bluuebunny/crossref_metadata_2025 huggingface dataset:
Binding of human C-reactive protein to monocytes: analysis by flow cytometry
Authors: BALLOU S P , CLEVELAND R P
Published: July 2010

An opsonic role has been proposed as a major function of C-reactive protein (CRP) in humans. In support of this hypothesis, recent radiolabcltcd ligand binding studies have provided evidence for the presence of specific receptors for soluble human CRP on human phagocytic cells, including ncutrophits and monocytes. In order to confirm specific binding of CRP to monocytes and to quantify the percentage of such cells capable of expressing binding sites, we employed a sensitive biotin-avidin fluorescence assay to study the CRP-monocyte interaction. It was observed that 67% of monocytes bound biotinylated CRP in a dose-dependent manner, that the binding was calcium dependent, and that it could be inhibited by 60% in the presence of a greater than 20-fold excess of competing native CRP. In other experiments, neither IgG nor heat-aggregated IgG inhibited the binding of CRP to monocytes; and no significant binding to lymphocyte populations could be detected. These studies confirm the ability of human CRP to bind to a majority of human monocytes in a calcium-dependent and specific manner, and provide further support for a biologically important interaction of this acute-phase protein with phagocytic cells.
Glomerular subendothelial and subepithelial immune complexes, containing the same antigen, are removed at different rates
Authors: MANNIK M , STAPLETON S A , BURNS M W , ALPERS C E , GAUTHIER V J
Published: July 2010

To examine the persistence of immune deposits in the subendothelial and subepithelial areas of the glomerular basement membrane in rats, immune deposits were formed by injection of radiolabelled, cationized human serum albumin (HSA) as antigen, followed by rabbit antibodies to HSA. The disappearance of the radiolabelled antigen from immune deposits in glomeruli was described by a curve consisting of two exponential components. By electron microscopy, subendothelial and subepithelial immune deposits were initially present in glomeruli. At later time-points, only subepithelial immune deposits were present. The fast component of disappearance, attributed to subendothelial deposits, had a half-life of 3.89±0.32 h. The slow component of disappearance from glomeruli, attributed to subepithelial deposits, had a half-life of 85.5±3.1 h. Since some of the injected, radiolabelled antigen was sequestered in other compartments of the body, the possibility was raised that antigen from these sites might be released and contribute to the persistence of deposits in glomeruli. This possibility, however, was excluded when transplantation of kidneys with immune deposits to untreated recipients revealed no difference in the amount of antigen persisting in nontransplanted and transplanted kidneys.
Decreased immunoreactive beta-endorphin in mononuclear leucocytes from patients with rheumatic diseases
Authors: WIEDERMANN C J , SACERDOTE P , MUR E , KINIGADNER U , WICKER T , PANERAI A E , BRAUNSTEINER H
Published: July 2010

The neuroendocrine potypeptide hormone beta-endorphin (β-END), which is released from various tissues including the anterior pituitary gland and cells of the immune system, has recently been implicated as having an immunoregulalory role. We used a radioimmunoassay to measure β-END levels in circulating mononuclear leucocytes from normal subjects and patients with various rheumatic diseases. Levels of β-END in leucocytes from patients were lower than in leucocytes from healthy subjects (P<0.001). Whereas levels of β-END in leucocytes from patients with the various rheumatic disorders were not significantly different, an inverse correlation was found between β-END levels in leucocytes and expression of rheumatoid factor (P<0.025) and erythrocyte sedimentation rate (P<0.025). This study demonstrates decreased content of β-END in cells of the immune system related to parameters of inflammatory activity in rheumatic diseases.
In situhybridization of IL-6 in rheumatoid arthritis
Authors: WOOD N C , SYMONS J A , DICKENS E , DUFF G W
Published: July 2010

IL-6, an important mediator of the acute phase response, has been implicated in the pathogenesis of rheumatoid arthritis (RA). Many cell types including macrophages, T cells, B cells, endothelial cells and fibroblasts can produce this cytokine and production is largely regulated at the level of gene transcription or mRN A stabilization. In this paper we have first measured the levels of IL-6 activity in synovial fluid (SF) and serum from patients with RA and then localized IL-6-producing cells in the synovium by in situ hybridization combined with immunophenotyping. Patients with RA had raised levels of IL-6 in both SF and serum compared with patients with osteoarthritis and age-matched healthy controls. In individual RA patients tested serially after admission to hospital, serum IL-6 was initially raised and, unexpectedly, increased with clinical improvement, In situ hybridization of IL-6 mRNA showed positive cells both in the lymphocyte-rich aggregates and adjacent to small blood vessels. With immunophenotyping it was found that cells containing IL-6 mRNA were often in contact with CD14+ tissue macrophages and double immunophenotyping revealed that immunoreactive IL-6 was often associated with synovial T cells.
Mucosal cell-mediated immunity to mycobacterial, enterobacterial and other microbial antigens in inflammatory bowel disease
Authors: IBBOTSON J P , LOWES J R , CHAHAL H , GASTON J S H , LIFE P , KUMARARATNE D S , SHARIF H , ALEXANDER-WILLIAMS J , ALLAN R N
Published: July 2010

