bluuebunny/crossref_metadata_2025 huggingface dataset:
The level of the serum opsonin, mannan-binding protein in HIV-1 antibody-positive patients
Authors: NIELSEN S L , ANDERSEN P L , KOCH C , JENSENIUS J C , THIEL S
Published: July 2010

The concentrations of mannan-binding protein (MBP) in consecutive samples from 10 HIV+ persons were estimated using an ELISA based on polyclonal rabbit anti-MBP. The changes in MBP with time were similar in HIV+ and HIV− persons, and did not appear to be of clinical significance. MBP was determined in a further 70 persons found HIV-1+ during a period af 2·5 years (1984–1986). Out of the total of 80 patients, 32 have by now died from AIDS. According to the serum level of MBP the HIV-infected persons were grouped into high (> 650 ng MBP/ml). intermediate (101–650 ng/ml), and low MBP (< 101 ng/ml). At the termination of the study the frequency of deaths/total in each of the groups were: high MBP, 14/39 (36%); intermediate MBP, 12/26 (46%); and low MBP, 6/14 (43%). There was no association between the MBP level of the individual and the progressive loss of CD4+ T cells, and the level of MBP was not predictive for the length of time between the detection of HIV antibodies and development of AIDS, nor for the duration of AIDS before death occurred. The number of HIV+ persons without detectable MBP (10%) was significantly higher than previously reported for healthy persons (2·4%, P= 0·027). The course of HIV infection does not seem to be influenced by the level of MBP, nor does the antimicrobial activity of MBP appear to affect the progression of AIDS. Further studies are required to substantiate the significance of absence of MBP in the susceptibility to HIV.
Inhibition ofex vivoproinflammatory cytokine secretion in fatalMycobacterium tuberculosisinfection
Authors: FRIEDLAND J S , HARTLEY J C , HARTLEY C G C , SHATTOCK R J , GRIFFIN G E
Published: July 2010

Tuberculosis is characterized by fever, weight loss, a prolonged acute-phase protein response and granuloma formation. These characteristics may partly be due to action of proinflammatory cytokines tumour necrosis factor (TNF), IL-6 and IL-8. We investigated plasma concentrations of these cytokines before and after ex vivo lipopolysaccharide stimulation of whole blood leucocytes from 41 Zambian patients with tuberculosis, 32 of whom were also HIV+. Although patients had a reduced weight, were more anaemic and had higher erythrocyte sedimentation rate compared with controls (all P < 0·0005), clinical and laboratory measurements of disease state were similar in those who died and survivors. In contrast, plasma IL-6 and IL-8 concentrations were higher in patients who died (P < 0·05). There was no detectable cytokine mRNA in unstimulated leucocytes. There was reduced secretion of TNF (P < 0·005 at 2h), IL-6 (P < 0·005 at 8 h) and IL-8 (P < 0·005 at 24 h) after ex vivo stimulation of whole blood leucocytes from patients who died compared with survivors. This was partly due to a soluble inhibitory factor present in plasma. The only additional effect of concurrent infection by HIV with Myco. tuberculosis was decreased IL-6 secretion following ex vivo stimulation of leucocytes. Reduced proinflammatory cytokine release may represent a critical impairment of host immune defences important in determining outcome in tuberculosis.
Synthesis and regulation of accessory/proinflammatory cytokines by intestinal epithelial cells
Authors: PANJA A , SIDEN E , MAYER L
Published: July 2010

Intestinal epithelial cells (IEC) have been shown to act as antigen-presenting cells (APC) in vitro and may have this capacity in vivo. In order to determine whether IEC, like other APC, are able to produce accessory cytokines which may play a role in T cell activation, we assessed the accessory cytokine profile of IEC constitutively or after stimulation. We measured expression, production and regulation of accessory cytokines (IL-1β, IL-6, tumour necrosis factor-alpha (TNF-α), transforming growth factor-beta (TGF-β) by the presence of mRNA as well as secreted protein. Freshly isolated IEC from surgical specimens were cultured in the presence or absence of lipopolysaccharide (LPS), interferon-gamma (IFN-γ), IL-1β or TNF-α. mRNA was assessed by a specific RNAse protection assay which controlled for contaminating cell populations while protein secretion was measured by ELISA (IL-1) or bioassay (TNF and IL-6). Neither IL-1β nor TNF-α were detectable in cultured IEC supernatants, supporting the lack of macrophage contamination. All IEC spontaneously secreted IL-6 at levels comparable to those of macro-phages. IEC IL-6 mRNA also increased approximately 200-fold during the first 24 h of culture. LPS, IFN-γ or TNF-α had no effect on spontaneous IL-6 production, and neither resulted in the secretion of IL-1β or TNF-α. However, IL-1β up-regulated IL-6 synthesis by 6–7-fold. IEC express a profile of cytokine mRNAs distinct from conventional APC (low level constitutive IL-6 expression but no detectable IL-1β, TGF-β or TNF-α), adding to their uniqueness as APC.
IL-1 receptor antagonist (IL-1Ra) does not inhibit the production of C-reactive protein or serum amyloid A protein by human primary hepatocytes. Differential regulation in normal and tumour cells
Authors: GABAY C , GENIN B , MENTHA G , IYNEDJIAN P B , ROUX-LOMBARD P , GUERNE P-A
Published: July 2010

