bluuebunny/crossref_metadata_2025_split huggingface dataset:
A Recombinant Vaccinia Virus Encoding the Interferon‐Inducible T‐Cell Alpha Chemoattractant is Attenuated In Vivo
Authors: Hamilton N. H. R. , Mahalingam S. , Banyer J. L. , Ramshaw I. A. , Thomson S. A. 
Published: March 2004

AbstractMurine interferon‐inducible T‐cell alpha chemoattractant (I‐TAC) is a potent non‐ELR CXC chemokine that predominantly attracts activated T lymphocytes, binds to the receptor CXCR3 and is induced by interferon‐γ (IFN‐γ). We analysed I‐TAC expression by reverse transcriptase‐polymerase chain reaction during three different virus‐infection models in mice, respiratory syncytial virus (RSV), influenza A and vaccinia virus western reserve (VV‐WR). In the lungs from mice infected with RSV or influenza A viruses, peak expression of I‐TAC coincided with peak viraemia. Surprisingly, there was no expression in the lungs of mice infected with vaccinia, unlike the elevated expression shown previously for other interferon‐regulated chemokines, such as Crg2 and Mig. To further investigate the importance of this difference during vaccinia infection in mice, a recombinant virus encoding I‐TAC (rVV I‐TAC) was generated. Studies in C57BL/6 and Swiss nude mice showed that I‐TAC expression caused increased mononuclear cell infiltration and significantly attenuated the VV. Infection of the footpads of naïve or already immune (with VV‐WR) mice with either rVV I‐TAC or VV‐WR demonstrated that I‐TAC expression reduced overall inflammation during infection and that this reduction was more pronounced in already immune mice. The data presented here show that I‐TAC can have an important role during virus infections and that vaccinia has evolved ways to avoid inducing I‐TAC expression.
Regulation of Mast Cell Migration by TH1 and TH2 Cytokines: Identification of Tumour Necrosis Factor‐α and Interleukin‐4 as Mast Cell Chemotaxins
Authors: Olsson N. , Taub D. D. , Nilsson G. 
Published: March 2004

AbstractMast cells act as central effector and regulatory cells in many inflammatory disorders, including T helper 1 (TH1)‐mediated inflammations such as autoimmunity and TH2‐mediated inflammations such as allergy and parasite infections. One characteristic for mast cell‐mediated inflammations is the accumulation of mast cells in the inflamed tissue. The factors regulating mast cell recruitment in these inflammations are still not fully characterized. We have investigated the potency of TH1‐ and TH2‐secreted cytokines to mediate mast cell migration. Supernatants from six different TH1 and TH2 clones were tested for mast cell‐chemotactic activity using the human mast cell line (HMC‐1) as a responder cell. All six clones produced factors that induced mast cell migration. Using blocking antibodies to a broad range of cytokines, we found that anti‐tumour necrosis factor‐α (anti‐TNF‐α) reduced the migration of mast cells to supernatants from TH1 clones. In contrast, the main mast cell chemoattractants secreted by TH2 clones were found to be interleukin‐4 (IL‐4) and IL‐8. The potency of these cytokines to act as mast cell chemoattractants was confirmed by using recombinant IL‐4, IL‐8 and TNF‐α. Our results suggest that TNF‐α can be involved in the recruitment of mast cells in TH1‐mediated inflammations, whereas IL‐4 and IL‐8 might play a similar role in TH2‐mediated inflammations.
Antibodies to GAD65 Epitopes at Diagnosis and Over the First 10 years of Clinical Type 1 Diabetes Mellitus
Authors: Ronkainen M. S. , Savola K. , Knip M. 
Published: March 2004

AbstractAntibodies to glutamate decarboxylase (GAD65Ab) may persist, and their titres even increase after the clinical onset of type 1 diabetes. To characterize this phenomenon in detail, we analysed sequentially antibodies to GAD65 epitope clusters in a radio‐binding assay in patients with type 1 diabetes. Serum samples were taken at diagnosis and 2, 5 and 10 years later from 50 young patients who had tested positive for GAD65Ab at least once during observation. The levels of GAD65Ab peaked in 21 patients after diagnosis. Antibodies to the middle region of GAD65 (GAD65‐M‐Ab, 88%) were more common at diagnosis than antibodies to the C‐terminal (GAD65‐C‐Ab, 68%, P < 0.05) or N‐terminal region (4%, P < 0.001). Antibodies to middle and especially to C‐terminal epitopes decreased in those with decreasing levels of GAD65Ab (P < 0.001), whereas the frequencies of GAD65‐M‐Ab and GAD65‐C‐Ab remained quite stable among the subjects with increasing levels. Lower exogenous insulin dose and HbA1 levels and stronger humoral immune response to islet cells were observed in those with increasing levels of GAD65‐M‐Ab than in those with decreasing levels (P < 0.05). The present observation supports the view that the middle region of GAD65 comprises immunodominant epitopes. An enhanced humoral immune response to GAD65 after diagnosis is related to persistent immune reactivity to the middle and C‐terminal regions.
Severe Hypercholesterolaemia Leads to Strong Th2 Responses to an Exogenous Antigen
Authors: Robertson A.‐K. L. , Zhou X. , Strandvik B. , Hansson G. K. 
Published: March 2004

