bluuebunny/crossref_metadata_2025 huggingface dataset:
The significance of serum soluble IL-2 receptor as a marker for active visceral leishmaniasis in Sicilian patients
Authors: VITALE G , REINA G , MANSUETO S , MALTA R , GAMBINO G , MOCCIARO C , D'AGOSTINO R , DIELI M , CILLARI E
Published: August 2010

Sera from nine Sicilian patients with confirmed visceral leishmaniasis (Leishmania donovani infantum; VL), at the moment of the diagnosis, during the course of the disease and after clinical recovery, were analysed for the concentration of soluble IL-2 receptor (sIL-2R). The results show that sIL-2R is a marker of disease activity, since it is in high concentration at the beginning of infection and returns to the normal range following successful chemotherapy. At the same time of serum analysis for sIL-2R, peripheral blood mononuclear cells (PBMC) of VL patients were stimulated with phytohaemagglutinin (PHA) or antigen and supernatant tested for IL-2 and interferon-gamma (IFN-γ) production. Data demonstrate that there is an inverse relation between concentration of IL-2 and IFN-γ in the supernatants and sIL-2R secretion in the sera.
Activation of normal neutrophils by anti-neutrophil cytoplasm antibodies
Authors: KEOGAN M T , ESNAULT V L M , GREEN A J , LOCKWOOD C M , BROWN D L
Published: August 2010

Anti-neutrophil cytoplasm antibodies (ANCA) are markers of systemic vasculitis for which a pathogenetic role has been postulated. We have examined the effect of these autoantibodies on the function of normal human neutrophils in vitro. In the presence of ANCA positive sera luminolamplified chemiluminescence was significantly increased compared to the values seen in the presence of normal or anti-double stranded DNA positive sera (P<0·01). Five of six ANCA positive F(ab)2 preparations also produced significant neutrophil activation as demonstrated by the chemiluminescence response. This response was totally abrogated by the addition of neutrophil cytoplasm extract, containing the ANCA antigen. Addition of inhibitors to the chemiluminesccnce system demonstrated that the chemiluminescence response was inhibited by azide and salicylhydroxamic acid and reduced by histidine, suggesting that the chemiluminescence response was due to activation of myeloperoxidase, with generation of singlet oxygen. The chemotactic response to f-Met-Leu-Phe, a bacterial chemotactic peptide, was significantly augmented in the presence of ANCA. Chemotaxis to zymosan-activated serum and chemokinesis was not affected. Phagocytosis was also unaffected. We propose that neutrophil activation and modulation of neutrophil migration by ANCA may be of pathogenetic significance in systemic vasculitis.
T lymphocyte expression of complement receptor 2 (CR2/CD21): a role in adhesive cell-cell interactions and dysregulation in a patient with systemic lupus erythematosus (SLE)
Authors: LEVY E , AMBRUS J , KAHL L , MOLINA H , TUNG K , HOLERS V M
Published: August 2010

Complement receptor 2 (CR2, CD21), the receptor for both the C3d,g portion of human complement component C3 and the Epstein-Barr virus, has been recently described on peripheral T cells. By using dual stain flow cytometric analysis, we have also observed that a peripheral T lymphocyte subpopulation of normal healthy donors bears CR2 in a range varying from 1·1 to 23/2% (mean 12·6%) of total CD3+ cells. T lymphocytes from nine patients with inactive SLE expressed CR2 in a similar range. Three patients with active SLE were also studied. One of them, having neuropathy and glomerulonephritis, displayed an expansion of the CR2+ T cell subpopulation which reached as much as 89% of total CD3+ cells. To examine potential functional roles of T cell CR2, cells from a Jurkat-derived CR2 expressing T cell line were found to bind in vitro to human CR2−, complement-coated K562 cell targets in a CR2- and complement-dependent fashion. Based on these studies, we hypothesize that CR2 might act to increase adherence of T cells to nucleated target cells bearing C3d,g, a function which may be relevant to cytotoxicity or other T cell activities requiring cell-cell interaction.
In vitroinduction of IgG anti-DNA antibody from high density B cells of systemic lupus erythematosus patients by an HLA DR-restricted T cell clone
Authors: MURAKAMI M , KUMAGAI S , SUGITA M , IWAI K , IMURA H
Published: August 2010

An HLA-DR restricted T cell clone (26G11) which recognized a lymphoid cell-derived autoantigen associated with DR4 molecule was shown to induce not only autologous but also allogenic DR4+ B cells to produce large amounts of antibodies of the IgG and IgM classes. Using the helper activity of this clone, we investigated the mechanism of anti-DNA antibody production in DR-matched patients with systemic lupus erythematosus (SLE). When cultured with 26G11 cells, B cells from DR-matched normal control subjects produced large amounts of IgM anti-DNA antibody, but did not produce IgG anti-DNA antibody which is thought to have a pathological role in SLE. In contrast, B cells from DR-matched patients with active SLE spontaneously produced a fairly large amount of IgG anti-DNA antibody, and the production was augumented by the T cell clone. Little IgG anti-DNA antibody was produced by the B cells of patients with inactive SLE in either the presence or absence of T cell clone. We next fractionated B cells into low density B (LD-B) and high density B (HD-B) cells by centrifugation on discontinuous Percoll density gradients. IgG anti-DNA antibody was spontaneously produced by LD-B cells of active SLE patients but not by those either of inactive SLE patients or normal controls. On the other hand, although IgG anti-DNA antibody was not spontaneously produced by the HD-B cells of both active and inactive SLE patients, it could easily be induced by their culture with the T cell clone. Our results clearly show the existence of IgG anti-DNA antibody-producing B cells in the peripheral blood of SLE patients irrespective of their disease activity and suggest that autoreactive T cells may play a pathogenic role in SLE through the induction of autoantibody production.
Human polymorphonuclear leucocytes stimulated by tumour necrosis factor-alpha show increased adherence to extracellular matrix proteins which is mediated via the CD11b/18 complex
Authors: THOMPSON H L , MATSUSHIMA K
Published: August 2010