Culture studies have suggested that Mycobacterium paratuherculosis may play a role in the aetiology of Crohn's disease. However, evidence of sensilizalion to mycobactcrial antigens amongst patients with Crohn's disease has not yet been adequately demonstrated. Previous studies of cell-mediated immunity (CMI) in Crohn's disease were restricted to responses of peripheral blood mononuclcar cells (PBMC) to mycobacterial antigens. In this study we have investigated the proliferativc responses of both PBMC and mesenteric lymph node mononuclcar cells (MLNMC) to a range of mycobacterial and non-mycobactcrial antigens. There was no evidence of specific sensitization in the responses of MLNMC and PBMC from patients with inflammatory bowel disease (IBD) to the mycobacterial antigens. However, anergy to M. paratuherculosis could not be excluded. IBD MLNMC responses to most antigens were generally greater than those of PBMC. which were often undetectable. When compared with controls, there was evidence of increased CM 1 to a range of non-mycobactcrial antigens, especially Yersinia enterocolitica. amongst both MLNMC and PBMC from patients with Crohn's disease and ulcerative colitis (UC). These results do not provide support to the proposed role of mycobacteria in the pathogcnesis of Crohn's disease, but indicate that further investigation may determine a role for bacterial-specific T cell-mediated responses in the palhogcncsis of IBD.
Subclass composition and J-chain expression of the‘compensatory’ gastrointestinal IgG cell population in selective IgA deficiency
Authors: NILSSEN D E , BRANDTZAEG P , FRØLAND S S , KAUSA O
Published: July 2010

The subclass disiribution of IgG-producing immunocytcs was examined by two-colour immunohistochemistry in gastrointestinal mucosa of 14 patients with selective serum IgA deficiency providing the following biopsy material: gastric (n=1); jejunal (n=12); colonic (n=1); and rectal (n=2). All except two patients suffered from various infections, and coeliac disease was observed in six of them. Control reference data were based on biopsies from immunologically intact subjects, including histologically normal jcjunal (n= 10) and large bowel (n=10) mucosa and stomach mucosa with slight chronic gastritis (n= 8). The total mucosal population of immunoglobulin-producing cells per 500 μm gut length unit was only slightly decreased in IgA deficiency because of an increased number of IgG (30%) and especially IgM (71%) immunocytes. The IgG I immunocytc proportion in the proximal gut (median 87%) was higher than that in the comparable controls (gastric 69%, jejunal 66%). A similar trend was seen in the distal gut (69%) compared with controls from the large bowel mucosa (55%). Conversely. lgG2 and lgG3 cell proportions were significantly decreased compared with the respective controls from the proximal gut. The same was true for IgG4. which also was significantly reduced in jejunal mucosa. Paired staining for cytoplasmic J chain and immunoglobulin isotype showed 71% positivity for jejunal IgG-producing cells in IgA deficiency, which was somewhat reduced compared with comparable controls (89%). J chain appeared to be preferentially expressed by IgGl cells (75%). but was also found in lgG2 (70%). lgG3 (32%) and IgG4 cells (33%). IgM-producing cells showed a J-chain positivity (99%) in IgA deficiency similar to normal (100%). Our results suggested that the block in mucosal B cell differentiation to IgA expression in the proximal gut is mainly located immediately upstream to the CHαl gene, giving excessive terminal maturation of J-chain-positivc IgGl immunocytes.
The role of interferon-gamma in lymphocytic thyroiditis: its functional and pathological effect on human thyrocytes in culture
Authors: KUNG A W C , MA L , LAU K S
Published: July 2010

lnlerferon-gamma (IFN-γ) has been recognized to possess diverse non-immunological effects on epithelial cells such as cellular growth and differentiation. We have previously demonstrated that IFN-y suppressed thyroid-stimulating hormone (TSH(-stimulated thyroglohulin (TG) synthesis in human thyrocytes through inhibition of TG gene transcription. To define the pathological mechanism involved in the action of IF'N-y, we studied the ultrastructural changes of human thyrocytes cultured in monolayer. Stimulation of the thyrocytes with TSH 10 mU/ml for 2 days resulted in marked increase in TG release into the medium. This was accompanied by elongation of microvilli. increase in follicles and acinar formation, increase in secretory granules and prominence of Golgi apparatus and rough-surfaced cndoplasmic rcticulum. Addition of IFN-y (300 L ml) resulted in marked degeneration with shrinkage of the cell membrane, vacuolation of cytoplasm, swollen mitochondria and presence of lysosomal granules. C'o-culluring the thyrocytes with the IFN-y and TSH resulted in suppression of the morphological responsiveness to TSH. There was also suppression of TSH-induced TG secretion. However, at 500 U'ml IFN-y did not cause lysis of the thyrocytes as estimated by the cellular DN A content. Furthermore, binuclcatcd cells were frequently encountered in those wells that were treated with IFN-y for cither 2 or 5 days. The findings suggest that IFN-γ resulted in de-differentiation and degeneration of the thyrocytcs. which subsequently regained the growth potential and showed attempts at regeneration. This may explain why most patients with lymphocytic thyroiditis recover from the acute injury and do not suffer from permanent hypothyroidism.
Lymphocyte subsets and β2-microglobulin expression in chronic hepatitis C/non-A, non-B. Effects of interferon-alpha treatment
Authors: WEJSTÅL R , NORKRANS G , WEILAND O , SCHVARCZ R , FUCHS D , WACHTER H , FRYDEN A , GLAUMANN H
Published: July 2010

