prognostic impact in monitoring patients with SLE and vise versa.

[Occurrence of celiac disease in siblings and offspring of patients with celiac disease]. / Výskyt celiakie u sourozencu a potomku nemocných s celiakií.

BACKGROUND: Incidence of the coeliac disease in our population is 0.3 to 0.5%, however, in direct relatives of patients with coeliac disease incidence rises up to 5-15%. Though the disease in not always clinically manifested, it can be identified by serologic screening. METHODS AND RESULTS: 96 patients treated for active coeliac disease and their 130 siblings were examined. In four of 130 siblings the disease clinically manifested and was confirmed with finding of enteropathy of jejunal mucosa. In 14 out of 126 asymptomatic siblings endomysial antibodies were identified and in 11 of them enteropathy was diagnosed. Coeliac disease was diagnosed in 15 (11.5%) persons from 130 siblings. Also 74 offsprings of 43 parent patients were examined. In four of them the coeliac disease manifested clinically and was later confirmed with finding of enteropathy. Antiglidin antibodies were tested in 45 asymptomatic offsprings with 6 positive results. Endomysial antibodies were tested in 25 asymptomatic offsprings with one positive result. Villous atrophy of jejunal mucosa was found in four asymptomatic offsprings. Coeliac disease was diagnosed in 8 (11.5%) persons from 74 offsprings. CONCLUSIONS: Coeliac disease represent a major risk for the patient's direct relatives. Goal-directed examination and the early diagnosis of the disease in childhood may help to prevent impairments in the development of the child and possible complications later.

[Idiopathic hypoparathyroidism with celiac disease--diagnostic and therapeutic problem]. / Idiopatická hypoparatyreóza s celiakií--diagnostické a lécebné problémy.

A middle-aged woman developed gradually diseases where most probably the common denominator is an autoimmune process. The authors describe the clinical condition persisting for seven years, where hypoparathyroidism was associated with coeliac disease which did not respond to dietary measures. Malabsorption was manifested by a selective block for calcium absorption from the gut. The impaired conversion of vitamin D2 to D3 is of interest; it has obviously a multifactorial etiology. The main problem is the patient's tendency to respond by tachyphylaxis to oral vitamin D administration. The only effective therapeutic procedure apart from those listed is so far parenteral administration of vitamin D3, calcitrol.

Effect of nitric oxide metabolism upon peripheral blood chemiluminiscence and leukocyte and erythrocyte adherence in multiple sclerosis.

BACKGROUND: It has become increasingly apparent that nitric oxide (NO) and related metabolic and effector systems play an important role in immune regulations, in inflammatory response and in cellular and tissue damage in multiple sclerosis (MS) and in its experimental models. OBJECTIVES: Various adherence interactions and metabolism of reactive oxygen species belong to the cellular functions associated with NO metabolism. The aim of the present study was to evaluate the effect of chemical compounds affecting NO metabolism on adhesive properties of leukocytes and erythrocytes and on the blood chemiluminiscence in MS patients in comparison with controls. The effects of NG-Methyl-L-Arginine (NGMLA), a specific competitive NO antagonist, L-arginin, NO precursor, and sodium nitroprusside, an NO-releasing agent were tested. METHODS: Luminol dependent chemiluminiscence was measured with and without respective drug administration, the results were expressed as chemiluminiscence index. Adherence of peripheral blood cells to nylon wool was used for the adherence studies, the results were expressed in a form of adhesivity index. Studies were performed in a group of patients with active multiple sclerosis, in a group of neurological controls and in a group of healthy controls. The differences between MS patients and control groups were statistically evaluated using Mann and Whitney U-test. MAIN RESULTS: Sodium nitroprusside stimulated the luminol-dependent chemiluminiscence in MS patients and inhibited it in both control groups. The difference between MS patients and the control patient subgroup was proved to be statistically significant. Statistically significant inhibition of leukocyte adherence by arginine was found in MS patients in comparison to healthy controls, in whom stimulation of adherence was observed. When differentiating the effect of the agents upon particular leukocyte subpopulations the only statistically significant difference was found with arginine and granulocytes, the trend being similar as in the total leukocyte adherence. Sodium nitroprusside further stimulated the adherence of erythrocytes in MS patients and inhibited it in controls. CONCLUSIONS: Results indicate nitric oxide-dependent alterations of oxidative metabolic burst and of cellular adherence properties in patients with active multiple sclerosis. Our findings also question possible participation of granulocytes and erythrocytes in multiple sclerosis pathogenetic cascade.