The present study demonstrates that tumour necrosis factor (TNF) and FMLP, but not IL-1 or IL-8, enhanced the adherence of polymorphonuclear neutrophil (PMN) to fibronectin, an extracellular matrix protein. The adherence induced by FMLP was very rapid, within 5 min while the induction of adherence by TNF was much slower, reaching maximum at 60 min. TNF also enhanced an adhesion of PMN to other extracellular matrix proteins, such as laminin, collagen IV and gelatin II, but not to human serum albumin. Anti-CD18 MoAb completely inhibited the binding of TNF-stimulated PMN to fibronectin and partially inhibited the binding to laminin. Further investigation showed thai adhesion of TNF-stimulated PMN to fibronectin and laminin was inhibited by anti-CD 11b MoAb and to a lesser extent by CD11a MoAb. In contrast to TNF-stimulated PMN the binding of unstimulated PMN to fibronectin and laminin was only inhibited by anti-CD11a MoAb. Anti-CD11c had no effect on PMN adherence. These results suggest that unstimulated PMN adhere to extracellular proteins through the CD11a/18, while TNF-stimulated PMN adhere through the CD11b/18. These results suggest that TNF secreted at the site of inflammation may enhance the interaction of PMN with the extravascular environment through the CD11b/18 complex.
Secretory IgA are elevated in both saliva and serum of patients with various types of primary glomerulonephritis
Authors: ROSTOKER G , TERZIDIS H , PETIT-PHAR M , MEILLET D , LANG P , DUBERT J M , LAGRUE G , WEIL B
Published: August 2010

Secretory immunoglobulin A (IgA) was determined by means of an enzyme-linked immunosorbent assay (using as capture antibody an MoAb specific for secretory component) in saliva and serum from 46 patients with IgA mesangial nephritis (IgAGN), 36 with an idiopathic nephrotic syndrome (INS), 30 with an idiopathic membranous nephropathy (MGN) and 40 healthy controls. Secretory IgA levels were elevated in both saliva and serum of patients with primary glomerulonephritis (P<0·05; Mann-Whitney test) regardless of the hislological type of the primary glomerulonephritis. Salivary IgA1 and IgA2 levels were increased in the saliva of patients with IgAGN, INS and MGN (P<0·05; Mann-Whitney test). The monomeric/total IgA ratio, and interferon-gamma and soluble IL-2 receptor levels, in saliva did not differ between the patients and controls (P>0·05: Mann-Whitney test). We conclude that the mucosal immune system is activated in forms of glomerulonephritis other than IgAGN.
Isolation and characterization of the tubular basement membrane antigen associated with human tubulo-interstitial nephritis
Authors: YOSHIOKA K , HINO S , TAKEMURA T , MIYASATO H , HONDA E , MAKI S
Published: August 2010

The target antigen, a 54-kD glycoprotein (gp54). reactive with sera from patients with anti-tubular basement membrane (anti-TBM) nephritis, was isolated from collagenase-digested (CD) bovine TBM. The purified gp54 was shown to be non-collagenous by amino acid analysis, and to be a unique basement membrane component by amino-terminal sequencing. The nephritogenicity of gp54 was demonstrated by immunizing strain XIII guineapigs with purified gp54, and producing anti-gp54 antibody and tubulo-interstitial nephritis. Anti-gp54 antibody, affinity-purified from sera of patients with anti-TBM nephritis, bound by immunoblotting to 54-kD and, to a lesser extent, 48-kD components of partially purified human CD-TBM. Indirect immunofluorescence showed that gp54 was present in the basement membrane of proximal tubules of the kidneys of normal human, cow, rabbit, guineapig and Brown-Norway rat but not in Lewis rat. Immunoelectron microscopy revealed localization of gp54 along the interstitial side of the TBM and its association with interstitial collagen fibres. These results indicate that gp54 is the nephritogenic antigen involved in tubulo-interstitial nephritis, and is unique in chemical characteristics and localization in the kidney.
Urinary excretion of nitrite and nitrate in experimental glomerulonephritis reflects systemic immune activation and not glomerular synthesis
Authors: SEVER R , COOK T , CATTELL V
Published: August 2010

In immune-induced glomerulonephritis (gn), glomeruli (gl) synthesize nitric oxide (NO), and urinary nitrite (NO2−) excretion is increased. In mammals on a low nitrate (NO3−) diet, urinary NO3− is a measure of endogenous NO3− synthesis. Excretion is increased after administration of macrophage activators, reflecting induction of NO production. To determine whether increased urinary NO2− gn is due to glomerular synthesis we studied urinary NO2−/NO3− in accelerated nephrotoxic nephritis induced by preimmunization with rabbit immunoglobulin G (IgG), followed by rabbit anti-rat nephrotoxic globulin, and in control rats similarly preimmunized with rabbit IgG, but followed by normal rabbit serum. Both urinary NO2− and NO3− were increased by i.p. preimmunization with rabbit IgG (peak 463 ± 171 nmol NO2−/60·3 ± 9·4 nmol NO3/24 h, P<0·001 for both NO2− and NO3− compared with preimmunization levels). Repeat immunization with i.v. rabbit anti-rat nephrotoxic globulin (nephritic rats) or normal rabbit globulin (control rats) again increased urinary NO2− and NO3−. There was no statistically significant difference in urinary NO2 and NO3− levels between nephritic rats where globulin had nephrotoxic activity and the control rats injected with normal rabbit globulin, despite increased NO2−synthesis in ex vivo nephritic glomeruli after nephrotoxic globulin (7·9 ± 1·9 nmol/2000 gl/48 h; controls 3·2 ± 1·0 nmol/2000 gl/48 h). Thus neither urinary NO2− nor NO3− levels reflect local activation of the NO pathway in glomeruli. As reported for other stimulants, we show here that systemic stimulation with foreign antigen increased NO synthesis.
Changes in circuIating B cells and immunoglobulin cIasses and subcIasses in a healthy aged popuIation
Authors: PAGANELLI R , QUINTI I , FAGIOLO U , COSSARIZZA A , ORTOLANI C , GUERRA E , SANSONI P , PUCILLO L P , SCALA E , COZZI E , BERTOLLO L , MONTI D , FRANCESCHI C
Published: August 2010

