bluuebunny/crossref_metadata_2025 huggingface dataset:
Immune complex-degradation ability of macrophages in MKL/Mp-lpr/lprlupus mice and its regulation by cytokines
Authors: KANNO H , TACHIWAKI O , NOSE M , KYOGOKU M
Published: July 2010

Impaired clearance of circulating and/or deposited immune complexes (IC) by the mononuclear phagocylic system is one of the major factors in the pathogenesis of IC diseases. MRL/Mp-lpr/lpr (MRL/lpr) lupus mice spontaneously develop a lethal glomerulonephritis associated with IC deposition. The ability of macrophages to degrade phagocytozed IC and regulation of this degradation in MRL/lpr mice were examined. In 4-month-old MRL/lpr mice. Macrophages accumulated in the affected glomeruli and these macrophages contained many phagosomes containing electron-dense bodies. When culture supernatant of human T cell line HUT102 was administered intraperitoneally into disease-bearing MRL/lpr- mice, degradation of these electrondense bodies in the macrophages in glomeruli was noted. We developed a quantitative in vitro assay for IC degradation activity of MRL/lpr resident peritoneal macrophages (RPM) using peroxidase-labelled IC derived from MRL/lpr mouse sera. The ability of the RPM to degrade IC was remarkably enhanced by the pretreatment with HUT102 cell products and the related human recombinant cytokines lymphotoxin and IL-1α. Moreover, pretreatment of RPM from non-diseased MRL/Mp- +/+ mice with the culture supernatant of spleen cells from diseased MRL/lpr mice reduced their IC degradation activity. These results suggested that the ability of macrophages to degrade IC in MRL/Mp strains of mice is under the regulation of cytokines and the impaired ability in the disease-bearing mice may be the result of abnormalities in the cytokine system in these mice.
Regulation of human basophil activation; the role of Na+ and Ca2+in IL-3-induced potentiation of IgE-mediated histamine release from human basophils
Authors: BEAUVAIS F , ECHASSERIEAU K , BURTIN C , BENVENISTE J
Published: July 2010

The release of mediators from human basophils is strongly enhanced by lL-3. However, the signalling pathways of IL-3 arc poorly defined in these cells. Since external Ca2+ and Na+ play important regulating roles in histamine release, the possibility that these cations could be involved in the potentiation by rL-3 of the anti-IgE-induced histamine release from human basophils was considered, and it was observed that: (i) lL-3 dramatically decreased the external Ca2+ requirement for IgE-mediated histamine release. However, histamine release from IL-3-treated basophils became only partially independent of external Ca2+, since addition of EGTA in the external medium abolished the effect of IL-3; (ii) decreasing Na+ influx by lowering external Na+ concentration in isosmotic medium inhibited thepotenEiatiiigefTeLtoriL-3on IgE-mediated release; (iii) amiloride. An inhibitor of Na+/Ca2+ and Na+/H+ exchanges, and its derivative, benzamil, more specific for Na+/Ca2+ exchanges, inhibited the release potentiated by IL-3. In contrast, the amiloride derivative 5-(N, N-dimethy1)-amiloride more specific for Na+/H+ exchanges, slightly increased the IL-3-enhaneed release. Thus, the decreased requirement for external Ca2+ and the dependence on external Na + taken with the effect of the Na+/Ca2+ exchange inhibitors, suggest that Na+/Ca2+ exchanges are involved in the IL-3-indueed enhancement of IgE-mediated human basophil histamine release.
Dichotomy of the human T cell response toLeishmaniaantigens. I. Th1-like response toLeishmania majorpromastigote antigens in individuals recovered from cutaneous leishmaniasis
Authors: KEMP M , HEY A S , KURTZHALS J A L , RISTENSEN C B V C H , GAAFAR A , MUSTAFA M D , KORDOFANI A A Y , ISMAIL A , KHARAZMI A , THEANDER T G
Published: July 2010

The T cell response to antigens from Leishmania major promastigotes was investigated in peripheral blood mononuclear cells from Sudanese individuals with a history of cutaneous leishmaniasis (CL), Sudanese individuals with positive DTH reaction in the leishmanin skin test but with no history of skin lesions, and in Danes without known exposure to Leishmania parasites. Proliferation and production of interferon-gamma (IFN-γ) and IL-4 in antigen-stimulated cultures was measured. Lymphocytes from individuals with a history of CL proliferated vigorously and produced IFN-γ after stimulation with either a crude preparation of L. major antigens or the major surface protease gp63. These cultures produced no or only little IL-4. Also cells from leishmanin skin test-positive donors with no history of CL produced IFN-γ and no IL-4 in response to major antigens. Cells from the unexposed Danes were not activated by gp63. The cells from Danish donors produced either IFN-γ or IL-4, but not both cytokines after incubation with the crude preparation of L. major antigens. The data show that the T cell response to Leishmania antigens in humans who have had uncomplicated CL or subclinical L. major infection is an IFN-γ-producing Th1-like response.
Augmentation of antigen-specific lymphoproliferative responsesin vitroby biological response modifiers
Authors: GRUIJL T D D E , MOORE J J , DE VRIES E , VON BLOMBERG-VAN DER FLIER B M E , FONK J C M , SCHEPER R J
Published: July 2010

