Low circulating Dickkopf-1 and its link with severity of spinal involvement in diffuse idiopathic skeletal hyperostosis.

OBJECTIVE: Dickkopf-1 (DKK-1) is an inhibitor of osteoblastogenesis, and its lower levels are linked to new bone formation. The aim of this study was therefore to explore serum levels of DKK-1 and to evaluate DKK-1's association with the severity of spinal involvement in diffuse idiopathic skeletal hyperostosis (DISH). METHODS: Serum levels of total and functional DKK-1 and C-reactive protein (CRP) were measured in 37 patients with DISH and 22 healthy age and sex-matched controls. Plain radiographs of the cervical and thoracic spine were performed, and the diagnosis of DISH was defined using the Resnick criteria. Patients were divided into three groups based on spinal involvement. Bone mineral density (BMD) and bone turnover markers were evaluated in patients with DISH. RESULTS: The levels of total serum DKK-1 were significantly lower in patients with DISH than in healthy controls (p<0.0001). Importantly, low serum levels of DKK-1 were associated with more severe spinal involvement in DISH, independent of age, sex, disease duration, CRP, bone turnover markers or BMD. However, these findings were less significant for functional DKK-1. CONCLUSION: These observations indicate that DKK-1 may play a significant role in bone formation during DISH.

No evidence for linkage between the hereditary angiooedema clinical phenotype and the BDKR1, BDKR2, ACE or MBL2 gene.

Hereditary angiooedema (HAE) is a life-threatening disease with poor clinical phenotype correlation with its causal mutation in the C1 inhibitor (SERPING1) gene. It is characterized by substantial symptom variability even in affected members of the same family. Therefore, it is likely that genetic factors outside the SERPING1 gene have an influence on disease manifestation. In this study, functional polymorphisms in genes with a possible disease-modifying effect, B1 and B2 bradykinin receptors (BDKR1, BDKR2), angiotensin-converting enzyme (ACE) and mannose-binding lectin (MBL2), were analysed in 36 unrelated HAE patients. The same analysis was carried out in 69 HAE patients regardless of their familial relationship. No significant influence of the studied polymorphisms in the BDKR1, BDKR2, ACE and MBL2 genes on overall disease severity, localization and severity of particular attacks, frequency of oedema episodes or age of disease onset was detected in either group of patients. Other genetic and/or environmental factors should be considered to be responsible for HAE clinical variability in Caucasians.

Increased serum levels of B cell activating factor (BAFF) in subsets of patients with idiopathic inflammatory myopathies.

OBJECTIVE: To investigate serum levels of B cell activating factor (BAFF) in patients with myositis and correlate these to autoantibody profile, clinical phenotype and treatment. METHODS: BAFF levels in sera from 49 patients with dermatomyositis, 44 with polymyositis, 6 with inclusion body myositis and 30 matched controls were measured by ELISA. Specific autoantibodies were detected by line blot and western blot assays. RESULTS: Serum levels of BAFF were significantly higher in patients compared to healthy controls (p = 0.003). Patients with anti-Jo-1 autoantibodies had higher BAFF levels than control individuals (p<0.003) or patients without any specific autoantibodies (p<0.05). Patients with dermatomyositis had higher BAFF levels compared to polymyositis (p<0.05). Patients with interstitial lung disease (ILD) had higher BAFF levels than patients without ILD (p<0.05) or controls (p<0.01) but this could be explained by presence of anti-Jo-1 autoantibodies. BAFF levels correlated with serum creatine kinase (CK) (rs = 0.365, p = 0.0005) but not with C-reactive protein (CRP) levels. A negative correlation of BAFF levels with glucocorticoid daily dose for all patients (rs = -0.292, p = 0.003) and with cumulative glucocorticoid doses in early myositis cases (rs = -0.659, p<0.001) was recorded. CONCLUSION: Our finding of elevated serum levels of BAFF in patients with myositis with described phenotypes together with the correlations between levels of BAFF and CK and a negative correlation with dose of glucocorticoids, indicate that BAFF could be a potential therapeutic target in such cases.

Endogenous HLA-DR-restricted presentation of the cartilage antigens human cartilage gp-39 and melanoma inhibitory activity in the inflamed rheumatoid joint.