Thirty-three patients with chronic hepatitis C/non-A. non-B were included in a randomized controlled study of interferon-alpha 2b (IFN-α2b) treatment, 3 × 106 U three times weekly for 36 weeks. Using an immunoperoxidase technique, frozen liver biopsy specimens were examined with MoAbs for the presence of T helper cells (CD4), T suppressor/cytotoxic cells (CD8), total T cells (CD2) and B cells (CD22) before and after treatment, β2-microglohulin (β2-MG) expression on hepatocytes was semiquantified using a scoring system on sections from paraffin-embedded biopsy specimens. Serum levels of β2-MG were analysed with a radioimmunoassay technique. Intralobular T helper and T suppressor/cytotoxic cells declined significantly in the treated patients but not in the controls. The portal CD4/CD8 ratio did not change. Before treatment, serum β2-MG levels and hepatocyte β2-MG expression were significantly higher in patients with chronic active hepatitis compared to patients with chronic persistent hepatitis. Serum β2-MG levels increased significantly in responders during IFN treatment, with a maximum after 12 weeks. However, in the liver, the hepatocyte β2-MG expression was significantly decreased after treatment. Thus. IFN-α treatment does not seem to induce an increased HLA class I antigen hepatocyte expression in chronic non-A, non-B hepatitis, which favours the hypothesis that its anti-viral effects are more important in modulating the disease activity.
Antibodies to hepatitis C virus in essential mixed cryoglobulinaemia
Authors: DAMMACCO F , SANSONNO D
Published: July 2010

Although essential mixed cryoglobulinaemia (EMC) is recognized to be frequently associated with chronic liver disease, aetiology and pathogenesis of liver damage remain unsolved questions. The purpose of this study was to assess the possible causative role of hepatitis C virus (HCV) in the liver impairment occurring in patients with EMC. Twenty-six consecutive EMC patients were evaluated. All patients underwent percutaneous liver biopsy. Anti-HCV antibodies were assayed by ELISA and supported by a recombinant immunoblotting assay (4-RIBA). The prevalence of anti-HCV antibodies in patients with and without chronic active liver disease (CALD) was compared. Anti-HCV antibodies were detected in 13 patients (50%) by ELISA and confirmed in 11 of them (42.3%) by 4-RIBA, the remaining two patients being indeterminate in the supportive assay. CALD correlated significantly with anti-HCV antibodies: indeed, 7/11 (63.6%) anti-HCV+ patients showed histological and clinical pictures of CALD, compared with 1/15 (6.6%) anti-HCV− patients (P<0.01). With the exception of the patient who was found to be HBsAg+, no liver tissue expressed hepatitis B virus-related antigens in the hepatocytes. Additional histological findings included discrete lymphoid aggregates in portal tracts, siderosis, fatty changes, hyperplasia of Kupffer cells. It can be concluded that chronic liver damage in EMC is frequently associated with anti-HCV antibodies. Although the cause of EMC remains unknown, this study has obvious implications for clarifying the etiology of associated CALD and further supports the therapeutic use of interferons in this disease.
High antibody levels to the mycobacterial fibronectin-binding antigen of 30–31 kD in tuberculosis and lepromatous leprosy
Authors: ESPITIA C , SCIUTTO E , BOTTASSO O , GONZÁLEZ-AMARO R , HERNÁNDEZ-PANDO R , MANCILLA R
Published: July 2010

Immunoblot assays showed that mycobacterial fibronectin-binding antigens are important targets of the humoral immune response in tuberculosis and leprosy. Using culture filtrate antigens of Mycobacterium tuberculosis, strong reactivity with the fibronectin-binding of 30–31 kD (Fn 30–31) was demonstrated in 55.9% of tuberculosis sera and in 56.5% of lepromatous leprosy sera. Sera from patients with tuberculoid leprosy and control sera gave very weak binding. Reactivity of tuberculosis and lepromatous leprosy sera with the fibronectin-binding antigen of 58–60 kD (Fn 58–60) was less conspicuous. The ability to react with fibronectin of the antigens of 58–60 and 30–31 kD was demonstrated by parallel labelling with a fibronectin-biotin conjugate. Fn 30–31 was purified to homogeneity by a two-step procedure and used for ELISA. Positive titres were found in 63% out of 65 tuberculosis sera and in 60.5% out of 43 lepromatous leprosy sera. Antibody titres in lepromatous leprosy sera were higher than in tuberculosis sera. Our observations indicate indirectly that M. leprae possess a highly immunogenic molecule homologous to M. tuberculosis Fn 30–31, which elicits a high antibody response in lepromatous leprosy but not in tuberculoid leprosy. In this investigation, direct evidence for the presence of this antigen in M. leprae was obtained by immunochemistry of lepromatous leprosy lesions with a monospecific antibody raised against M. tuberculosis Fn 30–31.
Detection of antibodies to human nerve antigens in sera from leprosy patients by ELISA
Authors: PARK J Y , CHO S N , YOUN J K , KIM D I , CELLONA R V , FAJARDO T T , WALSH G P , KIM J D
Published: July 2010

Anti-neural antibodies have been implicated to play a role in the pathogenesis of nerve damage in leprosy patients. To find the relationship between anti-neural antibodies and clinical findings, we attempted to detect antibodies against neurofilament-enriched proteins by ELISA in sera from leprosy patients. Of 289 sera from leprosy patients, 74(25.6%) had significant anti-neural antibodies: in contrast, 1 (5.0%) of 20 tuberculosis patients and 11 (7.1%) of 154 controls were seroreactive to nerve antigen. When clinical types were considered, a significant level of anti-neural IgG antibodies was detectable in 53 (30·1%) of 176 sera from lepromatous patients compared with 21(18.6%) of 113 sera from tuberculoid patients, indicating that lepromatous patients were more likely to be seropositive to nerve antigens in ELISA. Some of the ELISA-reactive sera showed antibody reactivity with 38-kD, 40-kD and 43-kD nerve antigens in Western blotting analysis. There was no apparent correlation between seroreactivity to nerve antigens and bacterial load in leprosy patients. Although there was no statistical significance, anti-neural antibodies were detectable more often among the patients on chemotherapy than the untreated and among the patients with erythema nodosum leprosum than without. The results, therefore, suggest that anti-neural antibodies arc elicited during the course of leprosy and may be associated with the extensiveness of nerve involvement in the patients.
An association between homozygous C3 deficiency and low levels of anti-pneumococcal capsular polysaccharide antibodies
Authors: HAZLEWOOD M A , KUMARARATNE D S , WEBSTER A D B , GOODALL M , BIRD P , DAHA M
Published: July 2010