The detection of antigen-specific T cell responsiveness, particularly of resting memory lymphocytes, in cultures of peripheral blood mononuclear cells (PBMC) may be hampered by a less than optimal antigen presentation in vitro. Augmented sensitivity of the test system may be achieved by the addition of reagents with a beneficial effect on lymphocyte and antigen-presenting cell (APC) functions. In this study the effect of several biological response modifiers on antigen-specific T cell proliferation was determined, using nickel sulphate and tetanus toxoid as lest antigens. IL-lα (100 U/ml). interferon-gamma (IFN-γ) (10 U/ml), and indomethacin (2 μM) were found to significantly enhance nickel-induced proliferation in PBMC cultures from nickel-hypersensitive donors (n = 6). Tetanus-induced proliferation (n = 5) was similarly enhanced, both by the above supplements and by the addition of polyethylene glycol (PEG) or a neuraminidase treatment of the PBMC before culture. The addition to PBMC cultures of a combination of IL-lα (30 U/ml), IFN-γ (10 U/ml), and indomethacin (2 μM) is recommended to specifically enhance antigen-induced lymphoproliferative signals.
Pathogenic natural anti-cardiolipin antibodies: the experience from monoclonal gammopathy
Authors: COHEN J , BAKIMER R , BLANK M , VALESINI G , SHOENFELD Y
Published: July 2010

Anti-cardiolipin antibodies (ACA) were detected in 19% of sera from patients with monoclonal gammopathies (MG). ACA were purified from the sera of patients with MG. One of the IgG-ACA was found to be monospecific with high affinity for cardiolipin, and to carry a pathogenic ACA Id (1.10), Active immunization of naive BALB/c mice with the puri6ed IgG-ACA was followed by production in the mice of sustained high titres of ACA, associated with prolonged activated partial thromboplastin time (APTT) (61 ± 14s versus 31 ± 2 s in control mice; P < 0.001) and thrombocytopenia (468 000 ± 224 000/mm3 versus 994 000 ± 92 000/mm3 in controls; P < 0001). The titres of other autoantibodies (e.g. anti-DNA, anti-histones), although being high after immunization, decreased rapidly and were undetected after 1 month following the boost injection. The mice immunized with the IgG-ACA exhibited low fecundity (36% of mice became pregnant versus 62% observed in the group immunized with control IgG). The pregnant mice had increased resorption rate (the equivalent of fetal loss in the human) of 52 ± 8% (versus 5 ± 4% in the control group). The mean (±s.d.) embryo and placental weights in mice with anti-phospholipid syndrome (APLS) were significantly lower compared with the mice injected with control IgG (682 ± 304 mg and 102±12mg versus 1303 ± 105 mg and 145±8mg, respectively; P<0.001). Serum monoclonal immunoglobulins having autoantibody activity may be regarded as an expansion of clones producing natural autoantibodies. Our results confirm the pathogenic role of natural ACA in the pathogenesis of the anti-phospholipid syndrome.
Persistence of a rheumatoid factor (RF)-producing B cell clone with a somatically mutated Igκ chain in a patient with rheumatoid arthritis
Authors: SCHRAUDER A , GAUSE A , JUNG W , MIERAU R , PFREUNDSCHUH M
Published: July 2010

The VκIV gene encoding the light chain of an IgA RF has been shown to have undergone 31 somatic mutations compared with the single existing VκIV germ-line gene. We now show the persistence of the rearranged and mutated DNA coding for this RF over a period of 5 years in the peripheral blood lymphocytes (PBL) of the patient with rheumatoid arthritis (RA). The sequence of the RF has been conserved to identity over this period. These results raise the possibility that the particular antigenic stimulus leading to RF production in this RA patient is active over a long period of time.
Enhanced generation of reactive oxygen species in monocytes from patients with common variable immunodeficiency
Authors: AUKKUST P , MÜLLER F , FRØLAND S S
Published: July 2010

Monocyte and macrophage dysfunction may be important for both immunopathogenesis and clinical manifestations in subgroups of patients with primary hypogammaglobulinaemia. In the present study we examined the ability to generate reactive oxygen species (ROS) in isolated monocytes from these patients by two different methods: superoxide dismutase (SOD) inhibitable cytochrome c reduction by O2− and nitroblue tetrazolium (NBT) reduction. Monocytes from patients with common variable immunodeficiency (CVI) demonstrated significantly enhanced ROS generation both unstimulated and stimulated (unopsonized zymosan and phorbol myristate acetate (PMA)). The enhanced oxidative burst response in CVI patients was found both with and without scrum containing medium. Furthermore, serum from CVI patients did significantly enhance the oxidative burst response in monocytes from healthy blood donors compared with the effect of control serum. The enhanced ROS generation in CVI patients was significantly correlated with elevated serum levels of neopterin, reduced numbers of CD4+ lymphocytes in peripheral blood and occurrence of splenomegaly. In contrast to the CVI group, monocytes from patients with X-linked agammaglobulinaemia (XLA) did not show enhanced ROS generation. The increased oxidative burst response in monocytes from CVI patients most probably reflects in vivo activation of these cells. Our observations indicate the presence of a subgroup of CVI patients characterized by chronic immune activation particularly of monocytes. The enhanced ROS generation might be involved in immunopathogenesis (e.g. T cell dysfunction) and in the pathogenesis of clinical manifestations (e.g. malignancies and autoimmune disorders) in these patients.
Serum soluble CD23 in patients with hypogammaglobulinaemia
Authors: BANSAL A S , HAENEY M R , COCHRANE S , PUMPHREY R S H , GREEN L M , BHAVNANI M , WILSON P B
Published: July 2010