OBJECTIVE: The cartilage proteins melanoma inhibitory activity (MIA) and human cartilage gp-39 (HC gp-39) are candidate autoantigens in rheumatoid arthritis (RA). The present study was undertaken to investigate the endogenous HLA-DR4-restricted presentation of these self proteins, in order to seek in vivo evidence in support of their potential immunologic role. METHODS: MIA and HC gp-39 were assessed in synovial fluid (SF) by enzyme-linked immunosorbent assay and in synovial tissue (ST) by immunohistochemistry. Presentation by SF cells was investigated using specific, HLA-DR-restricted T cell hybridomas. RESULTS: MIA and HC gp-39 were detected in RA SF and ST, as well as in specimens from patients with other forms of arthritis. When HC gp-39-specific and MIA-specific HLA-DR4-restricted T cell hybridomas raised in HLA-DR4-transgenic mice were incubated with RA SF cells as antigen-presenting cells in the presence of HC gp-39 or MIA peptides, the corresponding T cell hybridomas showed strong responses, which were blocked by anti-HLA-DR antibodies. Weaker but qualitatively similar responses were observed with exogenous protein, indicating uptake and processing of these antigens by SF cells. More importantly, without addition of peptide or protein, endogenous presentation of MIA and HC gp-39 was detected in SF cells from 53% and 80% of HLA-DRB1*0401-positive RA patients, respectively. In addition, SF cells from 3 of 10 patients with spondylarthritis exhibited endogenous HC gp-39 presentation. CONCLUSION: These data indicate that immunodominant epitopes of MIA and HC gp-39 are actively presented in an HLA-DR-restricted manner in the inflamed RA joint. The question remains as to whether this leads to activation of autoreactive T cells, which could play a role in either the immunopathology or the immunomodulation of arthritis.

Periprosthetic osteolysis and its association with RANKL expression.

Extensive osteolysis adjacent to orthopedic implants is often associated with wear particles of prosthetic material. The activation of the RANKL/RANK/OPG system is considered to be a likely cause of periprosthetic osteolysis leading to implant failure. The aim of this study was to examine the possible correlation between the clinical extent of osteolysis, the number of wear particles and expression of the osteoclastic mediator RANKL (receptor activator of nuclear factor kappa B ligand) in the tissues around aseptically loosened cemented and non-cemented total hip replacements. Periprosthetic tissues were harvested from 59 patients undergoing revision of hip replacement for aseptic loosening. We observed RANKL-positive cells in 23 of our 59 patients, their presence was noted predominantly in tissues with a loosened cemented endoprosthesis. We have found that RANKL is present only in tissues with a large amount of wear debris and predominantly in cases involving loosened cemented implants.

[Prevalence of hepatitis G virus infection (HGV) in intravenous immunoglobulin recipients in the Czech Republic]. / Výskyt infekce virem hepatitidy G (HGV) u pacientu lécených intravenózními imunoglobuliny v Ceské republice.

The prevalence of hepatitis G virus (HGV) in the serum of intravenous immunoglobulin (IVIG) recipients was studied and risk related to HGV positivity was considered. Although its pathogenicity is unclear, HGV is likely to cause liver disease or lymphoproliferation. Twenty (23%) of 86 tested MG patients were HGV RNA positive. Of the HGV positive patients, three (15%) showed mild elevation of liver enzymes and one (5%) was diagnosed with chronic lymphatic leukaemia prior to the institution of MG replacement. It can be concluded that the HGV prevalence among IVIG recipients is high but is not associated with signs of either liver disease or lymphoproliferation.

[Risk factors for ischemic heart disease in patients with chronic fatigue syndrome]. / Rizikové faktory ischemické choroby srdecní u pacientu s chronickým únavovým syndromem.

Risk factors of coronary artery disease (CAD) between a group of patients suffering of chronic fatigue syndrome (CFS) and a control group of healthy persons (whose exercise activity was not health-limited) were compared. Thirty three patients (27 women, 6 men, average age 39.9 +/- 11.7 years) and the same number of controls matched in age (39.8 +/- 10.3 years), gender and body weight. The Minnesota Questionnaire (by Taylor) and the Compendium of Physical Activities (by Ainsworth) were used to estimate total energetic expenditure in exercise activity as well as in job. The risk factors of CAD in the patients with CFS were not higher than in the control group. Aerobic physical fitness, basic anthropometric data, blood pressure, spectrum of blood lipoproteins, blood uric acid and smoking habits were not different between the compared groups. Patients suffering from CFS had lower total energetic expenditure in exercise activity. Nevertheless, this significant difference in sports activity was not large enough to cause any difference in risk factors of CAD between the CFS patients and the control group.