Inherited deficiencies of complement components are associated with an increased risk of infection by encapsulated, high grade bacterial pathogens such as Streptococcus pneumoniae, Haemophilus influenzae type b and Neisseria meningitidis. Hence, the levels of antibodies to bacterial capsular polysaccharide antigens were measured using ELISA in 65 patients with inherited deficiencies covering the classical, alternative and terminal components of the complement cascade. Three of the four C3-deficient individuals studied were found to be almost totally deficient in specific anti-pneumococcal capsular polysaccharide (PCP) antibodies. These individuals had a history of recurrent pneumococcal sepsis. While single individuals with Clr, C2 and ClInh deficiency were found to have low anti-PCP antibody levels, no other group of complement deficiency had significantly reduced anti-PCP antibody levels compared with 100 controls. Antibody levels to the other two polysaccharides were not significantly lower in the patient groups. These findings suggest that C3 may be able to provide a stimulatory signal to promote the production of anti-PCP antibodies.
Evidence for the presence of activated CD4 T cells with naive phenotype in the peripheral blood of patients with rheumatoid arthritis
Authors: MAURER D , FELZMANN T , HOLTER W , PETERA P , SMOLEN J , KNAPP W
Published: July 2010

We have investigated whether T cell activation in rheumatoid arthritis (RA) preferentially engages distinct T cell subpopulations in the peripheral blood (PB) and in the synovial fluid. We found that CD25 expression was enhanced among PB CD4 T cells of RA patients as compared with CD4 cells of patients with reactive arthritis, degenerative joint disease or of healthy controls. Within the CD4 T lymphocyte subset we found that the CD45RO− (naive) cells selectively in RA displayed higher levels of CD25 protein and of interferon-gamma mRNA expression when compared with the respective subset of all other investigated groups. These results show that in the PB of RA, but not in the PB of the other arthropathies or healthy controls, CD45RO− CD4 T lymphocytes exist which display well-defined signs of activation.
Regulation of IL-2 production by mononuclear cells from rheumatoid arthritis synovial fluids
Authors: FLESCHER E , BOWLIN T L , TALAL N
Published: July 2010

Products of polyamine oxidation down-regulate IL-2 production by peripheral blood T cells. We show here that the production of IL-2 by rheumatoid arthritis synovial fluid mononuclear cells is inversely correlated with the concentrations of polyamines in these cells. In addition, the inhibition of polyamine biosynthesis or oxidation in cultures of these cells enhances their ability to produce IL-2. Our findings suggest that polyamine oxidation plays an important role in the suppression of T cell function characteristic of rheumatoid arthritis synovial fluids.
Effect of 12,13-phorbol dibutyrate and ionomycin on defective B cells in common variable immunodeficiency
Authors: FRANZ A , BRYANT A , WEBSTER A D B , FARRANT J
Published: July 2010

Secretion or IgM and IgG in vitro by B cells from patients with common variable immunodeficiency (CVI) has been used to classify the disease into three groups. On stimulation with anti-IgM and IL-2, group A patients' cells fail to secrete IgM or IgG, group B patients' cells secrete no IgG and significantly lower levels of IgM than normal cells, and group C patients' cells produce normal levels of both isotypes, Direct activation of protein kinase C using 12, 13-phorbol dibutyrate and ionomycin followed by IL-2 or IL-4 has been reported to induce immunoglobulin secretion by normal human B cells. We therefore attempted to induce B cells from group A and group B CVI patients to secrete IgM and IgG after direct activation of protein kinase C together with IL-2 or IL-4. The data show that the failure of secretion of immunoglobulin by B cells from CVI patients could not be reversed using this approach. This finding suggests that the activation channel involving protein kinase C in B cells from CVI patients is not involved in the defect in cell differentiation.
IL-2 infusion abrogates humoral immune responses in humans
Authors: GOTTLIEB D J , PRENTICE H G , HESLOP H E , BELLO C , BRENNER M K
Published: July 2010

Although IL-2 infusion enhances cell-mediated cytotoxicity in patients with neoplastic disease, administration is paradoxically associated with a modest fall in total serum IgG and an increased risk of infection. We now show that the adverse effects of IL-2 infusion on the humoral immune system are substantial. Although IL-2 induces the B cell growth and differentiating factors IL-4 and IL-6, infusion abrogates primary antibody responses entirely and reduces secondary antibody responses 50-fold following antigen challenge. There is no evidence of the generation of cells with suppressive activity on B cells but IL-2 increases the ratio of circulating virgin: memory cells. These results may help to explain the increased rate of bacterial infection in patients receiving IL-2. As IL-2 plays a central role in the generation of an immune response, the finding that it is also sufficiently immunosuppressive to inhibit primary- and secondary-type antibody responses suggests that exploration of the underlying mechanisms may provide insights into immune system homeostasis and may offer new approaches to therapeutic immunosuppression.
Up-regulation of tumour necrosis factor-alpha receptors on monocytes by desferrioxamine
Authors: PHILIPPE C , FOUQUERAY B , PEREZ J , BAUD L
Published: July 2010