Serum levels of the soluble form of the low-affinity receptor for IgE (FcERII, CD23) (sCD23) are elevated in autoimmune conditions associated with hypergammaglobulinaemia and B cell hyperactivity. Very high levels of sCD23 are found in patients with B-chronic lymphatic leukaemia (B-CLL) who are, however, frequently hypogammaglobulinaemic. We therefore compared the serum levels of sCD23 in healthy controls (n= 33) with three conditions associated with hypogammaglobulinaemia (HGG) and varying B cell numbers: X-linked agammaglobulinaemia (XLA, n= 12), common variable immunodeficiency (CVI, n= 20) and B-chronic lymphatic leukaemia (n= 33). Serum levels of sCD23 showed a significant correlation with the CD19+ B cell count in both normals and patients with CVI (r= 0 65, P < 0·0001). Amongst the different clinical groups, serum levels of sCD23 were increased in the order XLA < CVI < normals < CLL (medians 2·5, 7·7, 11·1 and 540, respectively; P < 0·001 for all comparisons except CVI versus normals P < 0·03 in a one-tailed test). In the CVI group, serum sCD23 was lowest amongst four patients with low B cell numbers. There was no overlap in sCD23 between patients with XLA and this subgroup of CVI patients. Serum sCD23 is, therefore, derived predominantly from B cells, and is significantly related to the peripheral blood B cell count.
T cell receptor repertoire and mitotic responses of lamina propria T lymphocytes in inflammatory bowel disease
Authors: QIAO L , GOLLING M , AUTSCHBACH F , SCHURMANN G , MEUER S C
Published: July 2010

Human intestinal lamina propria T lymphocytes (LPL-T) physiologically exhibit minimal proliferation in response to antigen receptor stimulation in vitro. This is thought to occur as a consequence of regulatory influences which are exerted by the mucosal microenvironment. The present study is aimed at investigating whether proliferative responses of intestinal LPL-T to antigen receptor stimulation are altered in patients with inflammatory bowel disease. Accordingly, proliferative responses of LPL-T in patients with Crohn's disease and ulcerative colitis to stimulation with CD3 MoAb plus IL-2 were examined and compared with controls. In addition, T cell receptor (TCR) repertoires of LPL-T and peripheral blood T lymphocytes were determined by indirect immunofluorescenee using a panel of 11 TCR Vβ specific antibodies. In most patients with inflammatory bowel disease, LPL-T showed enhanced proliferation to antigen receptor stimulation compared with controls. Moreover, perhaps as a consequence, an enhanced frequency of in vivo preactivated T cells was seen as judged from an increased spontaneous proliferative response to low concentrations of exogenous IL-2. LPL-T and peripheral blood T lymphocytes exhibited similar percentages of TCR Vβ gene usage both in controls and in patients. In summary, polyclonal activation of LPL-T due to impairment of local adjustment, i.e. insufficient down-regulation of TCR/CD3-dependent signalling processes, may contribute to the pathogenesis of inflammatory bowel disease.
Combined effect of fluconazole and thymosin α1 on systemic candidiasis in mice immunosuppressed by morphine treatments
Authors: DI FRANCESCO P , GAZIANO R , CASALINUOVO I A , BELOGI L , PALAMARA A T , FAVALLI C , GARACI F
Published: July 2010

Treatment of systemic infection with Candida albicans with a combination of an antifungal agent (i.e. fluconazole) and a thymus-derived immunostimulant (i.e. thymosin α1 (Tα1)) in mice immunosuppressed by morphine treatments was investigated. In normal mice, fluconazole given after infection with 106  C. albicans cells was more effective than in mice treated with morphine. Combination treatment with fluconazole and Tal prolonged survival and reduced the fungal burden in the kidneys of immunosuppressed mice. We also investigated the influence of this combined treatment on killing properties of polymorphonuclear leucocytes (PMN) and natural killer (NK) cell activity, inhibited by morphine administrations. Treatment with Tα1 or fluconazole as single agents promoted a recovery of normal NK cell activity and intracellular killing of C. albicans by PMN, while the combination significantly increased both of these responses, probably through the modulation of lymphokine production. Our data suggest that the additive effect of Tα1 and fluconazole is due lo a direct antifungal action and activation of the immunocompetence.
Levels of soluble VCAM-1, soluble ICAM-1, and soluble E-selectin during disease exacerbations in patients with systemic lupus erythematosus (SLE); a long term prospective study
Authors: SPRONK P E , BOOTSMA H , HUITEMA M G , LIMBURG P C , KALLENBERG C G M
Published: July 2010

Active SLE is characterized by immune deposits and subsequent vascular inflammation in many organs. Expression and up-regulation of adhesion molecules is basic to migration of inflammatory cells into the tissues. Recently, soluble isoforms of these molecules have been described which might be an expression of their up-regulation in the tissues and, as such, of disease activity. The purpose of this study was to evaluate whether changes in levels of soluble adhesion molecules reflect disease activity. We analysed serial sera in a 6-month period preceding 22 consecutive exacerbations of SLE for levels of soluble vascular cell adhesion molecule-l (sVCAM-1). soluble intercellular adhesion molecule-l (slCAM-1), and sE-selectin. Levels were related to clinical disease activity (SLEDAI). and levels of anti-dsDNA and complement. At the time of maximal disease activity, levels of sVCAM-1 in patients with SLE were higher than those in controls (P < 0.0001), levels in patients with renal involvement being higher than in those without (P < 0.02). Levels of sVCAM-1 correlated with SLEDAI scores (P < 005) and, inversely, with levels of C3 (P = 0.01). In addition, in the presence of anti-dsDNA, levels of sVCAM-1 tended to correlate with levels of these autoantibodies (P < 0.1). Levels of sICAM-1 were normal and sE-selectin levels even decreased compared with controls. Levels of sVCAM-l were higher at the moment of relapse (P = 0.001) than at 6 months before this time point. This rise correlated with the rise in SLEDAI score (P < 0.02). Levels of sICAM-l and sE-selectin did not rise, and remained in the normal range in all exacerbations studied. In conclusion, in contrast to sICAM-1 and sE-selectin, levels of sVCAM-l are increased, rise parallel to disease activity during exacerbations in SLE, and are associated with decreasing levels of complement factors. This favours the hypothesis of immune deposit formation, activation of the complement cascade and activation of endothelial cells. Concurrent up-regulation of vascular adhesion molecules may thus result in transmigration of activated inflammatory cells inducing tissue damage.
Cytotoxic effects of antibodies to proteinase 3 (C-ANCA) on human endothelial cells
Authors: MAYET W-J , SCHWARTING A , MEYER ZUM BÜSCHENFELDE K-H
Published: July 2010