[The importance of determination of interleukin-10 in the blood of patients with systemic lupus erythematosus]. / Význam vysetrení interleukinu-10 v séru nemocných se systémovým lupus erythematodes.

BACKGROUND: Hitherto, not very numerous investigations provided so far only few often controversial findings on the importance of interleukin-10 (IL-10) in systemic lupus erythematosus (SLE). The objective of the present investigation was to assess whether there exist practically applicable relations between the serum level of IL-10, clinical and laboratory indicators of activity of the disease and serum levels of selected cytokines or their soluble receptors. METHODS AND RESULTS: The authors analyzed a group of 23 patients with SLE (23 women and 1 man, median age 37 years). Interleukin-10 and other cytokines were examined by the ELISA method, the clinical activity of the disease was evaluated by the ECLAM system (European Consensus Lupus Activity Measurement). Elevated IL-10 values (> 5 pg/ml) were assessed in 10 (43%) patients. Correlation analysis (Pearson's test, p < 0.05) revealed statistically significant relations between IL-10 levels and the activity of the disease, values of antibody levels against dsDNA and levels of the soluble receptor IL-2 (sIL-2R) in serum. Conversely, no relationship was revealed between values of IL-10 and values of C3 and C4 complement components, IL-1, IL-2, IL-6, sIL-6R, TNF-alpha, sTNFR-alpha and INF-gamma. CONCLUSIONS: Elevated IL-10 serum levels in patients with SLE did not have, with the exception of the index of clinical activity of the disease, antibodies against dsDNA and sIL-2R any statistically significant relations to laboratory indicators of disease activity and levels of selected cytokines and their soluble receptors.

[Vascular intercellular adhesive molecule-1 (VCAM-1)--a new indicator of activity in systemic lupus erythematosus?]. / Vaskulární intercelulární adhezivní molekula-1 (VCAM-1)--nový ukazatel aktivity systémového lupus erythematodes?

The objective of the submitted study was to evaluate the role of the serum level of the soluble form of the vascular adhesive molecule-1(VCAM-1) in systemic lupus erythematosus (SLE) and to evaluate the possible relation to selected clinical and laboratory indicators of activity of the disease and cytokine serum levels. The analyzed group comprised 20 women, median age 37 years (range 18-65 years). Elevated VACM-1 serum levels were detected in 18 subjects (90%). Statistical analysis (Pearson's test, p < 0.05) revealed a significant relationship between serum levels of VCAM-1 and the index of clinical activity of SLE evaluated by the ECLAM system (European Consensus Lupus Activity Measurement), values of the C3 complement component, the level of antibodies against dsDNA and serum levels of IL-10. No significant correlation was found with levels of the soluble receptor IL-2 (sIL-2R) and the C4 complement component in serum. From the investigation ensues that investigation of serum levels of the adhesive molecule of the VCAM-1 type is a promising method which makes early diagnosis of exacerbation of the disease possible. However, for evaluation of its real asset in clinical practice a more widely conceived longitudinal investigation is needed.

[The autoantibody profile and disease activity in patients with systemic lupus erythematosus]. / Autoprotilátkový profil a aktivita choroby u nemocných se systémovým lupus erythematodes.

