bluuebunny/crossref_metadata_2025_split huggingface dataset:
Tetanus toxoid-specific T cell responses in severe combined immunodeficiency (SCID) mice reconstituted with human peripheral blood lymphocytes
Authors: SOMASUNDARAM R , JACOB L , HERLYN D 
Published: July 2010

SUMMARYSCID mice reconstituted with human peripheral blood lymphocytes (PBL) have repeatedly been shown 10 produce antigen-specific B ceil responses. We have derived tetanus toxoid (TT)-specific human T cell lines from cells of the peritoneal cavity, spleen and lymph nodes of SCID mice reconstituted with human PBL and boosted with TT. Establishment of these cell lines was dependent on the time interval between reconstitution of the mice with human PBL and initiation of lymphocyte cultures in vitro. When lymphocytes were collected from the mice 8 weeks after reconstitution, human lymphocytes with TT-specific proliferative activity in vitro were isolated from the peritoneal cavity and spleen, but long-term cell lines could not be established after repeated lymphocyte stimulation with TT. IL-2 and autologous Epstein Barr virus-transformed B cells. In contrast, three long-term (&gt;10 months) TT-specific human T cell lines were established from lymphocytes collected from two of the eight mice in the group 4 weeks after reconstitution. The T cell lines were either CD4+ (two lines derived from peritoneal cavity and lymph node, respectively) or CD8+ (one line derived from spleen) and all expressed CD3, T cell receptor α/β, and human histocompatibility leucocyte class I antigen. The T cell lines, however, lacked cytotoxic, helper and suppressor activities. Thus, SCID mice can support human T cells that actively migrate to various organs and respond to antigenic stimuli both in vivo and in vitro, but these T cells lack characteristic functions.
Bactericidal/permeability-increasing protein (BPI) is an important antigen for anti-neutrophil cytoplasmic autoantibodies (ANCA) in vasculitis
Authors: Zhao M H , Jones S J , Lockwood C M 
Published: July 2010

SUMMARYIndirect immunofluorescence (IIF) techniques have shown that ANCA are useful serological markers for some small vessel vasculitides, and ELISA assays, using purified molecules as solid-phase ligand, have helped to identify proteinase 3 (PR3) and myeloperoxidase (MPO) as two of the major ANCA antigens. There remain a substantial number of serum samples, which are positive by IIF, yet recognize neither PR3 nor MPO (double-negative samples). We found, by Western blot analysis of soluble neutrophil granule proteins, that certain of these double-negative samples recognized a 55-kD doublet of which the first eight residues shared N-terminal amino acid sequence homology with BPI, a potent antibiotic towards Gram-negative bacteria. We developed a simple, quick and robust two-step immunobiochemical method to purify BPI. This was then employed to detect anti-BPI autoantibodies by ELISA and Western blot analysis. We tested 100 double-negative samples and 400 consecutive new samples sent for routine ANCA testing in the anti-BPI ELISA. We found that 45 of the 100 double-negative and 44 of the 400 new routine samples recognized BPI. By Western blot analysis 20/20 positive anti-BPI samples blotted the 55-kD protein. Inhibition assays confirmed the specificity of binding. Review of the 89 anti-BPI-positive patients showed a male dominance (M: F ratio 55:34), a mean age of 60.4 years and clinical diagnoses ranging from organ limited vasculitis to widespread systemic vasculitis.
IL-8/IL-8 receptor expression in psoriasis and the response to systemic tacrolimus (FK506) therapy
Authors: Lemster B H , Carroll P B , Rilo H R , Johnson N , Nikaein A , Thomson A W 
Published: July 2010