The synthesis of some class 1 acute-phase proteins (APP), including C-reactive protein (CRP) and serum amyloid A (SAA) protein is completely blocked by the IL-1 receptor antagonist (IL-1Ra), whereas the production of fibrinogen, a class 2 APP, is increased by IL-1Ra in hepatoma cells, but this has never been tested in human hepatocytes in primary culture. Since previous studies on the contributions of cylokine inhibitors in connective tissues diseases suggested that IL-1 and tumour necrosis factor-alpha (TNF-α) might play an important role in the regulation of CRP, we decided to examine in more detail the respective roles of IL-1β, IL-6, and TNF-α and their inhibitors in the production of APP by human primary hepatocytes versus the hepatoma cell line PLC/PRF/5. In the hepatoma cell line, IL-1β and/or TNF-α had synergistic effects with IL-6 on the production of CRP and SAA. In contrast, these cytokines were devoid of effect in normal hepatocytes. The production of fibrinogen was increased by IL-6 and decreased by IL-1 (and TNF-α) in both cell types. The secretion of CRP and SAA by primary hepatocytes incubated with a cytokine-rich mononuclear cell-conditioned medium was totally unaffected by IL-1Ra or anti-TNF-α antibodies. In contrast, the addition of IL-1Ra increased the production of fibrinogen by both hepatoma cells and primary hepatocytes incubated with the mononuclear cell-conditioned medium. We therefore conclude that IL-1β and TNF-α do not exert any significant effect on the synthesis of CRP and SAA by human primary hepatocytes.
Immune responsiveness to inhaled antigens: local antibody production in the respiratory tract in health and lung diseases
Authors: YOSHIZAWA Y , OHTANI Y , INOUE T , MIYAKE S , IKEDA A , TANBA M , KURUP V P , MIYAMOTO K
Published: July 2010

The pulmonary defence mechanism in summer type hypersensitivity pneumonitis induced by Trichosporon cutaneum was investigated. We have studied the antibody response to fungal antigens in sera and bronchoalveolar lavage (BAL) fluids from patients with summer type hypersensitivity pneumonitis, bird fancier's lung, interstitial pneumonia associated with collagen vascular disease (INT-PNE), and from normal volunteers. Antigens extracted from fungi frequently isolated from home environments were used in ELISA to detect IgG and IgA antibodies in sera and BAL fluids. The results of the present study show that antibody titre in the respiratory tract to a variety of fungi from home environments is modulated by ongoing pulmonary inflammation, and that antibody production against inhaled antigens is altered by pulmonary inflammation resulting from diverse pathogenesis. This study concludes that the preexisting pulmonary inflammatory disease alters antibody production in the respiratory tract in response to inhaled fungi, and that the type of alteration depends in part on the etiology of the preexisting disease.
The inhibitory effects ofMycobacterium tuberculosison MHC class II expression by monocytes activated with riminophenazines and phagocyte stimulants
Authors: WADEE A A , KUSCHKE R H , DOOMS T G
Published: July 2010

The expression of MHC class II antigens by peripheral blood monocytes from normal individuals was investigated. Class II expression as determined by a cell HLISA was effectively induced by various phagocyte stimulants. A further aspect of our study investigated the effects of clofazimine, a riminophenazine antimicrobial agent and its analogue, B669, on class II expression. Both agents at concentrations attainable in vivo increased the expression of MHC class II antigens. A 25-kD glycolipoprotein derived from Mycobacterium tuberculosis that inhibits phagocyte functions has previously been described. This component significantly reduced the expression of MHC class II antigens induced by the riminophenazines, clofazimine and B669, interferon-gamma (IFN-γ) or opsonised yeast when added at the initiation of experiments. The riminophenazines could not restore the decrease in class II antigen expression previously inhibited by the 25-kD mycobacterial fraction. However, cultures prestimulated with the riminophenazines or phagocyte stimulants were unaffected by the 25-kD mycobacterial fraction. The results suggest the potential use of these agents as modulators of phagocyte function.
Ketotifen therapy in chronic graft-versus-host disease (cGVHD): effect on mast cells and fibroblasts
Authors: NAGLER A , SEGAL V , SLAVIN S , LEVI-SCHAFFER F
Published: July 2010

Current treatment options for cGVHD are limited. Mast cells (MC) and fibroblasts have been shown to play a role in the murine model of cGVHD. Ketotifen is an anti-H-1 antihistamine with MC-stabilizing properties. We therefore treated eight patients with cGVHD with ketotifen (6μg/ day for 3 months). Three additional age- and sex-matched cGVHD patients served as controls. MC count and activation state in cGVHD skin biopsies and the in vitro effect of peripheral blood mononuclear cell (PBMC) supernatants on (i) histamine release by MC; (ii) 3T3-fibroblast proliferation; and (iii) prostaglandin E2 (PGE2) production, were evaluated. Ketotifen therapy resulted in clinical improvement in 4/8 patients, stabilization of the disease in 2/8, while in 2/8 patients the cGVHD progressed and they died of bacterial sepsis. Side effects were minimal. In the skin biopsies the number of MC was found to be 0.58±0.17 (n= 8) (field ± 400) and the MC looked degranulated (toluidine blue staining). Following ketotifen therapy MC number was increased to 1.2±0.28 (n= 8) (P < 0.05). PBMC supernatants of cGVHD patients stimulated histamine release from cultured rat MC(n= 8)(2.7±0.5 μg/ml; normal values are 2±1.0±4 μg/ml, n = 5). Ketotifen therapy reduced the histamine release level to the normal range (2.0±0.5 μg/ml, P < 0.05) (n= 8). Ketotifen therapy had no significant effect on; (i) 3T3 fibroblast proliferation which was suppressed by cGVHD PBMC supernatants; (ii) the elevated PGE2 production which we observed when fibroblasts were incubated with the PBMC supernatants. These results indicate that ketotifen may play a therapeutic role in cGVHD.
Dengue virus-induced human cytotoxic factor: production by peripheral blood leucocytesin vitro
Authors: Mukerjee R , Chaturvedi U C , Dhawan R
Published: July 2010