[CD5-positive B lymphocytes and antinuclear antibodies]. / CD5 pozitivní B lymfocyty a antinukleární protilátky.

BACKGROUND: The production of natural autoantibodies incl. antinuclear antibodies (ANA) is ascribed to lymphocytes which have a CD5 molecule on their surface. The role of CD5 positive B lymphocytes in the induction of autoimmunity is obscure so far. The authors focused their attention on the incidence of antinuclear antibodies (AA) in subjects with different diseases and sought a relationship with the ratio of CD5 positive B lymphocytes in the peripheral blood stream. METHODS AND RESULTS: CD5 positive lymphocytes were assessed on a flow cytometer using monoclonal anti CD5 and anti CD19 antibodies. Antinuclear antibodies are detected by indirect fluorescence on a substrate of human leucocytes and HEP-2 cells. In a group of 134 subjects the authors did not provide evidence of a direct relationship between the relative number of CD5 positive B lymphocytes in the peripheral blood stream and the presence of ANA (IgG, IgA, IgM), not even in 33 patients with autoimmune diseases. In 86 patients the authors revealed that antinuclear antibodies type IgM predominate in patients with repeated respiratory infections. In systemic diseases the isotype IgG predominates (p = 0.01). After immunosuppressive treatment with a favourable clinical effect the ANA isotype IgG disappears and isotype IgM is found more frequently. The incidence of the ANA isotype IgM is significantly higher in healthy subjects aged over 60 years than in younger subjects (12.5%/6.1%, p = 0.06), and more frequent in women (p = 0.05). CONCLUSIONS: The presence of antinuclear antibodies is not associated with the amount of CD5 positive B lymphocyte in the peripheral blood stream.

[Diagnostic and pathogenic significance of antineutrophil cytoplasmic antibodies in vasculitides]. / Diagnostický a patogenetický význam protilátek proti cytoplazmatickým antigenum neutrofilu vaskulitid.

OBJECTIVES: The study represents a complex report on diagnostic and pathogenic significance of antineutrophil cytoplasmic autoantibodies (ANCA). Our report is founded on the results of our clinical and experimental studies. METHODS: ANCA was examined by indirect immunofluorescence, or ELISA-anti MPO. The data for evaluation of diagnostic ANCA associations were gained by means of questionnaires. The experimental part involved examination of the impact of sera containing ANCA and monoclonal antibodies (MoAb) against proteinase 3 and control MoaB specific for neutrophils. RESULTS: Within the course of 1 year we have examined 1010 samples, ANCA was found in 65 patients: 26 cases of systemic vasculitis, 12 cases of other autoimmune diseases, 9 cases of isolated glomerulonephritis, 7 cases of inflammatory intestinal diseases, 6 cases of ophthalmic inflammatory diseases, and 6 cases of other diseases. Vasculitis of small vessels found in our group of patients are concomitted by ANCA positivity in 90% (Wegener's granulomatosis 15/16, Churg--Strauss's syndrome 3/5, MPA 9/9), vasculitis of medium vessels 12% (classical polyarteritis nodosa 1/2, Kawasaki disease 0/2, Henoch--Schönlein's purpura 0/4, Takayasu's arteritis 1/4). The ANCA-containing sera inhibit the candidacidal activity by 65% in average. THe inhibitory impact is yielded by anti-proteinase MoAb, in contrast to the control MoAb. CONCLUSION: c-ANCA in high concentration is specific for active form of Wegener's granulomatosis, other types of ANCA in low titres are not of diagnostic value for vasculitis. ANCA is concomitted especially by small vessels vasculitis. The proved inhibition of microbicidal PMN activity by ANCA can participate in some clinical signs of immunodeficiencies in vasculitis. (Tab. 5, Ref. 18.).