Autoantibodies directed against cytoplasmic antigens of neutrophils (ANCA), especially those with specificity for proteinase 3 (PR-3) and myeloperoxidase, are valuable markers for differential diagnosis and monitoring of disease activity in Wegener's granulomatosis (WG) and other vasculitides. Till now, several concepts concerning a direct role of antibodies against PR-3 in the pathogenesis of WG have been discussed. Recently we were able to show that these antibodies recognize PR-3 translocated into the membrane of human endothelial cells. The aim of this study was to investigate putative cytotoxic effects of antibodies to PR-3 on human endothelial cells. Antibodies were obtained by affinity purification of sera from patients with active WG, Purified antibodies to Ro (SS-A), La (SS-B) and RNP served as controls. Purified antibodies to PR-3 displayed a lytic activity against endothelial cells treated with tumour necrosis factor-alpha (TNF-α) with the help of cytokine-primed neutrophils as measured in a Cr-release assay. About 100% specific cytotoxicity occurred after 4 h and was independent of complement, Cytotoxic effects were inhibited by coincubation with unprimed ncutrophils or preincubation of PR-3 antibodies with purified antigen. Antibodies to Ro (SS-A), La (SS-B) or RNP had no cytotoxic effect. In summary, PR-3 antibody-induced cytotoxicity required (i) expression of PR-3 on the surface of TNF-α-treated endothelial cells: and (ii) co-cultivation of cytokine-primed neutrophils. This is to the best of our knowledge the first report on direct cytotoxic effects of PR-3 antibodies on vascular endothelium. Our data give a hint at a PR-3 antibody-mediated mechanism of endothelial injury via antibody-dependent cellular cytotoxicity in WG and other ANCA-related vasculitides.
Reduced interferon-gamma (IFN-γ) secretion with increased IFN-γ mRNA expression in atopic dermatitis: evidence for a post-transcriptional defect
Authors: TANG M L K , VARIGOS G , KEMP A S
Published: July 2010

Reduced secretion of IFN-γ in atopic individuals has been implicated in the pathogenesis of disease, though the mechanisms leading to this reduced secretion have not been elucidated. As production of IFN-γ has been shown to be predominantly regulated by its rate of transcription, expression of IFN-γ mRNA was examined in atopic children and in age-matched, non-atopic controls by polymerase chain reaction (PCR)-assisted mRNA amplification. Children with atopic dermatitis were found to have constitutive expression of IFN-γ mRNA in freshly isolated peripheral blood mononuclear cells (PBMC) and in unstimulated PBMC cultures which increased further following stimulation with phorbol myristate acetate (PMA)/Ca in vitro. In contrast, expression of IFN-γ mRNA in controls was only detected in stimulated cultures, as has been demonstrated previously for normal adults. These findings demonstrate that circulating T cells from atopic children have been activated in vivo, and suggest that T cell activation is a significant component of the inflammatory process in atopic dermatitis. Although expression of IFN-γ mRNA was increased in the atopic children, secretion was confirmed to be significantly lower than in controls, indicating that the defect(s) underlying reduced IFN-γ secretion in these individuals lie post-transcriptionally.
Analysis of lymphocyte cell death and apoptosis in HIV-2-infected patients
Authors: JALECO A C , COVAS M J , VICTORINO R M M
Published: July 2010

Recent evidence suggests that T cell apoptosis could be involved in the pathogenesis of HIV-1 infection. As the progression of HIV-2 associated disease appears to be slower than that of HIV-1, we investigated whether there were differences in the degree of T cell death and apoptosis in peripheral blood mononuclear cell (PBMC) cultures from patients with HIV-1 or HIV-2 infection. PBMC from healthy controls (n = 28) and patients infected with HIV-1 (n = 26: asymptomatic (ASY)/persistent generalized lymphadenopathy (PGL). w = 16; and AlDS-related complex (ARC)AIDS n = 10) or HIV-2 (n = 30: ASY/PGL, n = 16: ARC/ AIDS, n = 14) were cultured in the absence or presence of mitogens (PHA, PWM) or superantigen (SEB). After 48 h, cell death (CD) was assessed by trypan blue exclusion and in some patients programmed cell death (PCD) was quantified in flow cytometry by measuring the percentage of hypodiploid nuclei corresponding to fragmented DNA, after treating the cells with a propidium iodide hypotonic solution. HlV-1 and HlV-2 ARC/AIDS patients and ASY/PGL HIV-1+ patients had significant increases in cell death percentages compared with controls, both in unstimulated and stimulated lymphocyte cultures. However, HIV-2+ ASY/FGL patients did not exhibit significant increases of cell death in unstimutated cultures. In addition, the comparison between HIV-l and HIV-2 infected subjects in similar stages of disease, showed no significant differences in CD in the ARC AIDS patients, although ASY/PGL HIV-2-infected subjects had lower levels of CD than the HIV-1+ ASY/PGL (3.4%± 0.6 s.e.m. versus 6.8%± M s.e.m., P < 0.01). PCD was significantly increased both in ASY/PGL (14.3%± 2.2 s.e.m., n = 8, P< 0.005) and m ARC/AIDS (25.3%± 4.5 s.e.m., n = 9, P < 0.001) HIV-1+ patients compared with healthy controls (5.8%± l.7 s.e.m., n = 11). This contrasts with HIV-2 infected subjects where the ASY/PGL patients (10.0%± 2.8 s.e.m., n = 6) did not differ significantly from healthy controls, although ARC/AIDS patients (27.2%± 4.2 s.e.m., n = 9. P < 0.001) had significantly increased levels of PCD. In conclusion, this is the first report describing the occurrence of spontaneous and activation-induced lymphocyte death by apoptosis in HlV-2 infected subjects. The lower levels of PCD in ASY/PGL HlV-2 infected patients compared with HIV-T patients at a similar stage justify further investigation to define whether these differences have any rote in the putative slower progression of HlV-2 disease.
Pneumocystis carinii-induced activation of the respiratory burst in human monocytes and macrophages
Authors: LAURSEN A L , MØLLER B , RUNGBY J , PETERSEN C M , ANDERSEN P L
Published: July 2010