AbstractSevere hypercholesterolaemia is associated with decreased levels of immunoglobulin G2a (IgG2a) antibodies [T‐helper 1 (Th1) response] to modified malondialdehyde‐modified low‐density lipoprotein (MDA‐LDL) and increased levels of Th2‐dependent IgG1 antibodies in apolipoprotein E‐deficient (apoE–/–) mice. To investigate whether this reflects a general pattern of metabolic regulation of the humoral immune response, apoE–/– mice were fed diets resulting in different degrees of hypercholesterolaemia and immunized with keyhole limpet haemocyanin (KLH) in aluminium hydroxide. Cholesterol levels for different treatment groups ranged from 14 to 77 mmol/l in serum and from 10 to 39 mmol/g in liver. Mice with severe hypercholesterolaemia had increased IgG1 antibodies to MDA‐LDL and decreased IgG2a anti‐MDA‐LDL. Importantly, titres of IgG2a antibodies to KLH were also decreased, while IgE anti‐KLH was increased, with a corresponding induction of interleukin‐4 (IL‐4) and IL‐10 and a decrease in interferon‐γ (IFN‐γ) in KLH‐stimulated spleen cells in vitro. Thus, hypercholesterolaemia clearly affects antibody production both to the autoantigen MDA‐LDL and to the exogenous antigen KLH, favouring antibody isotypes (IgG1 and IgE) that are dependent on Th2 help to B cells. Nuclear receptors ligated by oxidized lipid derivatives modulate T‐cell responses, and it is speculated that this mechanism may cause the switch to Th2 in severe hypercholesterolaemia.
Exposure to continuous bromodeoxyuridine (BrdU) differentially affects cell cycle progression of human breast and bladder cancer cell lines
Authors: Diermeier S. , Schmidt‐Bruecken E. , Kubbies M. , Kunz‐Schughart L. A. , Brockhoff G. 
Published: March 2004

Abstract.   Incorporation of bromodeoxyuridine (BrdU) during DNA replication is frequently used for cell cycle analysis. The flow cytometric BrdU/Hoechst quenching technique is conducive to high‐resolution assessment of cell cycle kinetics, but requires continuous BrdU treatment, which may have cytostatic or cytotoxic effects. Here, we have examined the impact of BrdU on the proliferation of BT474 and SK‐BR‐3 breast cancer cell lines and compared the observed effects with cell proliferation of RT4 and J82 bladder carcinoma cells, previously described to be sensitive and insensitive to BrdU, respectively. Both uni‐ and bi‐parametric DNA measurements were performed to identify BrdU‐induced alterations in the S‐phase fraction and in cell cycle progression. An annexinV/propidium iodide (PI) assay was used to identify potential induction of apoptosis by BrdU. Proliferative activity in BT474, SK‐BR‐3, and RT4 cultures was reduced in different cell cycle phases due to continuous treatment with 60, 5.0, and 3.5 µm BrdU. This effect, which was not found in J82 cultures, was dependent on exposure time (96 versus 48 h) and was also dose‐dependent for RT4 and SK‐BR‐3. BrdU application does not induce apoptosis or necrosis as revealed with the annexin V/PI assay. We concluded that continuous BrdU treatment did not affect cell viability, but essentially alters cell cycle progression in three out of four cell lines tested. Cell‐type specific validation of the feasibility of the powerful BrdU/Hoechst quenching technique is required and recommended.
Blocking lymphotoxin-βreceptor activation diminishes inflammation via reduced mucosal addressin cell adhesion molecule-1 (MAdCAM-1) expression and leucocyte margination in chronic DSS-induced colitis
Authors: Stopfer P , Obermeier F , Dunger N , Falk W , Farkas S , Janotta M , Möller A , Männel D N , Hehlgans T 
Published: March 2004