SUMMARYRecently, the keratinocyte IL-8/IL-8 receptor (IL-8R) pathway has been implicated in the pathogenesis of psoriasis, and there is evidence that the potent macrolide immune suppressant tacrolimus (formerly FK506) can inhibit this pathway in vitro. In this study, determination of the expression of cytokine mRNAs in lesional skin of patients with active disease by reverse transcriptase polymerase chain reaction revealed transcripts for IL-1β, tumour necrosis factor-alpha (TNF-α), IL-6, IL-8, IL-8R, IL-10, interferon-gamma (IFN-γ), IL-2R and transforming growth factor-beta (TGF-β), but not IL-2 or IL-4. IL-8 was the only cytokine expressed in affected skin of all patients but not in clinically normal skin of healthy subjects. In seven CD4+ T cell clones propagated from the lesional skin of an untreated psoriasis patient, IL-8 was expressed by the skin-derived T lymphocytes and not by feeder cells (irradiated autologous blood lymphocytes); IL-1β, IL-2, IL-6 and IL-10 were also expressed by some or all of the T cell clones, IL-8 mRNA was not detected in the skin of any patient after the start of systemic tacrolimus therapy; IL-lβ, IL-6 and IFN-γ transcripts were also reduced. By 12 weeks, the mean psoriasis area and severity index (PASI) had decreased from 18·8 to 3·8, a reduction of 80%. In the same post-treatment biopsies, however, message for IL-8R persisted. Estimation of circulating IL-8 levels by enzyme immunoassay showed that all patients with detectable IL-8 before treatment had decreased levels in response to treatment with tacrolimus; reductions in PASI scores were accompanied by decreases in IL-8 levels, that varied both in rate and extent. Partial relapse, which in a minority of patients followed the initial period of remission, and was precipitated by drug dose reduction, was accompanied by an increase in circulating IL-8. These findings add credence to the view that the IL-8/IL-8R autocrine/paracrine pathway may be important in the pathogenesis of psoriasis. They further suggest that interference with IL-8 production and/or that of other key chemokines may be an important mechanism underlying the therapeutic efficacy of tacrolimus, and other agents such as cyclosporin A, with similar molecular actions.
Cytokine production by normal human monocytes: inter-subject variation and relationship to an IL-1 receptor antagonist (IL-1Ra) gene polymorphism
Authors: Danis V A , Millington M , Hyland V J , Grennan D 
Published: July 2010

SUMMARYMonocytes from different individuals show variable cytokine production in response to a variety of stimuli. We wished to determine the sets of conditions (cytokine combinations) that would enable us to demonstrate stable inter-individual differences in the production of IL-1α, IL-1β, IL-1Ra, IL-6 and tumour necrosis factor-alpha (TNF-α) by monocytes. We assessed the ability of a number of recombinant human cytokines (granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon-gamma (IFN-γ), TNF-α, IL-4, IL-6, transforming growth factor-beta (TGF-β), IL-10 and IL-1Ra) to stimulate or inhibit the production of one or more of these monocyte products. GM-CSF was found to stimulate the production of all five of these cytokines in a highly reproducible manner. TNF-α also up-regulated production of IL-1α, IL-1β, IL-1Ra and IL-6 by monocytes, but the variability in the results of cells cultured from the same individuals on different occasions was greater. Other cytokines either stimulated production of only some of the five cytokine products tested, or stimulated the production of some cytokine products while inhibiting production of others. This was especially evident when cytokines were used in combination with GM-CSF: IFN-γ down-regulated production of IL-1Ra while up-regulating the production of IL-1α/β, IL-6 and TNF-α, while IL-4 had the exact opposite effect. Polymorphisms in regions of cytokine genes that affect transcription may account for some of the inter-individual variation in cytokine production. We have shown that a stable estimate of cytokine production phenotype can be obtained when monocytes collected on at least two separate occasions are stimulated by GM-CSF in vitro. We have looked for a relationship between IL-1 production and an 86-bp variable repeat polymorphism in intron 2 of the IL-1Ra gene. A less common allele of this polymorphism (allele 2) was associated with increased production of IL-1Ra protein, and also reduced production of IL-1α protein by monocytes.
Analysis of proteins that interact with the IL-2 regulatory region in patients with rheumatic diseases
Authors: BECKER H , STENGL G , STEIN M , FEDERLIN K 
Published: July 2010