During dengue type 2 virus (DV) infection of mice a unique cytokine, the cytotoxic factor (CF), is produced which reproduces the pathological lesions seen in patients of dengue haemorrhagic fever (DHF). Recently we have observed a CF-like protein in the sera of DHF cases. The present study was undertaken to investigate whether DV can stimulate human peripheral blood mononuclear cells (PBMC) in vitro to produce human CF (hCF). Cultures prepared from PBMC or its enriched subpopulations were inoculated with 1000 LD50 of DV and controls with normal mouse brain suspension (NMB). Aliquots of cultures were harvested daily from 24 h to 96 h and their supernatant (CS) and cells were separated. CS were screened for viral titres and for the presence of hCF by cytotoxicity assay, inhibition ELISA, dot blot and Western blot tests using anti-mouse-CF antibodies. The RNA from the cells was screened in Northern blot and dot blot tests for the presence of mRNA for CF. It was observed that hCF appeared in the CS of DV-infected culture of PBMC and T-enriched cells at 48 h and was present until 96 h; no CF was detected in CS of B cells or monocyte cultures. The production of hCF was abrogated by pretreatment of the T cells with anti-CD4 antibodies but not with anti-CDS antibodies, indicating that hCF was produced by CD4 T cells. The Northern blot analysis using oligonucleotide probes prepared on the basis of amino-terminal sequence of mouse CF, showed presence of mRNA for hCF in PBMC and T cell cultures. DV replicated in PBMC cultures with peak titres at 48 h. The findings of the present study demonstrate that DV-induced hCF is produced by human T cells.
Insulin-like growth factor-1 (IGF-1) protects NOD mice from insulitis and diabetes
Authors: Bergerot I , Fabien N , Maguer V , Thivolet C
Published: July 2010

To evaluate the effect of IGF-1 on the autoimmune process of d cell destruction, permissive non-obese diabetic (NOD) recipients were adoptively transferred with 7 × 106 autoreactive T cells from diabetic NOD mice and were administered subcutaneously 10μ;g rhIGF-1, twice daily for 3 weeks. Administration of rhIGF-1 reduced the final incidence of successful transfers of diabetes observed in only 6/24 mice (25%) versus 12/21 (57%) in control mice. A marked reduction of insulitis during histological analysis of pancreatic glands was also observed. Mice treated with rhIGF-1 had a higher percentage of intact islets (486 ± 12% versus 16 ± 1·1%. P= 0·001) and a lower percentage of infiltrated islets. Islets from rhIGF-1-treated mice had a more intense insulin staining reflecting a higher β cell mass, but no difference was observed in the amount of insulin content of pancreatic extracts and in the amounts of mRNA transcripts for proinsulin. No difference was also observed in the titres of three islet cell antibody (ICA)-positive sera and in the pattern of A2B5 staining. Some mice developed diabetes and severe islet cell infiltration despite rhIGF-1, thus indicating that some committed T cells were still able to invade the islets and cause β cell destruction. The percentages of CD4+ and CD8+ T cells in the spleen of experimental mice were similar. To evaluate the effects of rhIGF-1 on cell trafficking in recipient mice, T cells from diabetic NOD Thy-1,2 mice injected into congenic NOD-N Thy-1,1 mice were monitored 3 weeks after adoptive cell transfer. The percentage of Thy-1,2+ T cells was significantly reduced in the spleen (10·8 ± 1·3%versus 17·2 ± 3·9%, P= 0·004) of rhIGF-1 treated mice in contrast to the thymus (68·4 ± 7·9%versus 72·87 ± 6·2%, P= 0·306), suggesting that rhIGF-1 could influence T cell trafficking to the lymphoid organs. The findings that rhIGF-1 has protective effects in autoimmune diabetes opens new perspectives for future experiments as well as for preventive strategies in human type I diabetes.
Allelic polymorphism in IL-1β and IL-1 receptor antagonist (IL-1Ra) genes in inflammatory bowel disease
Authors: Bioque G , Crusius J B A , Koutroubakis I , Bouma G , Kostense P J , Meuwissen S G M , PeÑA AS
Published: July 2010

Recent reports have shown that allele 2 of the IL-1 receptor antagonist (IL-1Ra) gene is over-represented in ulcerative colitis (UC). Healthy individuals carrying allele 2 of this gene have increased production of IL-1Ra protein. Since the final outcome of the biological effects of IL-1β may depend on the relative proportion of these two cytokines, we have studied if a TaqI polymorphism in the IL-1β gene, which is relevant to IL-1β protein production, may be involved in the genetic susceptibility to UC and Crohn's disease (CD), in association with the established IL-1Ra gene polymorphism. Polymorphisms in the closely linked genes for IL-1β and IL-1Ra were typed in 100 unrelated Dutch patients with UC, 79 with CD, and 71 healthy controls. The polymorphic regions in exon 5 of the IL-1β gene and in intron 2 of the IL-1Ra gene, were studied by polymerase chain reaction (PCR)-based methods. The IL-1β allele frequencies in UC and CD patients did not differ from those in healthy controls. In order to study if the IL-1β gene polymorphism might participate synergistically with the IL-1Ra gene polymorphism in susceptibility to UC and CD, individuals were distributed into carriers and non-carriers of allele 2 of the genes encoding IL-1β and IL-1Ra, in each of the patient groups and controls. Results indicated a significant association of this pair of genes, estimated by the odds ratio (OR) after performing Fisher's exact test, in the UC group (P= 0·023, OR = 2·81), as well as in the CD group (P= 0·01, OR = 3·79). Thus, non-carriers of IL-1β allele 2 were more often present in the subgroup of patients carrying the IL-1Ra allele 2. By contrast, no association of these alleles was detected in the group of healthy controls (P= 1·00, OR = 0·92). These results suggest that the IL-1β/IL-1Ra allelic cluster may participate in defining the biological basis of predisposition to chronic inflammatory bowel diseases.
Dual action of IL-4 on mite antigen-induced IgE synthesis in lymphocytes from individuals with bronchial asthma
Authors: Kawano Y , Noma T
Published: July 2010