Human monocytes and monocyte-derived macrophages were studied for their ability to phagocytose Pneumocystis carinii and produce superoxide (O−2) during the process. One × 106 freshly isolated monocytes. incubated with 0.1 – 3.75 × 106P. carinii cysts, increased O−2 production in a dose-related way. Antibodies were essential for the process since opsonized, but not unopsonized, pneumocysts induced O−2 production significantly above the response obtained by lung tissue from rats (10.7 and 4.9 versus 30 fmol/cell per 90 min). The difference between pneumocysts opsonized in untreated versus complement-depleted serum was not significant (10.7 versus 12.6 fmol/cell per 90 min). Monocyte-derived macrophages also activated the respiratory burst when stimulated with pneumocysts, and this effect could be significantly increased, from 4.2 to 8.8 fmol/cell per 90 min, when cells were primed with interferon-gamma (IEN-γ). Cells primed with IL-3 also increased O2 production, though to a lesser extent. In contrast, granulocyte-macrophage colony-stimulating factor (GM-CSF) had only a small effect on the respiratory burst in cells stimulated with P. carinii. Priming with IEN-γ increased the rate of phagocytosis in macrophages. After incubation for 90 min or more, however, the percentage of cells with phagocytic vacuoles was only slightly higher in IFN-γ-primed cells. When examined by electron microscopy (EM), most vacuoles contained partially or totally degraded pneumocysts. In conclusion, we have demonstrated the ability of monocytes and monocyte-derived macrophages to ingest and degrade pneumocysts. activating the respiratory burst during the process.
Induction in vitro of 72-kD heat shock protein in a continuous culture of rat thyroid cells, FRTL5
Authors: MISAKI T , TAKEUCHI R , MIYAMOTO S , HIRANO A , KASAGI K , KONISHI J
Published: July 2010

In Graves' disease and Hashimoto's thyroiditis, the presence of 72-kD heat shock protein (hsp-72) on thyrocytes has been reported. To clarify the significance of this phenomenon, we induced the antigen in thyroid cell culture in vitro. In the FRTL5 rat cell line, which had been healed at 425 C or treated with sodium arsenite, expression of hsp-72 was examined with immunoperoxidase staining and immunoprecipitation of the metabolically labelled protein using a specific MoAb. In the cells cultured either with or without thyrotropin (TSH), heat and chemical stresses reproducibly and dose-dependently induced hsp-72 antigen, whereas unstimulated controls had no significant immunoreactivity. Unlike in Graves' retroocular fibroblasts, hydrogen peroxide was not an effective stress in FRTL5, and the induction was not suppressed by methylmercaptoimmidazole and propylthiouracil, nor enhanced by interferon-gamma (IFN-γ) and tumour necrosis factor-alpha (TNF-α), These data could not support the hypotheses that suppression of thyroid autoimmunity by thionamides is due to their modulatory action on hsp-72 expression, or that presence of that antigen in the thyroid tissues affected by autoimmunity is secondary to cytokine secretion from infiltrating immunocytes. On the other hand, coculture experiments of stressed FRTL5 cells and syngeneic Fisher rat splenocytes suggest that aberrantly expressed hsp may activate part of the thyroid-infiltrating lymphocytes and thereby aggravate autoimmune processes. The induction and detection systems of hsp-72 using FRTL5 cells would facilitate future studies, possibly utilizing human materials as well, to explore possible relations between stress proteins and thyroid autoimmunity.
Podocyte expression of MHC class I and II and intercellular adhesion molecule-1 (ICAM-1) in experimental pauci-immune crescentic glomerulonephritis
Authors: COERS W , BROUWER L , VOS J T W M , CHAND A , HUITEMA S , HEERINGA P , KALLENBERG C G M , WEENING J J
Published: July 2010