The effect of endogenously generated reactive oxygen metabolites on the interaction of human blood monocytes with tumour necrosis factor-alpha (TNF-α) was investigated. Pre-exposure of unactivated human blood monocytes to dimethylthiourea, a scavenger of hydroxyl radical (OH˙) or to desferrioxamine (DFX), an iron chelator preventing the synthesis of OH˙, enhanced the specific binding of 125I-TNF-α to its receptors. Scavengers of superoxide anion or hydrogen peroxide were without effect. DFX-induced up-regulation of 125I-TNF-α binding depended on the concentration of the drug (1–5 mm) and on the duration of the treatment (1–18 h). It was not due to a reduction of receptor occupancy by endogenously generated TNF-α. Scatchard analysis of binding data revealed that DFX caused an approximately two-fold increase in the number of type II TNF-α receptors, with no change in their affinity. This up-regulation, that did not require synthesis of new proteins, was associated with a decrease in the internalization rate of TNF-α receptors, the half-life of which was doubled. Conversely, these findings suggest that OH˙ generation by monocytes may have a physiological role in reducing the activity of membrane-associated TNF-α receptors.
Status of immune-based therapies in HIV infection and AIDS
Authors: FAHEY J L , SCHOOLEY R
Published: July 2010

As our understanding of HIV disease pathogenesis progresses, approaches to immune-based therapy are evolving. Initial therapies aiming to alter immune function in patients with HIV infection have had mixed results. Clinical benefit in the trials so far has not been dramatic, although the studies are still at an early stage, and the correct protocols for the various agents or combinations of agents have yet to be established. As might be expected, where apparent benefit has occurred, it has been more obvious in those patients whose immune system was still intact.
Inhibitory activity of HIV envelope gp120 dominates over its antigenicity for human T cells
Authors: MANCA F , WALKER L , NEWELL A , CELADA F , HABESHAW J A , DALGLEISH A G
Published: July 2010

HIV-1 envelope glycoprotein (gp120), as a CD4-binding reactant, has been shown to inhibit in its native form human T cell responses to several antigens. Here we show that gp 120 in soluble form also inhibits activation of a specific human T cell line that responds to gp120-pulsed autologous antigen-presenting cells. In addition the inhibitory property of gp120 for antigen-driven T cell proliferation depends upon its ability to bind CD4 and is lost when CD4-binding capacity is abolished by denaturation, or blocked by complexing with soluble CD4 or with polyclonal antibodies. In contrast, antigenicity of denatured or complexed gp120 for specific human T cells is preserved. Similar effects are also observed with another CD4-binding reactant (i.e. anti-Leu 3a MoAb), which stimulates and/ or inhibits human T cells specific for mouse immunoglobulins depending on native or denatured conformation.
Impaired phagocytosis ofStaphylococcus aureusby granulocytes and monocytes of AIDS patients
Authors: POS O , STEVENHAGEN A , MEENHORST P L , KROON F P , VAN FURTH R
Published: July 2010

In the present study the microbicidal activities of granulocytes and monocytes from AIDS patients (CDC group IV) were assessed and compared with those of healthy controls. The phagocytosis and intracellular killing of Staphylococcus aureus by patient and control cells were measured using a method in which the rate of intracellular killing can be assessed independently of the rate of phagocytosis. Both granulocytes and monocytes of AIDS patients showed a decreased phagocytosis of S. aureus in comparison to phagocytes of healthy individuals. The rates of intracellular killing of S. aureus by granulocytes and monocytes did not differ significantly between these patients with late-stage HIV infection and controls.
Anti-mitochondrial antibody IgG subclass distribution and affinity in primary biliary cirrhosis
Authors: ZHANG L , WEETMAN A P , JAYNE D R W , TURNER I , YEAMAN S J , BASSENDINE M F , OLIVEIRA D B G
Published: July 2010

We have studied the total IgG subclass and anti-mitochondrial antibody (AMA) specific IgG subclass distribution in primary biliary cirrhosis (PBC) sera. In order to solve the problems caused by the differing affinities of subclass specific monoclonals and the competitive inhibition of antibodies in a whole serum assay, six sera were separated into subclass-specific fractions by affinity-depletion chromatography. AMA subclass distribution of 20 further sera from patients with PBC was assessed using conventional methods and the results were calibrated against one of the fractionated sera. Light chain distribution and AMA functional affinity were also assessed for the fractionated subclasses. Total amounts of IgG3 were significantly increased compared with normal controls. AMA were found in all IgG subclasses and not restricted predominantly to IgG3 as previously described. The functional affinity of IgG3 AMA is generally lower as compared with that of other subclasses. No light chain restriction was found.
Different mechanisms by which anti-DNA MoAbs bind to human endothelial cells and glomerular mesangial cells
Authors: CHAN T M , FRAMPTON G , STAINES N A , HOBBY P , PERRY G J , CAMERON J S
Published: July 2010

The mechanisms by which anti-DNA MoAbs derived from MRL-lpr/lpr mice, bind to human umbilical vein endothelial cells (HUVEC) and glomerular mesangial cells were studied using a cellular ELISA. DNAse-treatment of either the MoAb or HUVEC followed by reconstitution with DNA and/or histones was performed to determine whether DNA and histones mediated such binding. It was found that Mo Ab 410 bound to H U VEC and mesangial cells in the form of preformed DNA/anti-DNA immune complex, and such binding was facilitated by histones. In contrast, MoAb 152 bound directly to cell membrane-associated DNA, and adding DNA to MoAb 152 reduced its cellular binding. DNA binds endothelial cell surface and histones enhance the binding of both MoAb 410 and MoAb 152 to HUVEC by increasing cell membrane-associated DNA. Finally, the degree of MoAb binding to HUVEC is critically influenced by the relative concentrations of antibody, DNA, and histones.
Analysis of autoantibody production in SCID-systemic lupus erythematosus (SLE) chimeras
Authors: ASHANY D , HINES J , GHARAVI A , MOURADIAN J , ELKON K B
Published: July 2010