The study of 87 adults of different ages, including 15 centenarians, selected for their healthy status, showed that profound changes of humoral immunity occur throughout life. In particuIar, a statistically significant age-reIated increase of the serum level of immunoglobulin cIasses (IgG and IgA but not IgM) and IgG subcIasses (IgG1, 2 and 3, but not IgG4) was detected. A parallel age-related decrease of circuIating B cells was also observed. The hypothesis of a complex derangement of B cell function and/or compartmentalization with age is put forward, together with the proposal that healthy centenarians (as representative of successful ageing) may be helpful in identifying the physiological age-reIated modifications of the immune system.
Association of T cell and macrophage dysfunction with surface gp120-immunoglobulin-complement complexes in HIV-infected patients
Authors: DANIEL V , SÜSAL C , WEIMER R , ZIMMERMANN R , HUTH-KÜHNE A , OPELZ G
Published: August 2010

The mechanism of CD4+ cell depletion and functional T helper cell inhibition in HIV-infected individuals is poorly understood. The present study demonstrates that immune complex-covered CD4+ cells arc associated with T cell inhibition and macrophage stimulation. We studied 30 patients with ARC/AIDS and 35 asymptomatic HIV haemophilia patients. Overall, 20 ± 3% of peripheral CD4+ lymphocytes were covered with gp 120 (range 0–94%). Gp120+ cells also exhibited surfacebound IgG (P=0.0001). IgM (P= 0.0001), and complement (P= 0.0001). Decreased in vitro lymphocyte proliferation was associated with the immune complex load of CD4+ cells. The higher the percentage of CD4+ gp120+ cells in the blood, the lower the T cell response in vitro (P=0.00l). Moreover, an association was found between immune complex-positive cells and plasma neopterin (P=0.01). Patients with increased plasma neopterin levels had decreased in vitro responses to pokeweed mitogen (PWM) (P=0.006), phytohaemagglutinin (PHA) (P=0.004), concanavalin A (Con A) (P=0.09), and anti-CD3 MoAb (P=0.03), and decreased CD4+ cell counts in the blood (P=0.006). Since maximally 1% of CD4+ lymphocytes are infected with HIV, T cell dysfunction and T cell depletion in HIV-infected patients may also be caused by the release of free gp120 that binds to uninfected CD4+ cells. Our data suggest that the functional inhibition and subsequent elimination of uninfected CD4+ lymphocytes with surface gp120 immunoglobulin complement complexes may be a pathomechanism in the manifestation of AIDS.
Tumour necrosis factor productionin vivoandin vitroin response toParacoccidioides brasiliensisand the cell wall fractions thereof
Authors: FIGUEIREDO F , ALVES L M C , SILVA C L
Published: August 2010

Tumour necrosis factor (TNF) was detected in scrum from mice challenged with Paracoccidioides brasiliensis. The scrum TNF levels of mice challenged with an avirulent strain were significantly higher than those of mice challenged with a virulent strain, and the same was observed for the TNF levels of mice challenged with a cell wall fraction (F1) from the two fungal strains. Fraction F1 consisted of chitin and β-glucan; but although the chitin contents were similar for the two strains, the avirulcnt strain allowed a greater content of β-glucan. The β-glucan, purified from both strains, increased serum TNF levels in an identical dose-dependent manner, whereas purified chitin did not induce scrum TNF levels. P. brasiliensis, the F1 fractions and β-glucan induced macrophages to secrete TNF in vitro. The differences in TNF levels, induced by the different fungal strains, were correlated with the β-glucan concentrations in the cell walls of both the avirulent and virulent strains of P. brasiliensis. These findings support a role for TNF in the pathogenicity of P. brasiliensis.
Immunosuppressive property of bromocriptine on human B lymphocyte functionin vitro
Authors: MORKAWA K , OSEKO F , MORIKAWA S
Published: August 2010

Bromocriptine (BRC), a dopamine ergot alkaloid, inhibits the release of pituitary prolactin (PRL). Hypoprolactinaemia induced in rat by treatment with BRC produces a similar immunosuppressive effect as observed in hypophysectomized rats. The effect of immunosuppression by the administration of BRC has been interpreted as the result of hypoprolactinaemia produced by BRC. However, the direct effect of BRC on lymphocyte function has never been evaluated. The purpose of this study was to investigate the in vitro effect of BRC on human B cell functions. Highly purified B cells from tonsil samples were isolated by Percoll density gradient from non-rosctted cells, and were used as target cells. BRC significantly suppressed the proliferative response of resting and activated B cells in vitro. It suppressed immunoglobulin generation of activated B cells. The inhibition of BRC was manifested in the early stage of the proliferation and differentiation of B cells. The conditioned medium from the polyclonal B cell mitogcn-stimulated B cell cultures did not contain PRL as determined by immunoradiometric assay. Treatment with low-dose cyclosporin A or FK506 in conjunction with BRC has proved more effective than either drug alone in suppression of B cell proliferation. Thus, the combined therapy of BRC and immunosuppressants may be effective with decreased toxicity for clinical use.
Immunohistochemical detection of co-localizing cytokine and antibody producing cells in the extrafollicular area of human palatine tonsils
Authors: HOEFAKKER S , van't ERVE E H M , DEEN C , van den EERTWEGH A J M , BOERSMA W J A , NOTTEN W R F , CLAASSEN E
Published: August 2010