We examined immunopathological changes of podocytes in vivo which, based on in vitro studies, are thought to be relevant for the pathogenesis of renal diseases. We investigated the alterations of podocytes in local inflammation in a recently developed model of pauci-immune necrotizing crescentic glomerulonephritis (NCGN) in the rat. Frozen and plastic embedded kidney sections at different time points of the disease were incubated with antibodies directed to MHC class I, MHC class II, ICAM-1 and to relevant cytokines. Strong glomerular expression of MHC class I. II and ICAM-1 was found within 4 days, and plastic embedded sections clearly demonstrated increased cell membrane staining of podocytes. Increased glomerular interferon-gamma (IFN-γ) was detected within 24 h of induction of NCGN. and IL-IβT and tumour necrosis factor-alpha (TNF-α) were found from day 4. The potency of these cytokines to induce adhesion molecules on podocytes was investigated on rat glomerular epithelial cells in vitro. By using IACS analysis and electron microscopical techniques, we found that the in vivo expression of MHC class I, II and ICAM-I by podocytes could in vivo be simulated by IFN-γ IFN-α weakly induced MHC class I. while IL-Iβ and TNF-α were ineffective. We hypothesize that podocytes in this in vivo model are important to maintain the local inflammatory process in the glomerulus by expression of relevant adhesion molecules and MHC molecules upon stimulation with specific cytokines.
Influence of an established acute phase response on the severity of experimental nephritis
Authors: KARKAR A M , REES A J
Published: July 2010

Small doses of lipopolysaccharide (LPS) induce an acute phase response (APR), and a number of studies have also shown that this greatly enhances the severity of glomerular injury in the heterologous phase of nephrotoxic nephritis (hNTN), an experimental model of anuglomerular basement membrane (GBM) disease. Here, we examined the influence of pre-existing subclinical infection and raised APR assessed by plasma α2-macrogiobulin (α2-M) concentration, on the degree of injury in this model of nephritis. Studies were initially performed to determine the normal range of α2-M in rats and its modulation by IL-6 and different doses of LPS. Plasma concentration of α2-M was found to be variable and dependent on the weight of the rats. Single injections of either LPS or IL-6 had a comparable effect and continuous perfusions of LPS caused a progressive increase in α2-M which peaked al 4S h and declined gradually over 1 week. Following induction of nephritis with long of anti-GBM antibody, rats with raised α2-M had 14 ± 3 mg 24 h albuminuria compared with 4 ± 1 mg 24 h in rats with normal α2-M (P < 0.0.001, Wilcoxon). Injection of 20 mg anti-GBM antibody caused 36 ± 11 mg 24 h albuminuria compared with 16 ± 4 mg/24h (P < 0001), respectively. However, all these rats remained active and none of them died, hi contrast, injection of 0.25 μg LPS before induction of nephritis with 10 mg anti-(IBM antibody, in rats with raised a,-M, caused severe albuminuria (115 ± 23 mg 24 h) compared wilh rats having normal levels of α2-M (72 ± 15 mg 24h. P < 005), Furthermore, rats with raised α2-M also had severe systemic manifestations characterized by pulmonary haemorrhage and extensive glomerular thrombosis, and many of them died. These results demonstrate the potential effect of pre-existing subclinical infection and raised APR on severity of glomerular injury which may affect the outcome of experimental studies.
Reactive oxygen product formation by human neutrophils as an early marker for biocompatibility of dialysis membranes
Authors: ROSENKRANZ A R , TEMPL E , TRAINDL O , HEINZL H , ZLABINGER G J
Published: July 2010

Production of reactive oxygen intermediates (ROI) by neutrophils (PMN) in vivo was examined by a whole blood assay using dichlorofluorescein-diacetate (DCFH-DA) in 10 patients each dialyser consecutively with two different dialyser membranes. Haemodialysis (HD) with cuprophan membrane (CM) led to a significantly (P < 0001) more pronounced ROI production by PMN (2.4 ± 0.5-fold increase in Intracellular oxidation of DCFH-DA) compared with HD with polysulfone membranes (PM; 1.5 ± 0.2-fold). HD with CM induced a decrease in PMN count by about 90%, whereas PM induced a decrease by only 25% (P < 0.001). In CM patients maximal ROI production coincided with the nadir in PMN count. All patients dialysed with CM showed a clear increase in serum levels of Bb fragments. whereas PM-dialysed patients did not. In this respect however, no clear time relationship was seen to the kinetics of ROI production, nor to the disappearance of neutrophils from the circulation valuating a direct effect of the dialysis membranes on PMN demonstrated that incubation of neutrophils with hollow fibres of the CM but not of the PM in the absence of plasma induces significant ROI production by PMN. Our study thus indicates that ROI production by PMN during HD correlates to membrane biocompatibility. Furthermore, one might speculate that also independently from but perhaps in addition to complement activation, reactive oxygen products arc critically involved in the generation of haemodialysis-associated neutrophil emigration.
Cellular mRNA expression of interferon-gamma, IL-4 and transforming growth factor-beta (TGF-β) by rat mononuclear cells stimulated with peripheral nerve myelin antigens in experimental allergic neuritis
Authors: ZHU J , MIX E , OLSSON T , LINK H
Published: July 2010

Experimental allergic neuritis (EAN) serves as a useful model for inflammation in the peripheral nervous system. To study the potential role of important immunoregulatory and effector cytokines in EAN, we examined the expression of mRNA for interferon-gamma (IFN-γ), IL-4 and JGF-β by in situ hybridization in lymph node and splenic cells cultured with bovine peripheral nerve myelin (BFM), P2 and P0 during the course of EAN in Lewis rats. Levels of IFN-γ mRNA-expressing mononuclear cells (MNC) from lymph nodes and spleens roughly correlated with clinical status, consistent with a disease-promoting role for IFN-γ. BPM, P0 and P2-rcactive IFN-γ mRNA-expressing T cells appeared in lymph nodes and spleen before onset of the disease, whereas a significant TGF-β response to BPM, P2 and PO was observed at lower levels than the IFN-γ response and at onset of recovery, consistent with a disease down-regulating role of TGF-β. IL-4 mRNA-expressing cells were found at levels similar to TGF-β mRNA-expressing cells, and with the latest peak of the three cytokines examined. This result suggests that IL-4 may also suppress IFN-γ expression at late recovery phase of EAN.
Transforming growth factor-beta 1 (TGF-β1)- and β2-like activities in malignant pleural effusions caused by malignant mesothelioma or primary lung cancer
Authors: MAEDA J , UEKI N , OHKAWA T , IWAHASHI N , NAKANO T , HADA T , HIGASHINO K
Published: July 2010