SUMMARYThe lymphotoxin-β receptor (LTβR) pathway is critical for maintenance of organized lymphoid structures and is involved in the development of colitis. To investigate the mechanisms by which LTβR activation contributes to the pathology of chronic inflammation we used a soluble LTβR-Ig fusion protein as a competitive inhibitor of LTβR activation in the mouse model of chronic colitis induced by oral administration of dextran sulphate sodium. Strong expression of LTβ which constitutes part of the LTα1β2 ligand complex was detected in colonic tissue of mice with chronic colitis. Treatment with LTβR-Ig significantly attenuated the development and histological manifestations of the chronic inflammation and reduced the production of inflammatory cytokines such as TNF, IL-1β, and IL-6. Moreover, LTβR-Ig treatment significantly down-regulated mucosal addressin cell adhesion molecule-1 (MAdCAM-1) expression, leading to reduced leucocyte rolling and sticking in postcapillary and collecting venules and reduced extravasation into the intestinal mucosa as quantified by in vivo fluorescence microscopy. Thus, LTβR pathway inhibition ameliorates DSS-induced experimental chronic colitis in mice by MAdCAM-1 down-regulation entailing reduced lymphocyte margination and extravasation into the inflamed mucosa. Therefore, a combined treatment with reagents blocking T cell-mediated perpetuation of chronic inflammation such as LTβR-Ig together with direct anti-inflammatory reagents such as TNF inhibitors could constitute a promising treatment strategy for chronic colitis.
Maternal immunity to insulin does not affect diabetes risk in progeny of non obese diabetic mice
Authors: KOCZWARA K , ZIEGLER A-G , BONIFACIO E 
Published: March 2004

SUMMARYIt has been suggested that maternal environment, in particular maternal autoantibodies, modify the risk of developing autoimmune diabetes in offspring. The aim of this study was to determine whether modification of maternal environment and maternal diabetes risk through immunization affects autoimmune diabetes risk in the progeny. The risk of developing insulin antibodies and of developing diabetes was determined in 113 female progeny of non obese diabetic (NOD) dams that were immunized with insulin, control antigen or vehicle before or during pregnancy. Although NOD dams immunized with insulin were rendered diabetes resistant (40% diabetes by age 36 weeks versus 100% in control dams), diabetes development in their female offspring (72%, 26/36) was similar to that in female offspring of dams immunized with glucagon (82%, 22/27) or vehicle (76%, 19/25). Furthermore, no significant differences in diabetes development or insulin autoantibody titres were observed between female progeny of insulin autoantibody positive NOD dams (82% diabetes by age 36 weeks, 18/22), insulin autoantibody negative NOD dams (75%, 41/55), and NOD dams that had antibodies against exogneous insulin (71%, 22/31). The findings suggest that modification of the maternal risk for autoimmune diabetes via antigen-specific immunization is not transferred to progeny and that fetal exposure to insulin autoantibodies does not increase the risk for diabetes development.
Binding capacity and pathophysiological effects of IgA1 from patients with IgA nephropathy on human glomerular mesangial cells
Authors: Wang Y , Zhao M-H , Zhang Y-K , Li X-M , Wang H-Y 
Published: March 2004