SUMMARYIn order to investigate transcriptional regulation of lymphokine genes in rheumatic diseases, peripheral blood mononuclear cells from patients with systemic lupus erythematosus (SLE), rheumatoid arthritis (RA), and systemic sclerosis (SSc) were analysed for expression of DNA-binding proteins. Nuclear extracts prepared from unstimulated and mitogen-activated cells were studied for their ability to bind to 32P-labelled oligonucleotides containing the AP-1, NF-AT, NF-B and CD28RC sites of the IL-2 promoter. Using gel mobility-shift assay, detection of protein binding to the AP-1 site was reduced in SLE compared with controls. NF-AT binding activity was enhanced in all groups of patients, and was associated with measures of disease activity in RA. In addition. SSc patients showed increased NF-×B binding activity. Altered patterns of DNA-binding proteins suggest disturbed intracellular signalling which may contribute to abnormal lymphokine production in rheumatic diseases.
Rabbit polymorphonuclear granulocyte function during ethanol administration—migration and oxidative responses in a joint with immune complex synovitis
Authors: Nilsson E , Thomsen P , Ericson L , Palmblad J 
Published: July 2010

SUMMARYFunctional impairments of polymorphonuclear granulocytes (PMN) are believed to contribute to hampered inflammation and host defence in alcoholics. We studied effects of i.v. ethanol administration on PMN responses in rabbits during induction of a knee-joint synovitis. The synovitis conferred systemic effects, since chemiluminescent responses of peripheral blood PMN to opsonized zymosan and phorpbol myristate acetate (PMA) increased 64- and 17-9-fold, respectively. Chemiluminescent responses of synovial PMN were further amplified. This up-regulation was reduced to 33% in rabbits treated with ethanol when opsonized zymosan was used as the PMN stimulus: in contrast, PMA responses were unaffected. The appearance and migration of PMN to the synovitis joint were normal despite a blood ethanol concentration of 0-5%. Thus, ethanol impaired release of oxygen metabolites from PMN, but not the delivery of cells at an inflammatory site.
Detection of enteroviruses in the intestine of type 1 diabetic patients
Authors: Oikarinen M , Tauriainen S , Honkanen T , Oikarinen S , Vuori K , Kaukinen K , Rantala I , Mäki M , Hyöty H 
Published: July 2010

Summary
Enterovirus infections have been diagnosed more frequently in type 1 diabetic patients than in the healthy population, and enteroviruses have also been found in the pancreas of diabetic patients. Primary replication of the virus occurs in the gut, but there are no previous studies evaluating possible presence of virus in the intestine of diabetic patients. The purpose of this study was to investigate if enteroviruses can be found in small intestinal tissue of type 1 diabetic patients. Formalin-fixed, paraffin-embedded upper intestinal biopsy samples were analysed for the presence of enterovirus using in situ hybridization and immunohistochemistry. Enterovirus was detected by in situ hybridization in six (50%) of the type 1 diabetic patients (n = 12) but in none of the control subjects (n = 10, P = 0·015). Immunohistochemistry identified enterovirus in nine (75%) of the patients and one (10%) control subject (P = 0·004). The presence of the virus was confirmed by reverse transcription–polymerase chain reaction in one of the four patients from whom a frozen and unfixed sample was available. Intestinal morphology was normal in all study subjects. The results suggest that a substantial proportion of type 1 diabetic patients have an ongoing enterovirus infection in gut mucosa, possibly reflecting persistent enterovirus infection. This observation opens new avenues for further studies on the possible role of enteroviruses in human type 1 diabetes.
Stimulation by means of dendritic cells followed by Epstein–Barr virus-transformed B cells as antigen-presenting cells is more efficient than dendritic cells alone in inducing Aspergillus f16-specific cytotoxic T cell responses
Authors: Zhu F , Ramadan G , Davies B , Margolis D A , Keever-Taylor C A 
Published: July 2010

