Increased frequency of CD16+ monocytes and the presence of activated dendritic cells in salivary glands in primary Sjögren syndrome.

OBJECTIVES: In the salivary glands of patients with primary Sjögren Syndrome (pSjS) an accumulation of dendritic cells (DCs) is seen, which is thought to play a role in stimulating local inflammation. Aberrancies in subsets of monocytes, generally considered the blood precursors for DCs, may play a role in this accumulation of DCs. This study is aimed at determining the level of mature CD14lowCD16+ monocytes in pSjS and their contribution to the accumulation of DCs in pSjS. METHODS: Levels of mature and immature monocytes in patients with pSjS (n = 19) and controls (n = 15) were analysed by flow cytometry. The reverse transmigration system was used for generation of DCs generated from monocyte subsets. The phenotype of DCs in pSjS salivary glands was analysed using immunohistochemistry. In vivo tracking of monocyte subsets was performed in a mouse model. RESULTS: Increased levels of mature CD14lowCD16+ monocytes were found in patients with pSjS (mean (SD) 14.5 (5.5)% vs 11.4 (3.4)%). These cells showed normal expression of chemokine receptor and adhesion molecules. Mature monocytes partly developed into DC-lysosome-associated membrane glycoprotein (LAMP)+ (19.6 (7.5)%) and CD83+ (16 (9)%) DCs, markers also expressed by DCs in pSjS salivary glands. Monocyte tracking in the non-obese diabetic (NOD) mouse showed that the homologue population of mature mouse monocytes migrated to the salivary glands, and preferentially developed into CD11c+ DCs in vivo. CONCLUSIONS: Mature monocytes are increased in pSjS and patient and mouse data support a model where this mature monocyte subset migrates to the salivary glands and develops into DCs.

Serological markers to differentiate between ulcerative colitis and Crohn's disease.

AIM: To assess prospectively the value of three serological tests for differentiating between ulcerative colitis and Crohn's disease, used either alone or combined. METHODS: Coded serum samples from 63 patients with ulcerative colitis and 67 patients with Crohn's disease were analysed. Detection assays for the presence of perinuclear antineutrophil cytoplasmic antibodies (pANCA), serum agglutinating antibodies to anaerobic coccoid rods, and specific IgG antibodies against a Kd-45/48 immunological crossreactive mycobacterial antigen complex (ImCrAC) were studied. Sensitivity, specificity, pre- and post-test probabilities, likelihood ratios, and predictive values of each of these serological tests were determined. RESULTS: The sensitivity and specificity of the pANCA test for the diagnosis of ulcerative colitis were 61 and 79%, respectively. The serum agglutination test for anaerobic coccoid rods had a sensitivity of 42% and a specificity of 89% for a diagnosis of Crohn's disease. The sensitivity of specific IgG antibodies against Kd-45/48 ImCrAC in diagnosing Crohn's disease was 70% and specificity 60%. Although 100% specificity was achieved by combining all three tests in a small group of patients with Crohn's disease (n = 20), combining two or more tests had no additive clinical value. No correlation was found between the presence of any one of these antibodies and disease activity, duration, or localisation of disease. Surgery or medical treatment did not influence the presence of antibodies or the antibody titre. CONCLUSIONS: The value of these tests in the differential diagnosis between ulcerative colitis and Crohn's disease is limited, but the high predictive values and specificities of different tests for both diseases suggest that these tests may be of help in studying disease heterogeneity and in defining different subgroups of patients with different pathogenesis.

Retinal vasculitis occurring with common variable immunodeficiency syndrome.

PURPOSE: To report severe retinal vasculitis causing decreased vision in three patients with the common variable immunodeficiency syndrome. METHOD: Case report. Three patients with common variable immunodeficiency syndrome developed decreased vision secondary to retinal vasculitis. Fluorescein angiography was performed in all three patients. Peribulbar injections were given in one patient, and two patients were treated with oral steroids and cyclosporin. RESULTS: All three patients were young and had classic common variable immunodeficiency syndrome. Bilateral retinal vasculitis and diffuse retinal edema were present in all three patients, and two patients had retinal neovascularization in the absence of ischemia. No evidence of intraocular infection was present, and none was detected systematically. Visual acuity decreased in five of the six eyes and was responsive to treatment in only one patient (both eyes). CONCLUSION: Retinal vasculitis may be another autoimmune manifestation of common variable immunodeficiency syndrome.