[The spectrum of diseases associated with antineutrophil cytoplasmic antibodies]. / Spektrum chorob sdruzených s výskytem protilátek proti cytoplazme neutrofilu.

BACKGROUND: Antineutrophil cytoplasmic antibodies (ANCA) are considered as a diagnostic marker of systemic vasculitis and rapidly progressing glomerulonephritis. They are frequently associated with other pathological conditions. The author's objective was to analyze a group of patients examined at the Department of Clinical Immunology of the First Medical Faculty Charles University in the course of one year and evaluation of the diagnostic asset of the ANCA examination. METHODS AND RESULTS: ANCA was assessed in serum using the method of indirect immunofluorescence. In cases of p-ANCA the ELISA test was used to assess antimyeloperoxidase antibodies. Clinical data were evaluated on the basis of analysis of a questionnaire sent to doctors of patients where ANCA was detected. In 1992 1010 specimens were examined and in 115 ANCA was detected. This was the case in a group of 65 patients with the following diagnoses: systemic vasculitis 26x, systemic immunopathological diseases 12x, renal diseases 9x, inflammatory bowel diseases 7x, inflammatory eye diseases 6x, monoclonal paraproteinaemia 2x, other 4x. The patients had renal affections (45%), affections of the joints (20%), skin (18%), eyes (14%), lungs (12%), GIT (11%), ENT (9%), and nerves (9%). Type c in titres above 1:160 was always associated with the diagnosis of Wegener's granulomotosis, in other instances c- or x-types titres of 1:20 were involved. Antibodies against myeloperoxidase were detected in four instances. CONCLUSIONS: The authors confirmed that c-ANCA in high concentrations are specific for the predominantly active form of Wegener's granulomatosis. The group of patients with p-ANCA is too small to generalize the diagnostic impact. Low ANCA concentrations are found in many other diseases. The importance of ANCA can be evaluated only in a clinical context and the importance of their presence must not be overestimated.

[Essential mixed cryoglobulinemia and the kidneys]. / Esenciální smísené kryoglobulinémie a ledviny.

The authors describe their experience with the treatment of nine patients where "essential" mixed cryoglobulinaemia was diagnosed. In addition to extrarenal symptoms all suffered from proliferative glomerulonephritis, most frequently mesangiocapillary. The latter was manifested in most instances during the first examination by nephrotic syndrome and reduced glomerular filtration. Two patients had only asymptomatic proteinuria with erythrocyturia. The majority suffered from arterial hypertension. The patients were treated with prednisone and cyclophosphamide, using initial doses after which the extrarenal symptoms receded promptly. Regression of proteinuria and normalization of glomerular filtration developed more slowly. Only in one patient the glomerulonephritis had a progressive course and terminated by chronic renal failure. One patient died from septicaemia and one from cardiac failure. The authors assume that early diagnosis and effective combined immunosuppression can ensure a favourable course of this rare disease.

[Personal experience with detection of antibodies against neutrophil cytoplasm]. / Nase zkusenosti s vysetrováním protilátek proti cytoplazme neutrofilu (ANCA).

Detection of antibodies against components of neutrophil cytoplasm (ANCA) is of diagnostic value in Wegener's granulomatosis and in some other types of vasculitis. In the submitted paper the authors discuss methodical aspects of ANCA detection. The most frequently used method for assessment of ANCA is indirect immunofluorescence, at present it is possible to use also the ELISA method. Based on their own group of patients the authors discuss the association of ANCA with various types of diseases and the importance of assessing ANCA during monitoring of immunosuppressive treatment.

Cell-mediated response to BCG investigated by leukocyte migration test from an agarose droplet.

The modified leukocyte migration test (LMT) from an agarose droplet with the antigen stimulation (by BCG) is proposed in the present work. The BCG concentrations ranging from 1.25 up to 130 mg/l were used to examine 20 tuberculin (PPD) skin test negative and 10 PPD positive patients, which suffered from lung diseases. The optimal concentrations were 6.25 and 25.0 mg/l. The mean index values of a 20 membered control group ranged from 0.7 up to 0.88 when stimulated with the lower BCG concentration, and they were of 0.53 up to 0.69 at higher BCG concentration. In spite of its suitable indicatory properties as to ascertain cell mediated immunity state, the LMT with the BCG were of no use as a tuberculin skin test correlate.