SummaryAdoptive immunotherapy with in vitro expanded antigen-specific cytotoxic T lymphocytes (CTLs) may be an effective approach to prevent, or even treat, Aspergillus (Asp) infections. Such lines can be generated using monocyte-derived dendritic cells (DC) as antigen-presenting cells (APC) but requires a relatively high volume of starting blood. Here we describe a method that generates Asp-specific CTL responses more efficiently using a protocol of antigen presented on DC followed by Epstein–Barr virus (EBV)-transformed B lymphoblastoid cell lines (BLCL) as APC. Peripheral blood mononuclear cells were stimulated weekly (2–5×) with a complete pool of pentadecapeptides (PPC) spanning the coding region of Asp f16 pulsed onto autologous mature DC. Cultures were split and stimulated subsequently with either PPC-DC or autologous PPC-pulsed BLCL (PPC-BLCL). Lines from the DC/BLCL arm demonstrated Asp f16-specific cytotoxicity earlier and to a higher degree than lines generated with PPC-DC alone. The DC/BLCL-primed lines showed a higher frequency of Asp f16-specific interferon (IFN)-γ producing cells but an identical effector cell phenotype and peptide specificity compared to PPC-DC-only-primed lines. Tumour necrosis factor (TNF)-α, but not IL-10, appeared to play a role in the effectiveness of BLCL as APC. These results demonstrate that BLCL serve as highly effective APC for the stimulation of Asp f16-specific T cell responses and that a culture approach using initial priming with PPC-DC followed by PPC-BLCL may be a more effective method to generate Asp f16-specific T cell lines and requires less starting blood than priming with PPC-DC alone.
99th Dahlem Conference on Infection, Inflammation and Chronic Inflammatory Disorders: Lifestyle changes affecting the host–environment interface
Authors: Ehlers S , Kaufmann S H E 
Published: July 2010

Summary
In industrialized nations and high-income regions of the world, the decline of infectious diseases is paralleled by an increase in allergic, autoimmune and chronic inflammatory diseases (AACID). Changes in lifestyle in westernized societies, which impact individually and collectively on intestinal microbiota, may – at least in part – account for the AACID pandemic. Many disease genes that contribute to AACID encode pattern recognition and signalling molecules in barrier-associated cells. Interactions between gene products and environmental factors depend highly upon the host's state of maturation, the composition of the skin and gut microflora, and exposure to pollutants, antibiotics and nutrients. Inflammatory stress responses, if regulated appropriately, ensure immunity, health and relative longevity; when they are dysregulated, they can no longer be terminated appropriately and thus precipitate AACID. The 99th Dahlem Conference brought together experts of various disciplines (genetics, evolution biology, molecular biology, structural biology, cell biology, immunology, microbiology, nutrition science, epidemiology and clinical medicine) to discuss the multi-faceted relationships between infection, immunity and inflammation in barrier organs and the development of AACID. In Clinical and Experimental Immunology we are presenting a compilation of background papers that formed the basis of discussions. Controversial viewpoints and gaps in current knowledge were examined and new concepts for prevention and treatment of CID were formulated.
99th Dahlem Conference on Infection, Inflammation and Chronic Inflammatory Disorders: Caenorhabditis elegans as a model to study tissues involved in host immunity and microbial pathogenesis
Authors: Irazoqui J E , Ausubel F M 
Published: July 2010

Summary
The molecular mechanisms involved in host–microbe interactions during the initial stages of infection are poorly understood. The bacteria-eating nematode Caenorhabditis elegans provides an opportunity to dissect host–microbe interactions in the context of the whole organism, using powerful genomic, genetic and cell-biological tools. Because of the evolutionary conservation of ancient innate host defences and bacterial virulence mechanisms, studies in C. elegans hold great promise to shed light on defences in higher organisms, including mammals. Additionally, C. elegans pathogenesis models provide a platform for the identification of novel classes of anti-infective compounds with therapeutic value.
Revealing secret liaisons: DNA barcoding changes our understanding of food webs
Authors: KAARTINEN RIIKKA , STONE GRAHAM N. , HEARN JACK , LOHSE KONRAD , ROSLIN TOMAS 
Published: July 2010

1. How food webs are structured will affect how species dynamically interact. To date, the construction of quantitative food webs has largely been based on morphological species characters. Yet, recent work suggests that the use of molecular characters may change our perception of both species limits and species identity.2. Focusing on an assemblage of natural enemies of specialist herbivores, we used DNA sequence ‘barcode’ identification for three purposes: to screen the focal web for morphologically cryptic species, to catch misidentified individuals and to re‐examine key descriptors of food web structure.3. Based on morphological characters, we were able to distinguish 51 parasitoid species and 5 inquilines. Molecular markers revealed four morphologically cryptic parasitoid taxa and completely changed our view of species limits among the inquilines, defining six molecular operational taxonomic units (MOTUs) with little resemblance to the original five morphospecies.4. In total, the use of DNA barcode information changed the species designation of 31% of the sequenced individuals. However, measures of food web structure changed relatively little, reflecting the fact that the cryptic taxa detected by us accounted for only a minor proportion of interactions in the overall food web. Qualitatively, web metrics suggested higher specialisation of parasitoid species in molecularly informed than in purely morpohologically‐based webs.5. Taken together, this study suggests that the added resolution offered by molecular information contributes a new level of precision to food web studies. Overall, it depicts interactions within the food web as more specialised than revealed by morphological species characters. Importantly, molecular markers will also allow us to match morphologically cryptic taxa sampled among regions, and thereby to compare the ecological role of individual species in different communities.
Will climate change reduce the effects of a pesticide on amphibians?: partitioning effects on exposure and susceptibility to contaminants
Authors: ROHR JASON R. , SESTERHENN TIMOTHY M. , STIEHA CHRISTOPHER 
Published: July 2010