Polyclonal IgE synthesis was efficiently induced by Dermatophagoides farinae (Df) antigen in freshly derived peripheral blood lymphocytes from mite-sensitive individuals with bronchial asthma. The in vitro IgE production was significantly correlated with total serum IgE values. The induced IgE synthesis was inhibited in a dose-dependent manner by antibodies to IL-4, indicating a role for endogenous IL-4. Although IL-4 alone increased IgE production, high concentrations (1–100 U/ml) of the cytokine inhibited Df antigen-stimulated production of IgE. However, low concentrations (0·001-0·01 U/ml) of IL-4 significantly enhanced Df antigen-induced IgE production, as did high doses of IL-4 when endogenous IL-4 was neutralized by antibodies to IL-4. Two other cytokines, IL-10 and interferon-gamma (IFN-γ), showed contrasting actions, as judged from experiments with exogenous cytokine and anti-cytokine antibodies: IL-10 enhanced and IFN-γ inhibited Df antigen-induced IgE synthesis. Thus, mite-stimulated IgE production in lymphocytes from individuals with bronchial asthma appears to be regulated by at least three cytokines; TL-4, IL-10, and IFN-γ.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) reduces toxoplasmastatic activity of human monocytes via induction of prostaglandin E2 (PGE2)
Authors: Delemarre F G A , Stevenhagen A , Van Furth R
Published: July 2010

In this study we investigated the effect of human GM-CSF on the toxoplasmastatic activity and release of H2O2 and PGE2 by human monocytes. Incubation of monocytes from healthy controls with GM-CSF resulted in a dose-dependent reduction of toxoplasmastatic activity and a decrease in H2O2 production. Furthermore GM-CSF-treated monocytes released more PGE2 than untreated cells. To investigate the role of PGE2 in the reduced toxoplasmastatic activity of GM-CSF-treated monocytes, these cells were incubated with indomethacin. This resulted in a reduction of PGE2 release and restoration of toxoplasmastatic activity of monocytes treated with GM-CSF. GM-CSF reduces the toxoplasmastatic activity of monocytes via production of PGE2.
Lipopolysaccharide (LPS) down-regulates CD4 expression in primary human macrophages through induction of endogenous tumour necrosis factor (TNF) and IL-1β
Authors: Herbein G , Doyle A G , Montaner L J , Gordon S
Published: July 2010

The regulation of CD4 expression on macrophages and its role in immune cell interactions remain obscure. In contrast with primary lymphocytes, primary macrophages express only low amounts of surface CD4, which is regulated differentially for example by adherence in vitro. We report that addition of LPS for 1–5 days to human blood monocyte tissue culture-derived macrophages (TCDM) down-regulates both surface CD4 expression and total cellular CD4 antigen content as measured by flow cytometry and Western blot analysis. TNF-α and IL-1β, proinflammatory cytokines which are both induced by LPS. also down-regulate surface and total CD4 expression in TCDM, This down-regulation of CD4 expression by LPS, TNF-α, and IL-1β occurs at the level of transcription. The decreased macrophage CD4 expression induced by LPS was blocked by MoAbs directed against human TNF-α and IL-1β, demonstrating that LPS acts on CD4 expression through induction of endogenous TNF-α and IL-1β. Conversely, neither LPS nor TNF-α and IL-1β were able to modulate surface CD4 expression on quiescent or phytohaemagglutinin (PHA)-activated lymphocytes. Of other cytokines and growth factors tested, Th2 cytokines (IL-4. IL-10, IL-13), chemokines (MCP-1, MIP-1α, RANTES), and macrophage colony-stimulating factor did not alter CD4 expression in primary macrophages, granulocyte-monocyte colony-stimulating factor and the prototypal Th1 cytokine interferon-gamma (IFN-γ) modulated surface CD4 expression only after prolonged treatment (5 days). Our results show that LPS. TNF-α and IL-1β selectively down-regulate CD4 expression in primary human macrophages, and that decreased CD4 expression induced by LPS results from endogenous secretion of TNF-α and IL-1β by the macrophages.
T cell activation byTheileria annulata-infected macrophages correlates with cytokine production
Authors: Brown D J , Campbell J D M , Russell G C , Hopkins J , Glass E J
Published: July 2010

A major feature of the pathology induced by Theileria annulata is acute lymphocytic proliferation, and this study investigates the mechanisms underlying the intrinsic ability of T. annulata-infected monocytes to induce naive autologous T cells to proliferate. Different T. annulata-infected clones expressed different but constant levels of MHC class II, varying from < 1.0 × 105 to 1.5 × 106 molecules/cell, as measured by saturation binding. However, no correlation was found between the level of MHC class II expression and levels of induced T cell proliferation. Theileria anmulata infected cell lines and clones were assayed for cytokine mRNA expression by reverse transcriptionpolymerase chain reaction (RT-PCR). The infected cells assayed produced mRNA specific forIL-lα, IL-1β, lL-6, IL-10 and tumour necrosis factor-alpha (TNF-a). but not IL-2 or lL-4. One clone (clone G)did not produce mRNA for TNF-α. The degree of T cell proliferation induced by infected cells was directly correlated with the amount of mRNA produced for the T cell stimulatory cytokines IL-lα and IL-6, as assessed by a semiquantitative technique. In contrast, cells infected with the related parasite T. parva produced mRNA for IL-lα, IL-2, IL-4, IL-10 and interferon-gamma (IFN-γ). Since T. parva-infected cells also induce naive autologous T cell proliferation, it seems likely that the production of lL-1α by cells infected with either parasite is a major signal for the induction of non-specific T cell proliferation.
Phenotypic characterization of two cell populations involved in the acquisition of suppressor activity by cultured spleen cells fromMycobacterium lepraemurium-infected mice
Authors: Gosselin D , Turcotte R , Lemieux S
Published: July 2010