We investigated the levels of TGF-β in malignant pleural effusions (MPE) caused by malignant mesothelioma (MESO) or primary lung cancer. TGF-β levels in MPE caused by MESO were 283.9 ± 219.2pM (mean plusmn; s.d.) and were three to six times higher than those due to primary lung cancers (P < 0.01 or P < 0.05). We also evaluated TGF-β1- and β2-like activities in MPE using specific polyclonal antibodies. Although TGF-β1-like activity could be detected in all cases, TGF-β2-like activities were detected in five of seven in MESO and in a few cases with primary lung cancer. These results demonstrate that the levels of total TGF-β and TGF-β2-like activity may be clinically useful to differentiate MESO from primary lung cancer. Our data also suggest that TGF-β may help further characterize the clinical features of MESO.
Human anti-mouse antibody response to the injection of murine monoclonal antibodies against IL-6
Authors: LEGOUFFE E , LIAUTARD J , GAILLARD J P , ROSSI J-F , WIJDENES J , BATAILLE R , KLEIN B , BROCHIER J
Published: July 2010

We analysed human anti-mouse antibodies (HAMA) in 12 patients (six with multiple myeloma (MM) and six with metastatic renal cell carcinoma (MRCC)) who were treated with B-E8, an IgG1 MoAb against IL-6. Efficiency of the treatment was evidenced by the drop in the serum levels of C-reactive protein (CRP), the in vivo production of which is under the control of IL-6. Three patients with MM and the six patients with MRCC became immunized m the injected MoAb. HAMA appeared between days 7 and 15 after the beginning of the treatment. The nine patients made IgG antibodies: four also made IgM. All immunized patients made anti-biotype antibodies specific to B-E8. Two of them also developed HAMA directed to murine IgG1 isotype: in these two patients B-E8 MoAb cleared rapidly from the circulation with loss of treatment efficiency. In the patients who developed only anti-biotype antibodies, serum levels of B-E8 remained unchanged and CRP production remained inhibited, indicating that treatment remained efficient in the presence of HAMA, Circulating B-E8 MoAbs were still able to bind to iL-6 and to inhibit IL-6-dependent proliferation despite the presence of anti-idiotypic HAMA, Therefore, in contrast to HAMA produced against MoAb directed against cellular targets, HAMA against anti-IL-6 MoAb idiotopes led neither to clearance nor to functional inactivation of the injected MoAb. This was further shown by resuming the B-E8 treatment with success in a patient who still had anti-idiotypic HAMA.
The significance of the pre-challenge immune status of mice for development of retrovirus-induced immunodeficiency syndrome (MAIDS)
Authors: PAVLOVITCH J H , RIZK-RABIN M , PICARD F , MARUSSIG M , HALBREICH A
Published: July 2010

The effects of vaccination with RNA-free viral pseudoparticles, preinfection with non-pathogenic ecotropic virus, and induction of tolerance to viral proteins in newborns on the outcome of murine immunodeficiency syndrome (MAIDS) were studied. The parameters used to follow disease progression were: lymphopenia, circulating B and T8 cells, serum IgG and IgM levels, lymphoproliferation and skin graft rejection, [minimization with RNA-free viral pseudoparticles had no effect on any of these parameters. Preinfection of adults with ecotropic virus and the induction of tolerance in newborns to virus antigens both attenuated the early symptoms of viral infection and delayed the onset of immunodeficiency and lympho-proliferation in some mice, but did not significantly alter the number of deaths due to MAIDS. Failure of immune-based therapy to produce successful protection against MAIDS suggests that immune destruction caused by the persistent virus rather than hyper immune activity is the main pathogenic factor in this disease.
Comparison of the immune response against Mycobacterium tuberculosis antigens between a group of patients with active pulmonary tuberculosis and healthy household contacts
Authors: TORRES M , MENDEZ-SAMPEIRO P , JIMENEZ-ZAMUDIO L , TERAN L , CAMARENA A , QUEZADA R , RAMOS E , SADA E
Published: July 2010

The mycobacterial antigens and the factors related to protection for the development of active tuberculosis are not known. In a natural model of tuberculosis, we studied 10 patients with active pulmonary tuberculosis (non-protective immune response) and 38 healthy household contacts (protective immune response). We tested the lymphocyte proliferative response by T cell Western blotting to eight different antigen fractions and lo two purified mycobacterial antigens of 30 and 64 kD. Patients with active tuberculosis recognized fractions with molecular weights of 80–114, 60–80, 28–41 and 14–19kD. Household contacts recognized the same fractions except the 14–19 kD. The response lo the 64-kD antigen was not significantly different between groups. In contrast, 10% of the patients with active tuberculosis and 73% of the household contacts responded to the 30-kD antigen. The humoral response against the 30-kD antigen by ELISA showed a significantly higher production of antibodies in tuberculosis patients compared with household contacts. We conclude that patients with active pulmonary tuberculosis develop an immune response characterized by poor proliferative response to the 30-kD antigen with a strong humoral response, whereas the opposite occurs in healthy subjects infected by Mycobacterium tuberculoses.
Surface molecules involved in the adherence of recombinant interferon-gamma (rIFN-γ)-stimulated human monocytes to vascular endothelial cells
Authors: WANG J , BEEKHUIZEN H , VAN FURTH R
Published: July 2010