Hemiparkinsonism in a patient with primary Sjögren's syndrome. A case report and a review of the literature.

A 55-year-old woman presented with hemiparkinsonism on the left side. Clinical features suggested primary Sjögren's syndrome. Eye tests and a salivary gland biopsy confirmed this diagnosis. Magnetic resonance imaging of the brain showed distinct linear lesions of increased intensity on the T2-weighted images in the right striatum and globus pallidum. Treatment with prednisone and azathioprine did not give any improvement of the neurological symptoms. To date, hemiparkinsonism in association with primary Sjögren's syndrome has not been reported.

CD5+ B lymphocytes and other lymphocyte subsets in primary Sjögren's syndrome.

CD5+ B cells and other lymphocyte subsets were analyzed by flow cytometry in patients with primary Sjögren's syndrome (pSS), in healthy subjects (HS) and in patients with various control diseases. When compared with HS, patients with pSS were found to have similar levels of CD5+ B cells and decreased levels of CD8+ T cells (P = 0.0003). When compared with patients with various other diseases, however, the number of CD5+ B cells in pSS was more than twice as high (P = 0.0002), whereas no difference was found between numbers of CD8+ T cells. When the number of CD5+ B cells was expressed as a percentage of total B cells, the results obtained were similar to those with absolute numbers. Determination of lymphocyte subsets may be used as a diagnostic aid for Sjögren's syndrome in selected patients with suspected immunological diseases of unknown type.

Agglutination of Eubacterium and Peptostreptococcus species as a diagnostic test for Crohn's disease.

The anaerobic fecal flora of patients with Crohn's disease has been found to differ from the flora of healthy subjects by higher numbers of anaerobic gram-negative rods and gram-positive coccoid rods. Agglutinating antibodies against strains of coccoid rods, belonging to species of Eubacterium and Peptostreptococcus were detected in much higher percentages of sera from patients with Crohn's disease than in those from patients with ulcerative colitis, and healthy subjects. On the basis of the results of agglutination reactions with four strains, the probability of Crohn's disease can be estimated. In this paper, the background of the test ist described and its use and limitations are reviewed.

Plasma lysozyme levels and decay of neutrophilic granulocytes in patients with Crohn's disease.

In patients with Crohn's disease, the lysozyme concentration in plasma was compared with the Crohn's disease activity index, the granulocyte lysozyme content, the number of circulating neutrophilic granulocytes in peripheral blood and the unsaturated vitamin B12-binding capacity of the plasma cobalophilins as an index for the granulocyte decay. There was no difference in the lysozyme content of granulocytes from patients and controls. Patients with increased plasma lysozyme levels suffered from a more active disease and had larger numbers of circulating granulocytes in their blood than those with normal plasma lysozyme levels. The plasma lysozyme levels correlated well with the cobalophilin levels, in patients with increased levels even more markedly than in those with normal levels. The results suggest that in patients with Crohn's disease increased plasma lysozyme levels are due to an increase in granulocyte decay.

Cost analysis of regular and filgrastim treatment in patients with refractory chronic rhinosinusitis.

In this double blind randomized placebo controlled clinical trial of filgrastim in chronic sinusitus, we analyzed costs of a 24-week interval in which filgrastim was administered. Since we hypothesized that the scheduled preventive visits within the trial might cause savings as compared to the regular situation in which these patients have a strong tendency to visit the outpatient clinic immediately in case of complications, direct medical costs within the trial were also compared to costs of regular treatment. The difference in costs between both trial groups was driven by the filgrastim costs (Euro 5108). If filgrastim costs were left out of consideration, no significant difference in direct medical costs remained between the filgrastim and placebo groups (Euro 2904 and Euro 2765, respectively). Indirect medical costs also showed no significant differences. Within a regular situation, costs of a 24-week interval were Euro 896. As filgrastim treatment had already been shown not to improve the quality of life, its cost-effectiveness in these patients can only be favourable in case of major clinical improvements. Furthermore, scheduled preventive visits in chronic sinusitis patients appear not to cause savings as compared to the situation in which patients are only seen in case of recurrences.