In vitro experiments have documented the role of cytokines in the regulation of the human humoral immune response. Which cytokines arc operative in vivo and in which lymphoid compartment interactions between cytokine-producing T cells and antibody-forming B cells occur is still unclear. For that reason we studied human tonsils using immunohistochemical techniques. In tissue sections from tonsils in a resting stage after recurrent tonsillitis we observed cells producing IL-1 α and tumour necrosis factor-alpha (TNF-α) which were exclusively localized in the mantle zone of the follicle and in the extrafollicular area. Furthermore, a high frequency of interferon-gamma (IFN-γ)-producing cells was detected in the extrafollicular area, but not inside the follicles. Occasional IL-2- and IL-4-producing cells were found in the extrafollicular area. Immunohistochemical detection of antibody isotypes revealed that B cells, IgM-membrane-positive, were localized inside the follicles and mantle zones, whereas IgD-membrane-positive cells were mainly found in the mantle zones of secondary follicles. In contrast, plasma cells producing IgGl-4 and IgAl-2 were found in the extrafollicular area. No IgD and IgE antibody-forming cells were detected in tonsils, whereas IgM antibody-forming cells were detected in the xtrafollicular area. The co-localization of cytokine-producing cells and antibody-forming cells in human tonsil suggests that T-B cell interactions, required for B cell differentiation and isotype switching, take place in the extrafollicular area.
Antimitochondrial autoantibodies in anti-glomerular basement membrane disease
Authors: MARRIOTT J B , OLIVEIRA D B G
Published: August 2010

Anti-glomerular basement membrane (GBM) disease is characterized by the production of an autoantibody with very restricted specificity, with no evidence of polyclonal B cell activation. It was therefore surprising to find that in a solid-phase ELISA a proportion of anti-GBM sera showed significant binding to pyruvate dehydrogenase (PDH), a reactivity usually associated with the antimitochondrial autoantibodies (AMA) found in primary biliary cirrhosis (PBC). The specificity of this reactivity was confirmed by inhibition and competition experiments. The AMA found in anti-GBM sera were of much lower affinity than those found in PBC sera, and recognized a more restricted set of species (mainly the 55-kD and occasionally the 74-kD component of PDH). However, it was possible to block the binding in a Western blot of an anti-GBM serum to both the 55-kD and 74-kD species with F(ab')2 fragments prepared from a PBC serum. Although AMA have been found in diseases other than PBC, such diseases have usually been characterized by polyclonal B cell activation. The stimulus to the production of AMA in anti-GBM disease, and their significance in pathogenesis (if any), are unknown.
Surface expression of adenosine deaminase in mitogen-stimulated lymphocytes
Authors: MARTIN M , CENTELLES J J , HUGUET J , ECHEVARNE F , COLOMER D , VIVES-CORRONS J L , FRANCO R
Published: August 2010

Adenosine deaminase (ADA) expression on the surface of mitogen-stimulated lymphocytes was studied by flow cytometry. The gate for lymphocytes was located by cell size (forward scatter), cytoplasmic complexity (side scatter) and by expression of the markers CD2, CD4. CD8 and CD19. After mitogenic proliferation two populations appeared, one corresponding to non-stimulated cells, and the other consisting of larger cells which showed relatively high expression of adenosine deaminase on their surface. The increase was similar to that observed for CD71 expression, and paralleled the increase in 3H-thymidine incorporation. There was a correlation between ADA and CD71 expression (r= 0.92 for phytohacmagglutinin (PHA) and 0.97 for pokeweed mitogen (PWM)). These results suggest a role for ecto-adenosine deaminase in lymphocyte proliferation and/or triggering.
Recombinant gp120 induces IL-10 in resting peripheral blood mononuclear cells; correlation with the induction of other cytokines
Authors: AMEGLIO F , CAPOBIANCHI M R , CASTILLETTI C , CORDIALI FEI P , FAIS S , TRENTO E , DIANZANI F
Published: August 2010

Immutiological abnotmalities present in HIV-1-infected individuals often reflect an imbalance of cytokine production. The HIV-I gpl20 has the ability to induce a number of cytokines, and to enhance immunoglobulin release by normal peripheral blood mononuclear cells (PBMC) in vitro, in the absence of IL-2 production and of lymphoproliferation. This study provides evidence that gpl20 is a potent IL-10 inducer in normal PBMC cultures. The pattern of other cytokines induced by gp 120 includes interferon-alpha (IFN-a) and IFN-γ, tumour necrosis factor-alpha (TNF-α), IL-6, IL-Iα2 and IL-β, and not IL-2 and IL-4. These findings further define the pattern of cytokine release induced by gpl20 on human resting PBMC. Furthermore, the present findings roughly parallel those observed both in the sera of patients and in the mononuclear cells from HIV+ individuals early after infection, suggesting that gpl20 could be a good candidate as one of the agents responsible for cytokine dysregulation observed in HIV-1-infected individuals.
Increased CD45RO expression on T lymphocytes in mediastinal lymph node and pulmonary lesions of patients with pulmonary sarcoidosis
Authors: FAZEL S B , HOWIE S E M , KRAJEWSKI A S , LAMB D
Published: August 2010

Sarcoidosis is characterized by a cell-mediated response mediated by the activation of CD4′ T lymphocytes in an environment lacking adequate numbers of regulatory CD8 T lymphocytes. Immunohistological studies on frozen tissues have shown that sarcoid lesions have activated CD4 heiper/inducer T lymphocytes at the centre of granulomata, whereas lymphocytes at the periphery are mainly CD8 suppressor/cytotoxic cells. In this study we investigated the immunohistological distribution of CD45 isoforms of T cells in 29 paraffin-embedded sarcoid lesions in mediastinal and open lung biopsies. Ten of these were assessed quantitatively, with single-staining of serial sections demonstrating a predominance of CD45RO memory T lymphocytes in granulomata and intergranulomalous areas. Ratios of CD45RO;CD45RA T lymphocytes (or the ratio oi memory to naive T cells) were 420:1 for granulomata and 17-9:l for intergranulomatous areas of sarcoid lesions counted. This finding is compatible with the hypothesis that nearly all the lymphocytes present in sarcoid lesions have been previously activated, and selectively home to sarcoid lesions.
Immunosuppressive activity of bromocriptine on human T lymphocyte functionin vitro
Authors: MORIKAWA K , OSEKO F , MORIKAWA S
Published: August 2010