Mice with SCID disease have previously been successfully engrafted with human peripheral blood mononuclear cells (PBMC) obtained from normal individuals and from patients with various diseases. To determine whether SCID mice engrafted with SLE PBMC produced autoantibodies with specificities similar to those in the SLE donor, and to investigate which variables influence autoantibody production in the SCID recipients, we injected PBMC from 16 SLE patients into SCID mice and tested the recipients for autoantibodies to DNA and to five recombinant autoantigens. Ten out of 16 (68%) lupus and six out of nine (67%) normal grafts were successful as determined by the presence of human IgG ± 5 μg/ml of SCID serum post-transfer. Autoantibodies to La/SSB, Ro/SSA, and RNP were detected in five out of 10 SCID-SLE recipients by ELISA and immunoblotting up to 22 weeks post-engraft men I. The detection of autoantibodies in SCID-SLE mice was more closely related to autoantibody levels in donor sera than to total IgG concentrations in the SCID recipients. Autoantibody activity/mg IgG was similar in the donor and recipient sera. Histological evaluation of eight SCID-SLE mice killed 4–22 weeks post-transfer revealed population of the SCID thymus and spleen with mononuclear cells, but no evidence of lupus nephritis or dermatitis. These findings indicate that SCID mice can be engrafted with PBMC from patients with lupus and that specific autoantibodies are produced up to 5 months post-transfer. Failure to develop glomerulonephritis may be explained by low or absent anti-DNA antibodies or by changes in the cellular composition of the PBMC grafts.
Induction of autoantibodies to rat corneal protein 54
Authors: VERHAGEN C , BREEBAART A C , SUTTORP-SCHULTEN M S A , HOEKZEMA R , VAN HAREN M , KIJLSTRA A
Published: July 2010

Many studies have described the presence of circulating antibodies against corneal components in patients with corneal disease or uveitis, and in patients with skin or systemic disease with or without ocular involvement. The role of such antibodies in the underlying immunopathological process remains obscure. Here we describe the induction of autoantibodies against the rat cornea. Our attempts to induce corncal autoantibodies by various forms of keratitis and corneal trauma failed. However, circulating corneal autoantibodies could be detected by Western blotting after immunization of BN rats and Lewis rats with bovine corneal protein 54 (BCP 54). Rats immunized with rat corneal extracts (RaCE) or human serum albumin (HSA) as (auto) antigen did not develop corncal autoantibodies. During the study period (> 4 months), it was observed that the presence of circulating corncal autoantibodies did not elicit corneal inflammation. Severe keratitis did develop when BCP 54-immunized rats were challenged intracorneally with BCP 54, but the clinical signs were not significantly different from HSA-immunized rats after an intracomcal HSA challenge. Injection of corneal autoantibodies into the corneal stroma did not provoke kcratilis. To the best of our knowledge this is the first study demonstrating corneal autoantibodies in rats without actual manipulation of the eye. This model may provide further insights in the role and significance of corneal autoantibodies in disease.
Effects of haemorrhage on bacterial antigen specific pulmonary plasma cell function
Authors: ROBINSON A , ABRAHAM E
Published: July 2010

Nosocomial pneumonia is frequent after haemorrhage and trauma, and often contributes to multiple organ system failure, morbidity and mortality in this setting. Although the percentages and numbers of bacterial polysaccharide antigen-specific pulmonary B cell clonal precursors arc markedly decreased after haemorrhage, the effects of haemorrhage on pulmonary plasma cells actually producing antibody to these antigens are unknown. To investigate this question, the numbers of intraparcnchymal pulmonary plasma cells producing antibody against the bacterial polysaccharidc antigen levan (from Aerobacter levanicum) as well as bacterial antigen specific secretory IgA (sIgA) litres in the lungs were determined at various time points after 30% blood volume haemorrhage. Reduced numbers of bacterial antigen specific pulmonary plasma cells were found for more than 21 days following haemorrhage. An almost complete disappearance from the lungs of levan specific plasma cells occurred between 3 and 21 days after blood loss. Titres of bacterial antigen specific slgA in the lungs were decreased starting at 3 days post-haemorrhage and remained significantly depressed for more than 35 days after blood loss. These results demonstrate that haemorrhage produces profound and long-lasting suppression in bacterial antigen-specific pulmonary plasma cell function. Because these effects do not occur immediately post-haemorrhage, immunization techniques able to enhance bacterial antigen specific slgA litres at pulmonary surfaces may be able to increase resistance to nosocomial pneumonia if administered shortly after injury and blood loss.
Functional heterogeneity of human T cell clones from atopic and non-atopic donors
Authors: CHAMBERS C A , ZIMMERMAN B , HOZUMI N
Published: July 2010