The impairment of cellular immunity in mice infected with Mycobacterium lepraemurium was shown to correlate with the development of suppressor cells. We have previously reported that before suppressor activity is detectable in freshly harvested cell suspensions, suppressor cell precursors accumulate in the spleen of infected mice. Upon overnight culture in the presence of a regulatory cell subset, these precursor cells acquire the capacity to impair the concanavalin A (Con A)-induced proliferation of normal spleen cells. The purpose of this study was to determine the phenotype of the cells involved in this phenomenon. This was done by following the development of suppressor activity in spleen cell suspensions depleted of defined cell subsets of the adherent or the non-adherent cell fractions with selected MoAbs and immunomagnetic beads or by in vivo treatment. Our results indicate that the acquisition of suppressor activity requires the interaction of Ia+CD11b+FcγR+IgG- asialo GMI- adherent cells with Thyl 1-CD4-CD8-lgG- Ia- asialo GMl -FcγR+CD1 lb+ non-adherent cells. It is also shown that the development of suppressor activity is impaired by preventing cell-cell contact between these two cell subsets through coculture in ‘Transwell chambers’. These observations support the conclusion that the in vitro acquisition of suppressor activity is a consequence of the maturation of suppressor cell precursors of the monocytic lineage induced by a receptor-ligand type interaction with a non-adherent cell subset that is clearly distinct from mature T, B and natural killer (NK) cells.
Serum soluble markers in the evaluation of treatment in human visceral leishmaniasis
Authors: Schriefer A , Barral A , Carvalho E M , Barral-Netto M
Published: July 2010

Visceral leishmaniasis (VL) has a fatal course if not properly treated. Recovery from VL is linked to cellular immune response. Unresponsiveness to antimonial therapy reinforces the importance of determining parameters for treatment assessment. We analysed the pre- and post-treatment serum levels of solubleCD4(sCD4).sCD8. SI L-2R, soluble intercellular adhesion molecule-1 (sICAM-1) and neopterin in groups of VL patients either responsive or not to standard antimonial therapy. Pretreatment serum levels of all markers except for sICAM-1 were significantly higher in VL patients than in healthy subjects from the same area (P < 0 05). sICAM-1 levels were similar in healthy controls and in VL patients refractory to antimonial therapy (p = 0 25), but significantly higher in patients responsive to treatment (p = 0.02). The comparison of pre- and post-treatment concentrations showed that all markers, except sCD4 and sICAM-1, presented a significant fall (p < 0. 05) in patients responsive to antimonial therapy. However, only neopterin presented with levels compatible with those of healthy subjects at the end of treatment (p = 0. 30). In refractory patients sICAM-1 presented with post-treatment levels significantly higher than the pretreatment determinations (p = 0.03). while sCD4 experienced a significant drop (p = 0. 01) All markers displayed clearly distinct behaviour according to the patient's response to therapy. This makes all soluble molecules studied suitable for use as indicators of antimonial therapy response. Additionally the comparison of pretreatment levels of the markers between responders and refractory patients to antimonial therapy showed that serum concentrations of sIL-2R and sICAM-1 significantly differed among these two groups (p=0.02 in each case), suggesting that they may be used in future as predictors of antimonial therapy response.
Changes in the phenotype of monocytes/macrophages and expression of cytokine mRNA in peripheral blood and synovial fluid of patients with rheumatoid arthritis
Authors: Highton J , Carlisle B , Palmer D G
Published: July 2010

Data from a previous study suggested that peripheral blood monocytes in patients with rheumatoid arthritis (RA) may be activated. Therefore, in this study we sought further evidence of ‘presynovial’ activation of monocytes. Our results show that phenotypic changes are demonstrable in peripheral blood monocytes in patients with RA, including increased expression of CR3 (CDllb/CD18) and FcRI (CD64). However, changes are most extensive in synovial monocytes/macrophages and especially for HLA-DR and intercellular adhesion molecule-1 (ICAM-1) (CD54). We conclude that monocyte/macrophage activation is most evident within the joint, and that ‘presynovial’ changes occur but are of limited extent.
Differential effects of methotrexate and liposomally conjugated methotrexate in rat adjuvant-induced arthritis
Authors: Williams A S , Camilleri J P , Amos N , Williams B D
Published: July 2010

In this study we evaluated the comparative efficacy of free and liposomally conjugated methotrexate on both disease indtiction and suppression of acute inflammation in rat adjuvant-induced arthritis. Rats were given either empty liposomes (E-LIPO), free methotrexate (MTX) or the liposomally conjugated methotrexate (MTX-LIPO) at a dose of 100/μg/day for 7 consecutive days by the intravenous route. When MTX treatment was initiated on the day of arthritis induction the drug suppressed bul did not abolish the development of joint inflammation. Free MTX had no significant anti-inliammatory effect upon an established arthritis when dosing was commenced on day 11 post-adjuvant induction. Conversely, MTX-LIPO did not affect the progression of the arthritis when dosing was started on day 0, but exerted a significant anti-inflammatory effect on an established arthritis. MTX-LIPO treatment was significantly less haematotoxic than free MTX.
Comparison of activation marker and TCR Vβ gene product expression by CD4+ and CD8+ T cells in peripheral blood and lymph nodes from HIV-infected patients
Authors: Ramzaoui S , Jouen-Beades F , Michot F , Borsa-Lebas F , Humbert G , Tron F
Published: July 2010