Bromocriptine (BRC), a dopamine type 2 agonist, prevents secretion of pituitary prolactin (PRL). BRC has been shown to impair lymphocyte responsiveness toward antigenic stimulation by decreasing serum PRL levels, Hypophyophysectomized induced by BRC produces a similar immunosuppressive effect, as observed in hypophysectomized rats, which is restored by the administration of PRL, Therefore, the immunosuppression induced by BRC has been interpreted as the result of hypoprolactinaemia. However, the direct mechanism of BRC in immune response has never been evoked. We recently reported that BRC has an immunosuppressive activity on human B lymphocyte function in vitro. In the present study we demonstrate that BRC suppresses T cell proliferation by means of blocking IL-2 production by T cells as well as mixed lymphocyte reaction (MLR) in a dosedependent manner. We could not detect the immunoreactive PRL activity in the conditioned medium from polyclonai T cell mitogen-stimulated T cell cultures. Then, the immunosuppressive activity of BRC on human T cell function appeared to be independent of its hypoprolactinaemic effect. Treatment with low-dose cyclosporin A(CsA) or FK506 in combination with BRC has proved more effective than either drug alone in suppression of T cell proliferation and CD25 antigen expression. Thus, the therapeutic applicalion of BRC in combination with immunosuppressants may enhance the immunosuppressive effect, while at the same time decreasing the toxicity.
IL-6 induces hepatic inflammation and collagen synthesisin vivo
Authors: CHOI I , KANG H-S , YANG Y , PYUN K-H
Published: August 2010

IL-6 regulates the synthesis of a broad spectrum of acute phase proteins in the liver. Also, it is involved in the pathogenesis of many fibrogenic diseases. To study ihe inflammatory effects of IL-6 on the in vetro human rIL-6, produced in Escherichia coli. was injected intraperitoneally into rats (25 μ/100 g body weight). The major fractopm of injected 11-6 was accumulated in the liver within 40 min, and the number of platelets was increased during 72 hafter injection. After 5 weeks of injection. the levels of serum glutamine pyruvic transaminase (GPT) and glutamic oxaloacctic transaminase (GOT) were not changed, but they were significantly elevated at 13 weeks of treatment. Meanwhile, scrum albumin levels were slightly decreased compared with those of controls. The same phenomena were observed in carbon tetrachloride-treated rats. Collagen synthesis was increased in the liver tissues and in the culture supernatants of hepatic lipocyles isolated from the rats treated with lL-6for 13 weeks. Histological analysis correlated well with biochemical analysis. At 5 weeks of treatment, only mild pathological changes were observed, but severe hepatocyte necrosis and the accumulation of fibres in necrotic area were developed in the liver of IL-6-treated rats after 13 weeks of treatment, confirming that hepatic inflammation and fibrosis were developed. IL-6 activities in the sera and in the culture supernatants of lipocytes from IL-6-treated rats were elevated compared with those in controls. These biochemical and pathological data indicate that IL-6 can induce hepatic inflammation, and it has important roles in the pathogenesis of fibrosis and diseases of the liver in vivo. In addition, these results will provide useful information for ihc clinical trials of IL-6.
Constitutive c-fos expression in osteoblastic MC3T3-E1 cells stimulates osteoclast maturation and osteoclastic bone resorption
Authors: KUROKI Y , SHIOZAWA S , SUGIMOTO T , KANATANI M , KAJI H , MIYAUCHI A , CHIHARA K
Published: August 2010

The effect of culture supernatants of c-fos-transfectcd MC3T3-El osteob[aslic cells on osteoclastic bone resorption was studied. Human c-fos cDNA was integrated in the expression vector pH8. and the cells were transfected using the calcium phosphate precipitation technique. Osteoclastic bone resorplion was quantified by the pit formation assay, and the osteoclast maturation from precursor was assessed by the generation of tartrate-resistant acid phosphatase (TRAP)-positive multinucleated cells (MNC). The culture supernatants of MC3T3-E1 transfectants constitulively expressing c-fos gene enhanced osteoclast-like MNC formation from haematopoietic blast cells compared with those of control transfectants (P<0·01). The culture supernatants also promoted osteoclastic bone resorption: the pit number. 118·7±38·5, was significantly higher than 19·0±10·1 of the control (P<0·05). The absorption area. 12394±3145 mm2, was significantly larger than 1646+314 mm2 of the control (p<0-05). The culture supernatants also promoted bone resorption by purified chick osteoclasts (P<0·05). The results show that constitutive expression of c-fos gene in osteoblastic MC3T3-EI cells stimulates osteoclast rnaturation and osteoclastic bone resorption by releasing humoral mediator(s).
Increased numbers of CD5+ B cells and T cell receptor (TCR) γδ+ T cells are associated with younger age of onset in rheumatoid arthritis (RA)
Authors: Hassan J , Yanni G , Hegarty V , Feighery C , Bresnihan B , Whelan A
Published: August 2010