Several distinct T helper (TH) subsets have been identified, based on the cytokines secreted. Recently, it has been demonstrated that these subsets can regulate the isotype of humoral responses. To investigate the possible differences in TH subsets between atopic donors which have elevated serum IgE and donors with normal serum IgE, we examined series of human TH cell clones. A total of 31 and 22 CD4+ T cell clones from the atopic and non-atopic donors, respectively, were characterized for the ability to help for IgE synthesis in vitro. T cell clones generated with allergen AgE from the atopic donor were autoreactive and all induced IgE synthesis. Tetanus loxoid-specific (TT) and phytohaemagglutinin clones were generated from both donors. There was significant heterogeneity between the T cells isolated with different stimuli from the same atopic donor. Also, there was a significant difference in the number of T cells generated from the atopic versus the non-atopic donor which helped for IgE, although there was no significant difference between the total number of T cells able to help for immunoglobulin synthesis of other isotypes. Most importantly, there was a higher frequency of clones able to support IgE synthesis between TT-specific T cell clones generated from the atopic versus the non-atopic donor. These results suggest that there are changes in subsets of TH cells specific for microbial antigens as well as allergens in atopics, which may have important implications for the aetiology of atopic disease.
Myelin antigen reactive T cells in cerebrovascular diseases
Authors: WANG W Z , OLSSON T , KOSTULAS V , HÖJEBERG B , EKRE H P , LINK H
Published: July 2010

T cell reactivities to the putative autoantigens myelin basic protein (MBP), MBP pcptides with amino acid residues 110–128 and 148–165, and myelin proteolipid protein (PLP) were examined in patients with acute ischaemic cerebrovascular disease (CVD) and, for comparison, in patients with inflammatory neurological diseases and other neurological diseases. A quantitative measure of these T cell reactivities was obtained by assessing numbers of T cells among blood and cerebrospinal fluid (CSF) mononuclear cells that secreted IFN-γ in response to antigen in vitro. Higher numbers of T cells reactive with each of these four antigens were detected in peripheral blood from patients with CVD compared with patients of the two control groups. Among blood cells from the CVD patients, their average number was 2·3–4·2/105 mononuclear cells. MBP reactive T cells were several-fold enriched in the CSF of CVD patients. The findings strongly suggest that brain damage in context with acute CVD leads to an in vivo expansion of myelin reactive T cells.
Reduction of graft-versus-host reactivity after small bowel transplantation: ex vivo treatment of intestinal allografts with an anti-T cell immunotoxin
Authors: CLARK C L INGHAM , SMITH G J , CRANE P W , PRICE B A , LEAR P A , FABRE J W , WOOD R F M
Published: July 2010

A specific T lymphocyte immunotoxin was used to pre-treat small bowel grafts in an attempt to prevent graft-versus-host (GVH) reactivity and GVH disease in a rat transplant model. The immunotoxin used was a conjugate of the anti-CD5 MoAb MRC OX-19 with ricin A chain. The grafts were perfused ex vivo with a standard solution of immunotoxin followed by incubation at 4°C for 1 h before transplantation. In a semi-allogeneic strain combination (parent to F1 hybrid offspring) graft treatment with immunotoxin led to a prolongation of recipient survival compared with groups receiving similar transplants without immunotoxin treatment. An additive effect on survival was observed when the host was treated with cyclosporin. The effect of immunotoxin was greater than that of mensenteric lymphadenectomy in increasing host survival. The effect of graft treatment with the immunoloxin on cellular migration from graft to host lymphoid tissues was assessed in fully allogeneic transplantation (PVG to DA). Host lymphoid tissues were subjected to immunohistochemical analysis using a MoAb specific for donor class I MHC antigens. Graft treatment with the immunotoxin led to a significant decrease in the number of graft cells found in host lymphoid tissues 7 days after transplantation. However, this effect was less marked than that achieved by graft mesenteric lymphadenectomy. With our current protocol graft treatment with a specific T cell immunotoxin can significantly reduce but not abolish GVH reactivity in rat small bowel transplantation.
Intracellular hydrogen peroxide production by peripheral phagocytes from diabetic patients. Dissociation between polymorphonuclear leucocytes and monocytes
Authors: NORITAKE M , ATSURA Y K , SHINOMIYA N , KANATANI M , UWABE Y , NAGATA N , TSURU S
Published: July 2010

Although the standard assays for reactive oxygen species have been based on the measurement of those released into the extracellular environment, the microbicidal capacity to the engulfed microorganisms is mainly dependent on those released into the intracellular environment, such as phagosomes. We studied intracellular oxidative activities of individual phagocytes by dichlorofluorescein (DCFH) oxidation assay to investigate the relationship between the reactive oxygen species released intracellularly and the impaired microbicidal capacity in diabetic patients. Time courses of intracellular production of hydrogen peroxide by polymorphonuclear leucocytes (PMNL) and monocyies were observed at the resting condition and after the stimulation with phorbol myristale acetate (PMA: 160 nm) by flow cytometry. Thirty-four patients with non-insulin-dependent diabetes mellitus (NIDDM) and 23 age-matched healthy volunteers were subjected to the studies. PMNL from patients with NIDDM showed a significantly decreased capacity to produce hydrogen peroxide after the stimulation (P < 0·05 at 15 min, P < 0·01 at 30 and 45 min). By contrast, inlracellular hydrogen peroxide production by monocytes at the resting condition and an early stimulatory phase (8 min after the stimulation) was significantly (P< 0·01) enhanced in patients with NIDDM compared with that in controls. Both the changes of inlracellular hydrogen peroxide production observed in PMNL and monocytes from patients with NIDDM were in association with an increased haemoglobin Alc level in crythrocytes. but did not relate to total cholesterol and triglyceride levels in the serum. The possible mechanisms of these dissociated changes in hydrogen peroxide producing capacity of phagocytes from patients with NIDDM are discussed.
Altered production of IgE and IgA induced by IL-4 in peripheral blood mononuclear cells from patients with IgA nephropathy
Authors: YANO N , ENDOH M , MIYAZAKI M , YAMAUCHI F , NOMOTO Y , SAKAI H
Published: July 2010