Since lymphoid organs constitute the site of active and progressive HIV disease, analysis of their lymphocytes may provide more accurate information on T cell abnormalities than that obtained from studying peripheral blood lymphocytes. The objective of this study was to compare the expressions of activation markers and T cell receptor (TCR) Vβ gene products by CD4+ and CD8+ T cells in lymph nodes (LN) and peripheral blood (PB) from healthy individuals and asymptomatic HIV-infected patients to determine whether anomalies that could be identified at the HIV replication site could support the hypothesis of T cell activation by HIV-encoded antigens or superantigens. CD4+ and CD8+ T cells in paired LN and PB obtained from six healthy controls and five asymptomatic HIV-infected individuals were analysed by flow cytometry, using anti-CD38, anti-HLA-DR and 13 anti-Vβ MoAbs that cover, approximately, 45% of the T cell repertoire. Analysis of T cell activation marker expression indicated that the percentages of CD4+ and CD8+ T cells bearing CD38 or CD38 and HLA-DR molecules were higher in patients than in controls and, in patients, higher in LN than in PB. Comparison between the Vβ repertoires of CD4+ and CD8+ T cells in LN and PB showed that, in each healthy individual, a limited number of Vβ families expressed by CD4+ or CDS+ T cells had different repartition in LN and PB, whereas in each HIV+ patient, more Vβ families exhibited different distributions and these differences recurred among certain Vβ segments, such as Vβ5.3 and Vβ21 in the CD4+ T cell population and Vβ5.2/5.3, Vβ12 and Vβ12 in the CD8+ T cell population. Taken together, these data argue for a skewed TCR repertoire in HIV infection and sustained activation of T cells by HIV-encoded antigens at the site of HIV replication, and further demonstrate that a high proportion of CD4+ T cells are in an activation state that may, indirectly, participate in their functional abnormalities.
Theileria annulatainduces abberrant T cell activationin vitroandin vivo
Authors: Campbell J D M , Howie S E M , Odling K A , Glass E J
Published: July 2010

The protozoan parasite of cattle, Theileria annulata, causes a severe lymphoproliferative disease, developing initially in the draining lymph node, which is often fatal in naive animals. Infection of macrophages with T. annulata leads to an augmentation of their antigen-presenting capability in vitro and infected cells can induce proliferation of autologous resting T cells from naive animals. This inappropriate activation of T cells may play an important role in the failure of the host to mount an effective immune response in vivo. To investigate this hypothesis we characterized further the response of T cells from naive cattle to infected cells in vitro, and also examined the development of the immune response in lymph nodes draining the sites of T. annulata infection. Both CD4+ and CD8+ T cells from naive peripheral blood mononuclear cells (PBMC) were induced to proliferate and express the activation markers IL-2R and MHC class II when cultured with infected cells. This effect was seen in both ‘naive’ and ‘memory’ T cells, and was dependent upon contact with infected cells. In vitro, infected cells are therefore capable of activating T cells irrespective of their antigen specificity or memory status. In draining lymph nodes, although large numbers of IL-2R+ cells developed following infection, these activated cells were only associated with areas of parasite-induced proliferating cells, and subsequently disappeared from the node. Cells expressing IL-2R were not present in recognized sites for T cell development. Germinal centres were severely affected, losing T cell-dependent zones followed by a total destruction of morphology. T cell function is therefore severely disrupted within draining nodes. This study has shown that parasitized cells supply sufficient signals in vitro to activate T cells irrespective of specificity. T cells also are not stimulated in a conventional manner in vivo, and this may play an important role in preventing an effective immune response from being generated.
Characterization of CD4+ T helper cells in patients with Kawasaki disease (KD): preferential production of tumour necrosis factor-alpha (TNF-α) by Vβ2− or Vβ8− CD4+ T helper cells
Authors: Sakaguchi M , Kato H , Nishiyori A , Sagawa K , Itoh K
Published: July 2010

KD is an acute febrile illness in children characterized by coronary arteritis accompanied by aneurysm and thrombotic occlusion. The etiology of KD is unknown. It has been recently reported that KD is associated with the selective expansion of Vβ2+ and Vβ8.1+ T cells in peripheral blood lymphocytes (PBL), by studying the T cell receptor (TCR) repertoire of in vitro activated T cells. KD may therefore be caused by a superantigen [1–3]. To understand better the immunopathology of KD, we investigated TCR Vβ2 and Vβ8.1 expression on both the T cells of freshly isolated PBL and T cell clones (TCC) from patients with KD. Cytokine production by TCC was also studied. Blood samples were obtained from patients with acute (n= 20) and convalescent (n= 20) KD, agematched children with non-infectious diseases (n= 18), and healthy adults (n= 20). Among these four groups, there were no significant differences in the percentages of either Vβ2+ or Vβ8.1+. T cells of freshly isolated PBL. The same was true for the CD4+ or CD8+ T cell subsets. One hundred and five TCC (98 CD3+ CD4+ CD8− and seven CD3+ CDA− CD8+) established from the affected skin, lymph node or PBL of six patients with KD were also negative for either Vβ2 or Vβ8.1 TCR. Sixty-eight of 105 TCC (65%) produced detectable levels (>5 pg/ml) of TNF-α (6–1016 pg/ml), in the absence of any stimuli. In contrast, only 11 (10%) of 105 TCC or 7(7%) of 97 TCC produced detectable levels of IL-2 or IL-6, respectively, in the absence of any stimuli. Stimulation with phytohaemagglutinin (PHA) and phorbol myristate acetate (PMA) induced most TCC to produce higher amounts of TNF-α, IL-2 and IL-6. These results suggest that CD4+ T helper cells expressing TCR-β other than Vβ2 or Vβ8 receptor, primarily through TNF-α production, are involved in the immunopathology of KD.
Passive dual immunization against tumour necrosis factor-alpha (TNF-α) and IL-1β maximally ameliorates acute aminonucleoside nephrosis
Authors: MartÍN A , Molina A , Bricio T , Mampaso F
Published: July 2010