SUMMARYIgA deposition in glomerular mesangium and the interaction with mesangial cells may well be the final common pathway to IgA nephropathy (IgAN). Altered hinge-region O-glycosylation of IgA1 from patients with IgAN may predispose to mesangial deposition and activation of the mesangial cell (MC) by IgA1, via a novel IgA1 receptor, and may be a key event in the pathogensis of IgAN. The aim of this study was to investigate the binding capacity and biological effects of IgA1, from both patients with IgAN and healthy controls, on human mesangial cells (HMC). Serum IgA1 was isolated with jacalin affinity chromatography, heated to aggregated form (aIgA1) and labelled with 125I. Binding capacity of aIgA1 in vitro to cultured primary HMC was evaluated by a radioligand binding assay and the specificity of binding was determined by a competitive inhibition assay. Intracellular calcium release was studied by confocal analysis and phosphorylation of extracellular signal-regulated kinase (ERK) was determined by Western blot analysis. Change of cell cycles was demonstrated by flow cytometry and HMC proliferation was evaluated by direct cell count. Expression of TGF-β mRNA and production of supernatant fibronectin were tested by RT-PCR and indirect competitive ELISA, respectively. aIgA1 from both the patients with IgAN and normal controls bound to HMC in a dose-dependent, saturable manner, and was saturated at approximately 500 pmoles per 0·5 ml of aIgA1. aIgA1 from patients with IgAN, however, bound to HMC at a higher speed and Scatchard analysis revealed a Kd of (8·89 ± 2·1) × 10−8mversus (4·3 ± 1·2) × 10−7m for aIgA1 from healthy controls (P = 0·026). The binding was specific because it was only inhibited by unlabelled Mono-IgA1 (mIgA1) and not by serum albumin or IgG. aIgA1 from patients with IgAN could induce release of intracellular calcium, phosphorylation of ERK, DNA synthesis, proliferation of HMC, expression of TGF-βmRNA and secretion of fibronectin in HMC in a similar time-dependent manner as aIgA1 from healthy controls, but the effects were much stronger and the durations were much longer (P &lt; 0·05, respectively). We conclude that aIgA1 from patients with IgAN has a higher binding capacity to HMC and stronger biological effects than aIgA1 from healthy controls. This suggests that direct interaction between IgA1 and HMC and subsequential pathophysiological responses may play an important role in the pathogenesis for IgAN.
Increased risk of adverse events when changing intravenous immunoglobulin preparations
Authors: AMERATUNGA R , SINCLAIR J , KOLBE J 
Published: March 2004

SUMMARY
Intravenous immunoglobulin (IVIG) therapy has represented a major advance in the treatment of patients with primary immune deficiency disorders. In September 2000 a new IVIG formulation, Intragam P, was introduced into clinical use. Intragam P is prepared by delipidation of pooled plasma followed by an ion exchange chromatography step to eliminate immunoglobulin aggregates. It is then pasteurized for 10 h at 60°C for viral inactivation before storage at pH 4·25 in 10% maltose. We report initial clinical experience with this new preparation. The details of adverse reactions of patients who received the new preparation were gathered shortly after it became apparent there was a change in IVIG formulation. Seven of 49 patients receiving Intragam P spontaneously reported adverse effects, which were temporally related to infusions. Subsequently, all seven patients have been able to tolerate the product with prophylactic use of antihistamines and paracetamol. This case series indicates that long-term tolerance of an older IVIG product does not necessarily equate to tolerance to a newer product, even if technically superior. Caution should be exercised when changing IVIG products, as they are not biologically equivalent.
Serum granulysin is a marker for Th1 type immunity in pre-eclampsia
Authors: Sakai M , Ogawa K , Shiozaki A , Yoneda S , Sasaki Y , Nagata K , Saito S 
Published: March 2004

SUMMARYRecent studies suggest that pre-eclampsia is associated with a Th1 predominant state and may be considered a failure of tolerance. Granulysin is a cytotoxic granule protein of natural killer (NK) cells and cytotoxic T lymphocytes (CTLs). Recently, we developed an enzyme-linked immunosorbent assay (ELISA) system for detecting serum granulysin, and reported that serum granulysin is a useful marker to evaluate the cell-mediated immunity. In this study, we show that the serum levels of granulysin were significantly elevated in pre-eclamptic patients compared with those in normal pregnancy subjects. In addition, the serum granulysin levels in pre-eclamptic patients were well associated with mean blood pressure, percentage of peripheral blood Th1 cells and Th1/Th2 ratios. The present results suggest that the serum granulysin levels would be a useful and novel serum marker to evaluate the Th1/Th2 balance, especially Th1 type immunity in pre-eclampsia.
Plasma inflammatory cytokines and chemokines in severe acute respiratory syndrome
Authors: WONG C K , LAM C W K , WU A K L , IP W K , LEE N L S , CHAN I H S , LIT L C W , HUI D S C , CHAN M H M , CHUNG S S C , SUNG J J Y 
Published: March 2004