In order to elucidate the factors responsible for altered immunoglobulin production in patients with IgA nephropathy (IgAN). the in vitro effects oHL-4 and intcrferon-gamma (IFN-γ) on the synthesis of IgE and IgA by peripheral blood mononuclear cells (PBMC) were studied. Spontaneous IgE and IgA synthesis by PBMC was significantly increased in patients with IgA nephropathy compared with controls. The maximum amounts of IgA and IgE synthesis by PBMC after stimulation with IL-4 were almost the same both in patients with IgAN and in controls. The enhancement rate of IL-4-induccd IgE and IgA synthesis was significantly lower in IgAN than in the controls, suggesting in vivo preactivation of PBMC in IgAN patients. IFN-γ suppressed IgA and IgE synthesis by PBMC from IgAN patients as well as controls. However, the suppressive effect on IgE synthesis was less prominent in patients with IgAN. These results suggested that altered IL-4 action might be involved in the development of IgA ncphropathy.
Circulating immune complexes may play a regulatory and pathogenic role in experimental autoimmune uveoretinitis
Authors: KASP E , STANFORD M R , BROWN E , COOMBES A G A , DUMONDE D C
Published: July 2010

We compared the time course of changes in serum levels of circulating immune complexes (CICs) and of IgG antibody after sensilization of albino Lewis and pigmented Lister strain rats with uveitogenic (retinal S-antigen) and non-uveitogenic (ovalbumin) protein antigens of comparable molecular weight. Normal levels of CICs were far lower in Lewis rats in which experimental autoimmune uveoretinitis (EAU) lakes the form of a severe panuveitis, than in Lister rats, in which the disease is mild, focal, confined to the posterior segment, and of lower incidence. After sensitization with either S-antigen or ovalbumin, polyethylene-glycol-precipitable CIC (PEG-CIC) peaked and fell as IgG antibody levels rose in both rat strains. However, peak levels of PEG-CIC were lower and subsequent IgG antibody levels were higher in the Lewis strain than in the less susceptible Lister strain. In both strains of rat these linked PEG-CIC/IgG antibody responses occurred earlier after sensilization with uveitogenic (S-) antigen than with ovalbumin. whether or not individual S-antigen-sensitized Lister rats developed EAU. In contrast, complement-binding CIC rose substantially only in those rats of both strains displaying EAU in response to S-antigen and not in response to ovalbumin. We suggest that immune complex (idiotypic) regulation of IgG antibody responses may be more readily perturbed by a pathogenic autoantigen (S-antigen) than by a bland antigen (ovalbumin). We also suggest that differences between the balance of regulatory and pathogenic CIC responses to uveitogenic retinal antigen may underlie or reflect strain differences in susceptibility to and severity of EAU.
Expression of ICAM-1 (CD54) on normal and leukaemic B cells: implication for the mixed lymphocyte reaction
Authors: PADROS M R , NOLI M I , FAINBOIM L
Published: July 2010

Peripheral blood normal B lymphocytes were found to be poor stimulators in the mixed lymphocyte reaction (MLR), in contrast to normal activated B cells which were strong stimulators. This increased capacity to stimulate a strong MLR correlated with an increased expression of the ICAM-1 (CD54) molecule on the surface of these cells. Similarly, the capacity of leukaemic B cells to induce an allogenic stimulation in the MLR was limited to the ICAM-1 (CD54) positive leukaemic cells. The ability of normal activated or leukaemic B cells to induce an M LR was blocked by antibodies directed against ICAM-1.
Isolation and characterization of dendritic cells from adenoids of children with otitis media with effusion
Authors: VAN NIEUWKERK E B J , VAN DER BAAN S , RICHTERS C D , KAMPERDIJK E W A
Published: July 2010

Dendritic cells were enriched from adenoids of children with otitis media with effusion (OME) by density gradient centrifugation and culture techniques. An enrichment of 40–140-fold was obtained for dendritic cells. These cells were identified using morphology, enzyme cytochemistry, immunocytochemistry and functional criteria. Dendritic cells could be easily distinguished from macrophages. It appeared that the MoAb EBM 11 (CD68) discriminated between dendritic cells and macrophages; in dendritic cells this activity was localized in a spot, whereas in macrophages it was found throughout the whole cytoplasm. The fractions enriched with dendritic cells showed a strong stimulatory effect on allogeneic T cells. These responses were MHC class II dependent since they could be blocked by anti-HLA-DR/DQ MoAbs. The data clearly show that dendritic cells from adenoids of children with OME still have functional capacities.
Induction of endogenous retroviral gene product (SU) as an acute-phase protein by IL-6 in murine hepatocytes
Authors: ITOH Y , MARUYAMA N , KITAMURA M , SHIRASAWA T , SHIGEMOTO K , KOIKE T
Published: July 2010

The effect of Lps locus and IL-6 on the production of SU (previously termed gp70), a mouse endogenous retroviral gene product, was studied. Back-cross studies using the progeny between (NZB × C3H/HeJ)F1 and C3H/HeJ mice indicate that the basal level of SU is not associated with the Lps locus on chromosome 4. Lipopolysaccharide (LPS) mitogen response-negative mice did not show the enhancement of serum SU production after LPS injection. Spleen cells from LPS-mitogen response-positive but not from negative mice showed increase of IL-6 synthesis in the presence of LPS. Since IL-6 may be involved in the production of serum SU, we tested the effect of IL-6 in a primary hepatocyte culture system. SU production was clearly enhanced in the presence of recombinant IL-6, indicating that IL-6 induced by LPS can enhance the expression of retroviral genome.