Antinuclear antibodies are a group of autoantibodies which are typical for collagenous diseases. By means of the autoantibody profile different sub-groups of systemic lupus erythematosus (SLE) can be identified. This can serve as a certain prognostic factor of the affection. Patients with a negative antibody profile have fewer clinical and laboratory manifestations of SLE. Profile A (anti-dsDNA and/or anti-Sm has, as compared with patients with a negative antibody profile, more frequent organ manifestations. Patients with profile B (anti-RNP) have a higher frequency of Raynaud's phenomenon. Profile C (anti-Ro, anti-La) is characterized in particular by photosensitivity of the skin and secondary Sjögren's syndrome. Profile D (antibodies against centromeres and/or Scl-70) are found in subjects with SLE with traits of scleroderma. Finally profile E (antibodies against histones) are found in SLE induced by drugs. In the submitted study in 28 patients with SLE autoantibodies anti-dsDNA, anti-DNP, extracted nuclear antibodies (ENA-Sm,Ro,La, histones, Sm/RNP, Scl-70) were evaluated and different subgroups of SLE were assessed. Attention was paid to their common characteristics and the activity of the disease. Associations of clinical activity of the disease expressed by the ECLAM index (European Consensus Lupus Activity Measurement) were tested as well as anti-dsDNA levels and also the association of the disease activity with C3 and C4 constituents of complement, CRP and circulating immunocomplexes in serum. Positivity of the antinuclear factor (ANF) was found in 21 patients, while in 7 subjects who were in clinical and laboratory remission, ANF was negative. A negative antibody profile was recorded in 9 patients, profile A was found in 13, 1 patient had profile B, and 4 patients had profile C. Antibody profile D was not found in the group. When using regression analysis and Pearson s correlation coefficient, correlations were found between anti-dsDNA values and the system ECLAM (r = 0.72, p < 0.01), anti-dsDNA and C3 levels (r = -0.59, p < 0.01), C4 (r = -0.50,, p < 0.01), and between the ECLAM system and C3 (r = -0.60, p 0.01) and C4 (r = -0.52, p < 0.01) and also between C3 and C4 mutually (r = 0.72, p < 0.01). From the submitted investigation ensues that investigation of antinuclear antibody levels in SLE is important not only for assessment of the diagnosis of the disease and its activity but also for assessment of the subgroups of the disease and for prediction of its development. As to other indicators of activity, assessment of the C3 and C4 constituents of complement is still important.

The level of neopterin in blood serum as one of the possible prognostic markers of Crohn's disease activity.

Neopterine is considered to be one of the markers of immunity system activity. An increased level of this pteridine derivate is determined in blood or urine in infections, transplant rejections, and in other conditions accompanied by changes in the immunity system. Monitoring its level in various body liquids can be important when assessing the immunity system condition, nevertheless, its non-specificity causes difficulties when interpreting the results. Up to now, very few papers have been published dealing with the determination of neopterine levels in patients with IBD. We studied a group of 25 patients who are monitored in a longterm manner at our clinic suffering from Crohn's disease. During 1993 and 1994, we were determining their levels of neopterine in blood together with other parameters. It was a group of patients chosen at random in different stages of disease, and we were interested in the correlation of the height of neopterine level with the activity of Crohn's disease. We proved statistically unimportant correlation between the height of neopterine level in blood serum and Crohn's disease activity. The question is discussed whether the examination of this marker be useful when monitoring Crohn's disease activities.

C-ANCA and p-ANCA in patients with Crohn's disease.

In 32 patients suffering from various stages of activity of Crohn's disease, c-ANCA and p-ANCA in serum were examined. Altogether, 54 blood samples were examined. In 21 patients of 32, i.e. 65.6%, at least in one examination of c-ANCA in serum the value was higher than 9 units (positive finding). In 10 patients of 32, i.e. 32.2%, at least in one examination of p-ANCA in serum the value was higher than 9 units (positive finding). In the case of high activity of Crohn's disease, c-ANCA in serum were higher than 9 units (positive finding) in 69.6% examinations, in case of low activity, values of c-ANCA in serum were higher than 9 units (positive finding) only in 9.5% examinations. p-ANCA in serum were, when the activity was high, positive in 46.1% examinations, while when the activity of Crohn's disease was low, p-ANCA were positive in 9.5%. The question of specificity of findings is discussed.

Immunological profile of patients with recurrent respiratory infections.

The authors present an analysis of the results of laboratory immunological examination of 52 patients suffering from recurrent respiratory infections. Besides the typical laboratory correlates of chronic inflammation, several findings indicated the depressed function of cellular and partially humoral immunity. The immunological parameters, most frequently decreased in comparison with normal values, were as follows: the response to the recall antigens of Imunoskintest (lower in 54% of patients), the relative number of CD3+ lymphocytes (35%), the CD4/CD8 ratio (37%), phagocytic activity (37%) and also serum IgA (12%). This means that more than two thirds of patients displayed at the time of examination a substantial alteration of one or more immunological parameters, the depression of cellular immunity was much more pronounced. It is concluded that the laboratory immunological examination of patients with recurrent respiratory infections is very important for revealing of an underlying cause of the disease and for indicating the adequate immunomodulatory treatment.