Patients presenting with RA before the age of 45 years (younger onset) are known to have more aggressive disease compared with patients presenting after the age of 65 years (older onset). Coordinated expansion of circulating CD5+ B cell and TCR γδ+ T cell levels has been reported in patients with RA. This study assesses the peripheral blood levels of these two cell types in RA patients with younger and older onset of disease. CD5+ B cell levels were significantly elevated in the younger onset RA group (26±6 γδ+4±5%) compared with the older onset RA group (14±2 γδ+1±2%; P<0±01). TCR γδ+ T cell levels were also significantly raised in the young patients (4±0 γδ+0±9%) compared with elderly patients (1±6 γδ+0±2%; P<0±01). T cell levels (CD3+) were similar in both groups (young 66±4 γδ+3±3%; old 74±3 γδ+3±4% (mean γδ+s.e.m.); NS). Total B cell levels (CD19+) were also similar in these groups (7±7 γδ+0±7%versus 8±9 γδ+1±8%; NS). A significant positive correlation was observed between the CD5+ B and TCR γδ+ T cell types in the patients (r= 0±72, P<0.05). Compared with age-matched normal controls, the younger onset patients had similar CD5+ B cell and TCR γδ+ T cell levels to the elderly controls (CD5+ B cells 30±2 γδ+3±0%; TCR γδ+ T cells 3±0 γδ+0±8%). Conversely, older onset RA patients had CD5+ B cell levels similar to the young controls (12±3 γδ+1±9%). Spontaneous in vitro synthesis of immunoglobulins (IgM, IgA and IgG) and rheumatoid factors (IgM and IgA isotypes) were not significantly different in both patient groups. The coordinate expansion of circulating CD5+ B cells and γδ+ T cells seen in patients with RA presenting before 45 years of age and not after 65 years of age may suggest a potential role for these cells in more aggressive disease states.
Therapeutic effects of estradiol benzoate on development of collagen-induced arthritis (CIA) in the Lewis rat are mediated via suppression of the humoral response against denatured collagen type II (CII)
Authors: Waksman Y , Hod I , Friedman A
Published: August 2010

The effects of estradiol benzoate (EB) on the development of anti-CII antibodies and their pathogenic potential were studied during the progress of established CIA in the rat. CIA was induced in mature female Lewis rats by two subcutaneous inoculations containing bovine native CII (BCIIn), emulsified in Freund’s incomplete adjuvant. Clinical arthritis fully developed by day 18 and then EB (1 mg/kg body wt per day, diluted in corn oil (CO)) was administered intramuscularly every second day thereafter. Antibodies binding four different CIIs (bovine or rat, either native or heat-denatured) were detected in sera and joint tissue extracts by means of solid-phase ELISA. Pharmacological doses of EB (>0±2 mg/kg body wt per day) caused significant remission of established CIA 5-7 days after treatment, and selectively suppressed the production of antibodies specific for denatured CII. To evaluate the arthritogenic potential of circulating anti-CIId IgG, transfer experiments were performed. IgG anti-CIIn, purified from EB-treated CIA rats, was not arthritogenic, whereas IgG anti-denatured (CIId), purified from CO-treated CIA rats, caused severe passive arthritis. Furthermore, pretreatment with rat CIId protected against subsequent induction of CIA, and this protection was associated with suppressed antibody production against CIId. Collectively, our results indicate that antibodies specific for CIId are involved in the pathogenesis of CIA, and that oestrogen-related remission of clinical arthritis may be caused by a selective suppression of antibodies produced against degraded denatured CII.
Natural killer (NK) cell deficiency associated with an epitope-deficient Fc receptor type IIIA (CD16-II)
Authors: Jawahar S , Moody C , Chan M , Finberg R , Geha R , Chatila T
Published: August 2010

Susceptibility to herpes virus infections has been described in experimental animals depleted of NK cells and in patients with defective NK cell function. We have identified a child with recurrent infections, especially with herpes simplex virus, who had a decreased number of CD56+CD3− NK cells in circulation. Her NK cells expressed an altered form of the Fc receptor for IgG type IIIA (FcγRIIIA or CD16-II) which was not reactive with the anti-CD16-II MoAb B73.1. Sequence analysis revealed the patient to be homozygous for a T to A substitution at position 230 of CD16-II cDNA, predicting a Leu66 to His66 change in the first immunoglobulin domain of CD16-II at the B73.1 recognition site. Spontaneous NK cell activity of the patient’s peripheral blood mononuclear cells (PBMC) was markedly decreased, while antibody-dependent cellular cytotoxicity (ADCC) was unaffected. These results suggest that this child suffers from a defect affecting the development and function of NK cells, resulting in NK cytopenia and clinically significant immunodeficiency. The role of the CD16-II mutant in the pathogenesis of the patient’s NK cell deficiency is discussed.
Transforming growth factor-beta 1 (TGF-β1)-mediated inhibition of glial cell proliferation and down-regulation of intercellular adhesion molecule-1 (ICAM-1) are interrupted by interferon-gamma (IFN-γ)
Authors: Xiao B-G , Zhang G-X , Ma C-G , Link H
Published: August 2010

We utilized a model of myelin basic protein (MBP) activation in vivo and MBP-stimulated cultures in vitro to study the influence of TGF-β1 on glial cell proliferation and ICAM-1/leucocyte function-associated antigen-1 (LFA-1) expression, and to observe the antagonistic effects of TGF-β1 and IFN-γ. TGF-β1 inhibited MBP-stimulated and MBP-activated glial cell proliferation, especially in MBP-stimulated separated microglia and astrocytes, and down-regulated the expression of ICAM-1 on MBP-stimulated glial cells and separated microglia. ICAM-1 expression on MBP-activated glial cells was intensely suppressed, whereas its expression on MBP-stimulated astrocytes was not influenced. TGF-β1 had no effect on LFA-1 expression. In contrast, IFN-γ up-regulated ICAM-1 expression, but inhibited proliferative response on MBP-stimulated glial cells when cultured without TGF-β1. Examination of TGF-β1 and IFN-γ interactions revealed that TGF-β1-mediated inhibition of proliferation and down-regulation of ICAM-1 on glial cells were prevented by IFN-γ. The suppressive effect was re-established with high doses of TGF-β1 in cultures, indicating that biological effects of TGF-β1 vary depending on nitric oxide (NO) production, its concentration in the microenvironment and regulation of the cytokine network.
Prednisone increases apoptosis inin vitroactivated human peripheral blood T lymphocytes
Authors: Lanza L , Scudeletti M , Puppo F , Bosco O , Peirano L , Filaci G , Fecarotta E , Vidali G , Indiveri F
Published: August 2010