Rats receiving a single dose (10mg/100g) of aminonucleoside of puromycin (PAN) develop heavy proteinuria and acute interstitial nephritis (AIN). Whole isolated glomeruli from rats injected with PAN secreted both TNF-α and IL-1β cytokines. TNF-α secretion was first and maximally detected on day 3, whereas IL-β activity was found on day 7, when rats were heavily proteinuric and AIN developed. In vivo treatment with either anti-TNF-α or anti-IL-1β antibodies produced a drastic and simultaneous reduction in both levels of proteinuria and intensity of interstitial cell infiltrate. These effects improved when both antibodies were administered together. Our studies demonstrate the effectiveness of immunosuppressive therapy against these two cytokines in rats with PAN-induced nephrosis.
Relationship between facilitated allergen presentation and the presence of allergen-specific IgE in serum of atopic patients
Authors: Van Der Heijden F L , Van Neerven R J J , Kapsenberg M L
Published: July 2010

Allergen presentation to allergen-specific T cells can be facilitated when IgE–allergen complexes are endocytosed by antigen-presenting cells (APC) after binding to the low-affinity Fc·R type II (CD23). Here we present a study on the relative capabilities of sera of atopic patients to mediate facilitated antigen presentation (FAP). To this aim FAP was studied in an in vitro model in which CD23-expressing Epstein–Barr virus (EBV)-B cells act as APC to T lymphocyte clones (TLC) that are specific for Der p 2, a major allergen of housedust mite Dermatophagoides pteronyssinus (Dp). Der p 2 is immune-complexed by preincubation in sera from atopic patients, containing allergen-specific IgE. If EBV-B cells are preincubated with these complexes before using the cells as APC, the allergen-specific TLC proliferate at 100–1000-fold lower allergen concentration than required for T cell activation after presentation of uncomplexed allergen. The relative capability of various sera to mediate FAP was correlated with total serum IgE, and especially with Der p 2-specific serum IgE. In the model used, a high FAP capacity could be demonstrated only in sera with a total serum IgE concentration above approximately 2 /μg/ml or with Der p 2-specific IgE above approximately 100 ng/ml. Maximal FAP, i.e. the ability to induce maximal proliferation of the TLC, was obtained in the presence of more than × 600 ng Der p 2-specific IgE/ml. At 100–600 ng/ml Der p 2-specific IgE the level of FAP was correlated with the concentration of allergen-specific IgE, whereas at lower concentrations FAP was low or absent. All tested sera from eczema patients, all having serum anti-Der p 2-IgE concentrations > 600 ng/ml, showed a high FAP capacity, whereas all tested sera from atopic patients without eczema, which had serum anti-Der p 2-IgE levels < 600 ng/ml, showed no or a low FAP capacity. The association of high FAP capacity with eczema may reflect a functional role of FAP in the pathogenesis of atopic dermatitis.
Anti-La (SS-B) but not anti-Ro52 (SS-A) antibodies cross-react with laminin—a role in the pathogenesis of congenital heart block?
Authors: Li J-M , Horsfall A C , Maini R N
Published: July 2010

Cross-reactions between maternally derived autoantibodies and fetal cardiac antigens have been postulated to play a role in the pathogenesis of congenital heart block (CHB). We have explored the cross-reactivity of autoantibodies to the small ribonuclear autoantigens, La/SS-B and Ro/SS-A, with laminin, the major component of cardiac sarcolemmal membrane using affinity-purified antibodies from patients with Sjögren's syndrome (SS). Anti-La antibodies purified from eight of 10 patients cross-reacted significantly with mouse laminin by ELISA. In contrast, purified antibodies to Ro52 from the same 10 patients showed little or no binding to laminin. Laminin inhibited up to 70% binding of anti-La antibodies to La antigen, and La inhibited up to 65% binding of anti-La antibodies to laminin. The cross-reaction was further examined on cryosections of 10 human fetal hearts aged from 8·7 to 14·9 weeks of gestation, two normal adult hearts, and one pathological adult heart with a diagnosis of dilated cardiomyopathy. Anti-Ro52 antibodies did not bind to the surface of cardiac cells. However, anti-La antibodies from seven of 10 patients tested bound to the surface of fetal myocytes from hearts aged 9·4 to 14·9 weeks of gestation, and also to the myocytes from the pathological adult heart but not to normal adult hearts. Preincubation with La antigen abolished the binding of anti-La antibodies to the surface of adult heart myocytes with dilated cardiomyopathy, and pre-incubation with mouse laminin could partially block this binding. These results suggest that molecular mimicry between laminin and La, but not Ro52, may act as a target for specific maternal autoantibodies, and contribute to the pathogenesis of CHB at a critical stage during fetal cardiac development.
Effect ofMycoplasma arthritidissuperantigen on enzymatically induced arthritis in mice
Authors: Sustackova H , Chambers C A , Hozumi N , Kandel R A , Quinn P A , Cruz T F
Published: July 2010