AbstractSeveral studies suggest that global climate change could increase the toxicity of contaminants, but none of these studies explicitly integrate the effects of climate change on both susceptibility and duration of exposure to pollution. For many amphibian and aquatic insect species, exposure to contaminants is probably greatest during their fully aquatic embryonic and larval stages because these stages cannot readily escape water bodies where many contaminants accumulate and concentrate. Hence, by accelerating embryonic and larval development, global warming might reduce the duration of contaminant exposure for these taxa. To test this hypothesis, we isolated the effects of a temperature gradient (13–25 °C) on susceptibility (toxicity at a controlled exposure duration) and exposure of the streamside salamander, Ambystoma barbouri, to the herbicide atrazine (0, 4, 40, and 400 μg L−1) by quantifying growth, survival, hatching, and metamorphosis under an atrazine exposure duration that was either constant or that depended on time to metamorphosis (and thus temperature). Increasing atrazine concentrations reduced growth, delayed hatching and metamorphosis, and decreased embryonic and larval survival. Increasing temperatures enhanced growth, accelerated development, and reduced survival for embryos but not larvae. With the exception of growth, increasing temperatures generally did not enhance the toxicity of atrazine, but they did generally ameliorate the adverse effects of atrazine by accelerating development and reducing the duration of atrazine exposure. The actual effects of climate change on contaminants remains difficult to predict because temperature changes can affect chemical use, uptake, excretion, biotransformation, fate, transport, and bioavailability. However, this work highlights the importance of explicitly considering how climate change will affect both exposure and toxicity to contaminants to accurately assess risk.
Fell‐Muir Lecture: Metalloproteinases: from demolition squad to master regulators
Authors: Murphy Gillian 
Published: July 2010

SummaryTwo families of the Metzincin clan of metalloproteinases, the matrix metalloproteinases and the disintegrin metalloproteinases have attracted much attention as important effectors of cellular interactions with their environment. They appear to play significant roles in the modulation of components of the extracellular matrix, matrix and cell receptors, as well as the cytokines and growth factors and their receptors. Such functions at the ‘cutting edge’ of cell biology puts these enzymes in pivotal roles in the orchestration of the rapid response of cells to their environment, acting as key switches between different signalling pathways. Inevitably such enzymes should be regarded as suitable targets for therapeutic approaches to many diseases where such pathways become dysregulated. A major challenge to the development of direct inhibitors of catalysis has been the broad structural similarity of the Metzincin catalytic site. More detailed knowledge of active site structures has helped to some extent to resolve the development of more specific chemical inhibitors and selected enzymes are now being targeted. An alternative strategy is the consideration of the role of the extracatalytic domains that are determinants of specificity at a variety of levels. Dissecting the relationships between structure and function of these interaction sites is allowing the development of new approaches to inhibition of enzyme function. Antibodies are proving useful tools in this respect and may pave the way to a novel biologics approach to disease therapy.
An electron microscopical investigation of fat destabilization in ice cream
Authors: BERGER K. G. , WHITE G. W. 
Published: July 2010

SummaryA limited amount of fat destabilization occurs when ice cream mix is aerated and frozen, and this emulsion breakdown imparts some desirable properties to ice cream. A method of measuring the amount of de‐emulsified fat is described, and the results of an investigation of the process by means of electron microscopy are given. The effects of fat composition and processing variables on fat destabilization are summarized, and an overall picture is presented of the part played by fat in the structure of ice cream.