Glucocorticoid hormones (GCH) regulate, through the apoptotic process, the negative selection of immature T cells in the thymus. Because apoptosis seems to occur also in the maintenance of peripheral tolerance, we have investigated whether GCH may induce apoptosis in human mature lymphocytes. Peripheral blood lymphocytes (PBL) or peripheral CD4+ and CD8+ T cell subsets were cultured in the presence of phytohaemaglutinin (PHA) or PHA and prednisone (PDN) at 10−3-10−12M concentrations for 72, 96 and 120h. Cell cycle and membrane antigen expression were evaluated by flow cytometry and DNA degradation was detected by agarose gel electrophoresis. PDN blocks PBL growth in the G1 phase of cell cycle and inhibits both IL-2 receptor (IL-2R) expression and IL-2 secretion. Apoptosis is clearly increased by PDN in PHA-activated human PBL, and the apoptotic effect of PDN is stronger on CD8+ than on CD4+ T lymphocytes. All these effects are dose- and time-dependent. The addition of exogenous IL-2 did not rescue lymphocytes from PDN-increased apoptosis. These results show that PDN increases apoptosis in mature activated human peripheral blood lymphocytes, suggesting a possible role of GCH in the maintenance of immune tolerance at post-thymic level.
Defective integration of activating signals derived from the T cell receptor (TCR) and costimulatory molecules in both CD4+ and CD8+ T lymphocytes of common variable immunodeficiency (CVID) patients
Authors: THON V , WOLF H M , SASGARY M , LITZMAN J , SAMSTAG A , HAUBER I , LOKAJ J , EIBL M M
Published: August 2010

CVID is characterized by hypogammaglobulinaemia and impaired antibody production. Previous studies demonstrated defects at the T cell level. In the present study the response of purified CD4+ and CD8+ T lymphocytes to stimulation with anti-TCR monoclonal antibody (the first signal) in combination with anti-CD4 or anti-CD8, anti-CD2 and anti-CD28 MoAbs (the costimulatory signals) was investigated. Both CD4+ and CD8+ T cells from the patients showed significantly reduced IL-2 release following stimulation via TCR and costimulation via CD4 or CD8 and CD2, respectively. However, normal IL-2 production following TCR plus phorbol myristate acetate (PMA) costimulation and normal expression of an early activation marker, CD69, after TCR + CD28 stimulation indicated that TCR was able to transduce a signal. Furthermore, both IL-2 and IL-4 release were impaired in CD4+ lymphocytes following TCR + CD28 stimulation. In addition, stimulation via TCR + CD28 resulted in significantly decreased expression of CD40 ligand in the patients. These results suggest that the integration of activating signals derived from the TCR and costimulatory molecules is defective in CVID patients; the defect is not confined to costimulation via a single molecule, or restricted to cells producing Thl-type cytokines such as IL-2, and is expressed in both CD4+ and CD8+T cell subsets.
IgA from HIV+ haemophilic patients triggers intracellular signals coupled to the cholinergic system of the intestine
Authors: SALES M E , STERIN-BORDA L , DE BRACCO M M E , RODRIGUEZ M , NARBAITZ M , BORDA E
Published: August 2010

IgA was obtained from HIV-infected haemophilic patients and the intracellular signals triggered by its reaction with isolated rat intestinal strips were studied. HTV+ IgA stained intestinal microvilli with a granular immunofluorescence pattern and bound to the muscarinic acetylcholine receptor (mAChR), displacing the specific muscarinic cholinergic antagonist QNB in a non-competitive manner. It triggered the signals that are the consequence of mAChR stimulation in die intestine. Thus, it decreased cAMP synthesis and increased guanosine 3′:5′-cyclic monophosphate (cGMP) formation and phosphoinositide (PI) turnover of the intestine. In addition, it stimulated prostaglandin E2 (PGE2) synthesis by intestinal strips. Through its effect on PGE2 synthesis, HIV+ IgA could have a dual action. On the one hand, it could enhance immunosuppression at a local level, favouring pathogen growth and subsequent intestinal dysfunction. On the other hand, PGE2 could directly increase intestinal motility and electrolyte/fluid loss. Both effects could be involved in intestinal damage in AIDS.
Circulating antibodies against nicotinic acetylcholine receptors in chagasic patients
Authors: GOIN J C , VENERA G , DE JIMÉNEZ BONINO M BISCOGLIO , STERIN-BORDA L
Published: August 2010

Human and experimental Chagas' disease causes peripheral nervous system damage involving neuromuscular transmission alterations at the neuromuscular junction. Additionally, autoantibodies directed to peripheral nerves and sarcolemmal proteins of skeletal muscle have been described. In this work, we analyse the ability of serum immunoglobulin factors associated with human chagasic infection to bind the affinity-purified nicotinic acetylcholine receptor (nAChR) from electric organs of Discopyge tschudii and to identify the receptor subunits involved in the interaction. The frequency of serum anti-nAChR reactivity assayed by dot-blot was higher in seropositive chagasic patients than in uninfected subjects. Purified IgG obtained from chagasic patients immunoprecipitated a significantly higher fraction of the solubilized nAChR than normal IgG. Furthermore, immunoblotting assays indicated that α and β are the main subunits involved in the interaction. Chagasic IgG was able to inhibit the binding of α-bungarotoxin to the receptor in a concentration-dependent manner, confirming the contribution of the α-subunit in the autoantibody-receptor interaction. The presence of anti-nAChR antibodies was detected in 73% of chagasic patients with impairment of neuromuscular transmission in conventional electromyographical studies, indicating a strong association between seropositive reactivity against nAChR and electromyographical abnormalities in chagasic patients. The chronic binding of these autoantibodies to the nAChR could induce a decrease in the population of functional nAChRs at the neuromuscular junction and consequently contribute to the electrophysiological neuromuscular alterations described in the course of chronic Chagas' disease.
Lung surfactant proteins A and D can inhibit specific IgE binding to the allergens of Aspergillus fumigatus and block allergen-induced histamine release from human basophils
Authors: MADAN T , KISHORE U , SHAH A , EGGLETON P , STRONG P , WANG J Y , AGGRAWAL S S , SARMA P U , REID K B M
Published: August 2010