The objective of this study was to examine the effect of the stimulation of the immune system with Mycoplasma arthritidis superantigen (MAS) on joint inflammation and cartilage destruction. MAS was administered either alone or combined with a model of degenerative arthritis induced by intraarticular injection of collagenase enzyme. Intraperitoneal injection of MAS resulted in activation of peripheral lymphocytes in BALB/c mice, as shown by a proliferative response of splenocytes isolated from MAS-treated animals to IL-2-containing supernatant. Intraperitoneal or intraarticular administration of MAS alone at concentrations maximally activating lymphocytes had no detectable effect on joints. Intraarticular injection of collagenase resulted in some infiltration of inflammatory cells into the joints, hyperplasia and hypertrophy of synovial lining, pannus formation and surface loss of proteoglycans 7 days following the injection. At 21 days, the animals showed almost total loss of cartilage and minimal or no inflammation. Animals receiving MAS in addition to collagenase treatment showed similar changes in the joints. These data have demonstrated that activation of the immune system with MAS in vivo does not increase joint inflammation or cartilage degradation in enzymatically induced arthritis.
Antigen-presenting cells in human cutaneous leishmaniasis due toLeishmania major
Authors: Elhassan A M , Gaafar A , Theander T G
Published: July 2010

In this study biopsies from skin lesions and draining lymph nodes of patients suffering from cutaneous leishmaniasis caused by Leishmania major were examined by immunohistochemistry, and by light and electron microscopy to identify the types of antigen-presenting cells (APC) and their location. APC, identified morphologically and by their expression of specific cell markers, included Langerhans cells, macrophages, follicular dendritic cells, and interdigitating reticulum cells of the paracortex of lymph nodes. These cells expressed MHC class II antigens and contained Leishmania antigen. Since some keratinocytes and endothelial cells also showed these characteristics, they may also act as APC. By examining tissue samples from skin lesions and draining lymph nodes it was possible to follow the probable route of trafficking of various inflammatory cells between the skin lesion and lymph nodes. Leishmania antigen containing Langerhans cells were found in the epidermis, dermis and the regional lymph nodes. We believe these cells translocate from the epidermis to the dermis, where they take up antigen and migrate to the paracortex of the regional lymph nodes. There they are intimately associated with cells of the paracortex, and could be involved in the generation of Leishmania-specific T memory cells. LFA-1 -positive T cells of the CD45RO phenotype were found in the skin lesion. Venular endothelium in the skin lesions expressed intercellular adhesion molecule-1 (ICAM-1), which is the ligand for LFA-1. The migration of lymphocytes from the vascular lumen to the site of inflammation is possibly a result of the interaction of these two adhesion molecules.
Attenuation of immune complex nephritis in NZB/WF1 mice by a prostacyclin analogue
Authors: Utsunomiya Y , Ogura M , Kawamura T , Mitarai T , Maruyama N , Sakai O
Published: July 2010

Although prostaglandins have been shown to inhibit the evolution of the nephritis in NZB/W mice, the mechanisms of this effect are unknown. To characterize such inhibition, we injected the prostacyclin (PGI2) analogue, beraprost, into NZB/W mice, using 0·5mg, 1·0mg or 5·0mg beraprost/kg body weight of test animals three times in 1 week when the mice were 2 months old. Evaluation included measurement of urine albumin excretion, serological parameters and splenic T cell subset, as well as examination of renal histology by light and fluorescence microscopy. Mice given beraprost showed a marked decrease in urine albumin excretion and in glomerular hypercellularity compared with untreated controls. Maximal beneficial effects occurred when the dose was 5·0mg/kg of beraprost. These effects correlated with a reduction of immune complex deposition in glomeruli. In addition, beraprost reduced serum levels of immunoglobulins and anti-double-stranded DNA antibodies, and decreased the number of helper (L3T4+) T cells in splenocytes. These results indicate that beraprost attenuates the nephritis of NZB/W mice, and that the source of this effect is the reduced production of autoantibodies and deposition of immune complexes in glomeruli.
Defective expression of early activation genes in cartilage-hair hypoplasia (CHH) with severe combined immunodeficiency (SCID)
Authors: Castigli E , Irani A-M , Geha R S , Chatila T
Published: July 2010

Cartilage-hair hypoplasia (CHH) is an autosomal recessive disease of unknown etiology characterized by metaphyseal dysostosis, unpigmented hair, and defective cellular immunity. We studied peripheral blood mononuclear cells (PBMC) of a boy with CHH and combined immunodeficiency in an attempt to characterize further the immune defect in this disease. Stimulation of his PBMC with mitogens was associated with severely depressed IL-2 and interferon-gamma (IFN-γ) synthesis and lL-2 receptor a-chain (IL-2Rα) expression and resulted in poor lymphocyte proliferation that was only modestly upregulated by the addition of recombinant IL-2 (rIL-2). The defective proliferation and lymphokine synthesis were not corrected by the addition of phorbol myristate acetate (PMA) and ionomycin, agents that bypass receptor-mediated signalling, indicative of a distal abnormality. Importantly, the levels of mRNA encoding c-myc, IL-2Rα, IL-2 and IFN-γ were markedly decreased in patient lymphocytes stimulated with PMA + ionomycin as compared to control lymphocytes The defect in the expression of these early activation genes was selective in that induction by mitogens of mRNA encoding other early activation gene products such as c- fos and c-jun was not impaired. These results suggest that the underlying defect in this patient and perhaps others with CHH may be an abnormality in a component of intracellular signalling pathways or in a trans-acting factor which regulates the expression of a selected number of early activation genes.