During an inflammatory reaction, an increased number of circulating monocytes adhere to the endothelial cells (EC) of the vessel wall. This process is mediated by molecules located on the surface of monocytes and EC. Locally released Inflammatory mediators can modulate monocyte EC interaction. In an earlier study we reported that stimulation of monolayers of cultured venous EC with rIFN-γ enhanced their adhesiveness for monocytes. The aim of the present study was to investigate the effect of rIFN-γ on peripheral blood monocytes with regard to the expression of surface molecules and the binding to non-stimulated or cytokine-stimulated EC. Flow cytometric analysis demonstrated that monocytes stimulated with 500 U/ml rIFN-γ for 24 h showed increased expression of CR3 (CDIIb;CD18), p150,95 (CD11c/CD18) and Fc-γ (CD64); the expression of LFA-1 (CD11 la/CD18). L-selectin, CD14 and VLA-4 (CD49d/CD29) did not change or was slightly reduced. Upon stimulation with rIFN-γ monocytes showed an enhanced binding to both nonstimulated or rIFN-γ-stimulated EC. This was even more pronounced when EC had been stimulated with rIL-lα for 24 h. The increased binding of rIFN-γstimulated monocytes to rIL-lα-stimulated EC was further analysed. Studies with MoAbs against adhesion molecules on monocytes revealed that the binding of rIFN-γ-stimulated monocytes, hut not that of non-stimulated monocytes, to rIL-lα stimulated EC was inhibited by about 30–60% with MoAbs against CD11a. CDllb, CD18, L-selectin or CD14. MoAbs againsi CD11c or CD49d had little or no effect. From these results, we conclude that exposure of monocytes to rIFN-γ enhances their adhesiveness to cytokine-stimulated EC by a mechanism which involves CD11a/CD18, CD11b/CD18, CD14 and L-selectin on monocytes.
Antibody reactivity to an Epstein–Barr virus BERF4-encoded epitope occurring also in Asp-57 region of HLA-DQ8 β chain
Authors:   THE CHILDHOOD DIABETES IN FINLAND STUDY GROUP, PARKKONEN P , HYÖTY H , ILONEN J , REIJONEN H , YLÄ-HERTTUALA S , LEINIKKI P
Published: July 2010

A five amino acids-long sequence (GPPAA) in the region of the 57th amino acid of HIA-DQ8 β chain, with seems to be important in defining the risk for type I diabetes, occurs also in the BERF4-encoded EBNA 3C protein of Epstein-Barr virus (EBV) in six successive repeats. The antigenicity of this region was analysed using synthetic peptides containing different modifications of the GPPAA sequence. Two of the seven individuals who had acute EBV infection produced antibodies against an EBV-derived peptide (GPPAAGPPAAGPPAA) paralleling the EBNA2 antibodies. These two cases also contracted type I diabetes immediately after the infection. High antibody levels against this peptide were found in a total of 12%, of EBV+ individuals, and in most cases antibodies remained at high levels for several years. Human sera as well as affinity-purified antibodies specific for the GPPAAGPPAAGPPAA peptide reacted also with shorter peptide analogues (GPPAAGPPAA and GPPAA), as well as with peptides containing the surrounding motifs from DQ8 β chains. However, none of these antibodies bound to denatured DQ8 β chains in immunoblotting. The charge of the 57th amino acid modulated the antigenicity of this epitope, as peptides from Asp-57-negative DQ molecules were reactive, while peptides from Asp-57-positive DQ molecules were not. The responsiveness was seen in both HLA-DQ8-positive and -negative subjects as well as in type I diabetic individuals. The results suggest that some individuals who carry the GPPAA sequence in their HLA-DQ molecule recognize this epitope in EBV. This phenomenon may have potential importance in EBV-induced immune abnormalities, although cross-reactivity against DQ molecules could not be demonstrated in the present study.
IgG2 subclass restriction of anti-β2 glycoprotein 1 antibodies in autoimmune patients
Authors: ARVIEUX J , ROUSSEL B , PONARD D , COLOMB M G
Published: July 2010

The IgG subclass and light chain distribution of antiphospholipid antibodies (aPL) occurring in autoimmune patients were determined by means of two radioimmunoassays using either cardiolipinor β2 glycoprotein 1 (β2GP1)-coated microtitre plates and mouse MoAbs. Of 50 sera selected for positivity of anticardiolipin antibodies (ACA) of the IgG isotype, 32 (64%) possessed anti-β2GPl antibodies and their presence was closely associated with clinical features of the antiphospholipid syndrome. Good correlations were found between ACA and anti-β2GP1 antibodies when considering antibody level and patterns of light chain and IgG subclass, suggesting that, overall, the same antibodies were being measured. Light chain analysis showed the polyclonal origin of these antibodies and, in most sera, a trend towards use of λ chain. Among sera positive for anti-β2GP1 antibodies, IgG2 was the major subclass reactive with β2GP1 and cardiolipin (87% and 74β2 of the IgG antibody activity, respectively). In contrast, in the group of 18 sera lacking anti-β2GP1 antibodies, ACA were largely restricted to lgG3, with a lesser contribution by IgGl. A few selected sera from the anti-β2GP1-positive group were shown to contain mixtures of antibodies that required β2GP1 (restricted to IgG2 present in large amounts) and did not require this cofactor (restricted to IgG3 and/or IgG1 present in low amounts) for their reactivity with cardiolipin. There was no contribution of glycosylation to the epitopes recognized by anti-β2GP1 antibodies, even though human anti-carbohydrate antibodies are restricted to the IgG2 subclass. These findings further emphasize the intra- and interindividual heterogeneity of aPL, and should help to discriminate clinically relevant specificies.