Aspergillus fumigatus is an opportunistic fungal pathogen which, in the immunocompetent host, causes allergic disorders such as allergic rhinitis, allergic sinusitis, hypersensitivity pneumonitis, and allergic bronchopulmonary Aspergillosis (ABPA). In the present study, the interaction of 3-week culture filtrate (3wcf) allergens and various purified glycosylated and non-glycosylated allergens of A. fumigatus with lung surfactant proteins, SP-A and SP-D, was investigated. Purified SP-A and SP-D, isolated from human bronchoalveolar lavage fluid, bound to the 3wcf allergens and purified allergens, gp55 and gp45, in a carbohydrate-specific and calcium-dependent manner. Both SP-A and SP-D did not bind to deglycosylated allergens, suggesting that the ability of SP-A and SP-D to bind certain allergens is mediated through their carbohydrate recognition domains, interacting with the carbohydrate residues on the allergen. Both SP-A and SP-D could inhibit the ability of allergen-specific IgE from Aspergillosis patients to bind these allergens, suggesting that SP-A and SP-D may be involved in the modulation of allergic sensitization and/or development of allergic reactions. The view that SP-A and SP-D play a protective role against airborne allergens is further supported by the demonstration of their ability to inhibit A. fumigatus allergen-induced histamine release from allergic patients' basophils.
The relationship between colonization and haemagglutination inhibiting and B cell epitopes of Porphyromonas gingivalis
Authors: KELLY C G , BOOTH V , KENDAL H , SLANEY J M , CURTIS M A , LEHNER T
Published: August 2010

Passive immunization with the monoclonal antibody 61BG1.3 selectively prevents colonization by Porphyromonas gingivalis in humans (Booth V, Ashley FP, Lehner T. Infect Immun 1996; 64:422–7). The protective MoAb recognizes the j3 component of the RI protease of P. gingivalis which is formed by proteolytic processing of a polyprotein precursor termed PrpRl. This subunit is both a haemagglutinin and an antigen which is recognized by sera from patients with periodontitis. In this study the relationship was investigated between a colonization epitope which is recognized by the MoAb 61BG1.3, a haemagglutinating and B cell epitope which are recognized by sera from patients with periodontitis. B cell epitopes were mapped by Western blotting with a series of truncated recombinant polypeptides spanning the adhesion domain within residues 784–1130 of PrpRl and by ELISA using a panel of synthetic peptides spanning the same sequence. The epitope which is recognized by the protective MoAb was mapped within residues 907–931 of PrpRl, while serum responses of patients were directed predominantly to the adjacent carboxy-terminal sequence within residues 934–1042. The haemagglutinating epitope was mapped to residues 1073–1112. In view of our previous findings that the MoAb 61BG1.3 prevents colonization of P. gingivalis in vivo and inhibits haemagglutination, these two epitopes may be in proximity in the native protein. Active or passive immunization strategies which target the protective or haemagglutinating epitopes of the adhesion domain of PrpRl may provide a means of preventing infection with P. gingivalis.
Colon carcinoma cell lines stimulate monocytes and lamina propria mononuclear cells to produce IL-10
Authors: KUCHARZIK T , LÜGERING N , WINDE G , DOMSCHKE W , STOLL R
Published: August 2010

Cytokines released from tumour cells may have function as signals to neighbouring immune and inflammatory cells. Several studies have shown that the immunoregulatory cytokines IL-10 and transforming growth factor-beta 1 (TGF-β1) as well as prostaglandin-E2 (PGE2) play an important role in tumour-induced immunosuppression. The aim of the study was to investigate the effect of colon carcinoma cell lines on IL-10 production in peripheral monocytes (PBMC) and lamina propria mononuclear cells (LPMC). We examined four colon carcinoma cell lines (HT-29, Caco-2, Colo-320 and HCT-116) and determined their production of TGF-β1, IL-10 and PGE2. Peripheral monocytes were isolated by density gradient centrifugation and LPMC were isolated from surgical specimens using a collagenase digestion method. Monocytes and LPMC were cultured with colon carcinoma cell conditioned medium or in co-culture with colon carcinoma cells. Supernatants were then determined for the production of IL-10 by ELISA assays. All colon carcinoma cell lines stimulated peripheral monocytes as well as LPMC to produce markedly increased levels of IL-10. Colon cancer cells secreted negligible levels of IL-10, but high amounts of TGF-β1 and PGE2. Neutralization of TGF-β1 by administration of anti-TGF-β as well as neutralization of PGE2 with anti-PGE2 antisera reduced the IL-10 production of monocytes markedly, indicating that tumour cell-derived TGF-β1 and PGE2 are major factors for IL-10 stimulation. In vitro stimulation of monocytes with TGF-β1 and PGE2 could confirm that TGF-β1 as well as PGF2 at picogram concentrations were able to prime monocytes for enhanced IL-10 production. Our results demonstrate that colon carcinoma cell lines enhance the ability of monocytes and intestinal macrophages to produce IL-10. The stimulation of monocyte IL-10 by colon cancer cell-derived TGF-β1 and PGE2 may act as a tumour-protecting mechanism by impairing the activation of anti-tumour cytokines.