Differentiation between diseases using a programmable hand-held calculator and Bayes' theorem: application to lower gastrointestinal tract disorders.

A program is presented for the Hewlett-Packard HP-41C programmable pocket calculator that computes posterior probabilities of more than 200 diseases on the basis of Bayes' theorem. Data for specific applications are stored in ASCII files. The program and data are retained in the non-volatile memory of the calculator. An example of the application of the program to six lower gastrointestinal tract disorders is given using data from the literature.

Interpretation of laboratory tests using a programmable hand-held calculator: calculation of posterior probability and relative risk on the basis of prior probability and sensitivity, specificity and result of the test.

A program is presented for the Hewlett-Packard HP- 41C programmable hand-held calculator that calculates the posterior probability and relative risk of an event (e.g. a disease) on the basis of prior probability (prevalence) and sensitivity, specificity and result of the test.

Differentiation between two diseases using a programmable hand-held calculator and Bayes' theorem: application to Crohn's disease and ulcerative colitis.

A short general program is presented for the Hewlett-Packard HP-41C programmable pocket calculator that calculates posterior probabilities of two diseases on the basis of frequencies of nosographic characteristics using Bayes' theorem. Extended memory and functions modules allow the storage of many data in an ASCII file.

iTRAQ-based proteomic profiling of the marine medaka (Oryzias melastigma) gonad exposed to BDE-47.

A recent study demonstrated that 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) may have an adverse effect on the reproduction in marine medaka (Oryzias melastigma), but the molecular mechanisms remain largely unknown. In this study, we investigated the protein expression profiles of male and female gonads of O. melastigma exposed to dietary BDE-47 at two dosages (0.65 and 1.30 µg/g/day, respectively) for 21 days. Extracted proteins were labeled with iTRAQ and analyzed on a MALDI TOF/TOF analyzer, as results, 133 and 144 unique proteins were identified in testis and ovary, respective, and they exerted dose- and sex-dependent expression patterns. In testis, among the 42 differentially expressed proteins; down-regulation of histone variants and parvalbumins implicated BDE-47 may disrupt the spermatogenesis and induce sterility in fishes. In ovary, 38 proteins were differentially expressed; the elevation of vitellogenins and apolipoprotein A-I expression indicated BDE-47 acts as an estrogen-mimicking compound and led to reproductive impairment in O. melastigma.

Gender-specific transcriptional profiling of marine medaka (Oryzias melastigma) liver upon BDE-47 exposure.

Marine medaka (Oryzias melastigma) were exposed to 2,2',4,4'-tetrabromodiphenyl ether (BDE-47) to investigate the gender-specific transcriptional profiles of liver tissue in response to this flame retardant. A cDNA library of O. melastigma was constructed, and 2304 clones were amplified from the library to fabricate a cDNA microarray. Sequences of these genes were assembled into 1800 sequences using Geneious, a bioinformatics software. Corresponding expressed sequence tags were blasted against the National Centre for Biotechnology Information non-redundant database and further classified into various biological categories according to the Gene Ontology project. Male and female three-month-old were fed a diet of BDE-47 contaminated Artemia at low dosage (290.3±172.3ng BDE-47/day) and high dosage (580.5±344.6ng BDE-47/day) for 5 and 21 days, respectively. The transcriptional profiles of O. melastigma liver were then generated by the species-specific cDNA microrarray. The results from microarray analysis suggested very different gene expression patterns between males and females for both BDE-47 exposure-dose and exposure-time, with male livers having stronger gene regulatory responses than female livers. Importantly, our results revealed that in male O. melastigma only, BDE-47 exposure may activate phosphoinositide-3-kinase and mitogen-activated protein kinase, proteins that play importance roles in cell growth, proliferation and survival.

The human faecal flora and Crohn's disease.