SUMMARY
Severe acute respiratory syndrome (SARS) is a recently emerged infectious disease caused by a novel coronavirus, but its immunopathological mechanisms have not yet been fully elucidated. We investigated changes in plasma T helper (Th) cell cytokines, inflammatory cytokines and chemokines in 20 patients diagnosed with SARS. Cytokine profile of SARS patients showed marked elevation of Th1 cytokine interferon (IFN)-γ, inflammatory cytokines interleukin (IL)-1, IL-6 and IL-12 for at least 2 weeks after disease onset, but there was no significant elevation of inflammatory cytokine tumour necrosis factor (TNF)-α, anti-inflammatory cytokine IL-10, Th1 cytokine IL-2 and Th2 cytokine IL-4. The chemokine profile demonstrated significant elevation of neutrophil chemokine IL-8, monocyte chemoattractant protein-1 (MCP-1), and Th1 chemokine IFN-γ-inducible protein-10 (IP-10). Corticosteroid reduced significantly IL-8, MCP-1 and IP-10 concentrations from 5 to 8 days after treatment (all P &lt; 0·001). Together, the elevation of Th1 cytokine IFN-γ, inflammatory cytokines IL-1, IL-6 and IL-12 and chemokines IL-8, MCP-1 and IP-10 confirmed the activation of Th1 cell-mediated immunity and hyperinnate inflammatory response in SARS through the accumulation of monocytes/macrophages and neutrophils.
GAD65 antibody isotypes and epitope recognition during the prediabetic process in siblings of children with type I diabetes
Authors:   THE CHILDHOOD DIABETES IN FINLAND STUDY GROUP, HOPPU S , RONKAINEN M S , KULMALA P , ÅKerblom H K , KNIP M 
Published: March 2004

SUMMARYWe observed 42 initially non-diabetic siblings of affected children to characterize the humoral immune response to the 65 kDa isoform of glutamic acid decarboxylase (GAD65) in preclinical type I diabetes. During the observation period with a mean duration of 9·6 years 21 siblings progressed to type I diabetes. The humoral immune response to GAD65 was observed initially as a simultaneous response to the middle (M) and carboxy (C)-terminal regions of the GAD65 molecule in most cases, and if the response was restricted initially to the middle region, it spread rapidly to the C-terminal domain and in a few cases later to the amino (N)-terminal domain. There was some heterogeneity in the GAD65 isotype response, but it was composed mainly of antibodies of immunoglobulin (Ig) G1 subclass. Responses of IgG2-, IgG4-, IgM- and IgA-GAD65Ab were observed frequently, whereas IgE- and IgG3-GAD65Ab responses were seen more rarely. Initially, the non-progressors tended more often to have IgG2- and IgG4-GAD65Ab than the progressors. As a sign of a dynamic process a significant isotype spreading was seen for IgG2-GAD65Ab (P &lt; 0·05) and close to significant for IgM (P = 0·06) among progressors and for IgM-GAD65Ab (P &lt; 0·05) among non-progressors during the observation period. This study failed to identify any GAD65 epitope- or isotype-specific antibody reactivity that could be used as a marker for progression to disease, as such progression was not associated with any specific changes in reactivity over time. Our findings indicate that epitope- and isotype-specific GAD65 antibodies are hardly capable of separating progressors from non-progressors among GAD65Ab-positive first-degree relatives of children with type I diabetes.
Interleukin-6 production and secretion by human parathyroids
Authors: SAFLEY S A , VILLINGER F , JACKSON E H , TUCKER-BURDEN C , COHEN C , WEBER C J 
Published: March 2004

SUMMARYParathyroid hormone (PTH) stimulates osteoblasts to produce the proinflammatory cytokine interleukin-6 (IL-6), causing bone resorption. In patients with primary hyperparathyroidism, elevated serum levels of IL-6 normalize after resection of parathyroid tumours. Because IL-6 is also expressed in normal parathyroids and in other endocrine cells (adrenal and islet), we hypothesized that parathyroid tumours might contribute directly to the elevated serum IL-6 levels in patients with hyperparathyroidism. Immunohistochemistry identified IL-6, PTH, and chromogranin-A (an endocrine and neuroendocrine tumour marker) in normal, adenomatous and hyperplastic parathyroids. Using immunofluorescence and confocal microscopy, IL-6 co-localized with PTH and with chromogranin-A in parathyroid cells. All cultured parathyroid tumours secreted IL-6 at levels markedly higher than optimally stimulated peripheral blood mononuclear cells. Supernates from cultured parathyroids stimulated proliferation of an IL-6-dependent cell line, and anti-IL-6 MoAb abolished this stimulatory effect. IL-6 mRNA was documented in cultured parathyroid tumours, cultured normal parathyroids, fresh operative parathyroid tumours and fresh operative normal specimens. In conclusion, these data show that parathyroid tumours and normal parathyroids contain, produce and secrete IL-6. Our findings present a novel pathway by which human parathyroids may contribute markedly to IL-6 production and elevation of serum IL-6 levels in patients with hyperparathyroidism. The physiological relevance of IL-6 production by human parathyroids remains to be determined, but IL-6 secretion by parathyroid tumours may contribute to bone loss and to other multi-system complaints observed in these patients.