The aetiology of Crohn's disease (CD), a chronic inflammatory disease of the intestines, is not known but a genetic predisposition to CD has been well demonstrated. Studies on the isolation of bacteria from tissues and the faecal flora from CD patients are reviewed and their results compared with those from our laboratory. Serum antibodies against a variety of bacteria have been described but none has been found to be specific for CD. In our laboratory, higher numbers of coccoid anaerobes have been found in the faecal flora of CD patients. The use of agglutination reactions with four coccoid anaerobes as a diagnostic aid for CD is reviewed. No evidence has been found, however, for a primary role of the agglutinating antibodies in the pathogenesis of CD. On the other hand, one of the coccoid strains has been found to have several characteristics suggesting a role in CD, such as activation of the alternative pathway of complement, the failure of opsonization by specific IgG antibody and the binding of (non-specific) IgG onto the bacteria by the Fc portion. The composition of the faecal flora has been found to depend on genetic characteristics of the host. It is suggested that the genetic predisposition to CD is based on the genetically determined indigenous faecal flora present.

A possible role of Eubacterium and Peptostreptococcus species in the aetiology of Crohn's disease.

The faecal flora of patients with Crohn's disease has been found to contain higher numbers of anaerobic gram-negative rods and gram-positive coccoid rods than the flora of healthy subjects. In a considerable percentage of sera from patients with Crohn's disease, agglutinating antibodies were found against four strains of the coccoid rods, in contrast to sera from patients with ulcerative colitis, other diseases and healthy subjects. In the present study, coccoid strains C18, Me46 and Me47 were found to activate complement by the alternative pathway. IgG antibodies to strains Me44, C18 and Me47 had opsonic properties but those to strain Me46 failed to induce phagocytosis. The data suggest an inadequate immune defence against strain Me46 in patients with Crohn's disease. The possible role of strain Me46 in the pathogenesis of Crohn's disease is discussed.

Serum agglutinins to Eubacterium and Peptostreptococcus species in Crohn's and other diseases.

Sera from patients suffering from Crohn's and other diseases and from healthy subjects were tested for agglutinins to anaerobic, gram-positive coccoid rods belonging to species of Eubacterium and Peptostreptococcus. Four strains labelled Eubacterium contortum (two strains), Eubacterium rectale and Peptostreptococcus productus were agglutinated by a higher percentage of sera from patients with Crohn's disease than from healthy subjects and from patients with liver and intestinal diseases (including ulcerative colitis), ankylosing spondylitis, granulomatous diseases, diseases of immunity and malignancies. The agglutinins were of the IgG and IgM classes and strain-specific; the titres were low. The results obtained with sera from patients with Crohn's disease and healthy people were subjected to discriminant analysis to estimate the probability, based on the combined results with the four strains, that a patient suffers from Crohn's disease. When sera giving an a posteriori probability greater than or equal to 0.95 (a priori probability = 0.5) were considered positive, the test with four strains had a sensitivity of 54% and a specificity of nearly 100%. The results with sera submitted for diagnosis showed that positive reactions in patients with a diagnosis apparently incompatible with Crohn's disease were within acceptable limits.

Binding of Coprococcus comes to the Fc portion of IgG. A possible role in the pathogenesis of Crohn's disease?

Previous studies have shown that the fecal flora of patients with Crohn's disease (CD) differed from the flora of healthy subjects by a higher number of anaerobic gram-positive coccoid rods. Sera from patients with CD agglutinated four strains of coccoid rods (Me44, C18, Me46 and Me47) more frequently and stronger than sera from healthy subjects and patients with other diseases. One of these bacteria, Coprococcus comes strain Me46, was not ingested by neutrophils after coating with specific IgG. In the present study, therefore, the binding of IgG and of Fab and Fc fragments to the four coccoid rods was investigated using immunofluorescence and absorption techniques. Results were compared with those obtained with Staphylococcus aureus strain Cowan I and showed that Me46, like S. aureus, bound (nonspecifically) IgG through its Fc portion whereas strain Me47 bound IgG through the Fab portion. Possible implications of the findings for CD are discussed.