bluuebunny/crossref_metadata_2025_split huggingface dataset:
The common heat shock protein receptor CD91 is up-regulated on monocytes of advanced melanoma slow progressors
Authors: STEBBING J , BOWER M , GAZZARD B , WILDFIRE A , PANDHA H , DALGLEISH A , SPICER J 
Published: October 2004

SUMMARYDespite advances in our understanding of tumour immunology there is no therapy of proven survival benefit for advanced melanoma. Nevertheless, disease progression is slow in a small proportion of patients with metastatic melanoma, suggesting a contribution to outcome from host factors. Recent data have indicated the importance of the heat shock protein receptor CD91 in immune responses to, and progression of, infectious disease. Here we investigate the relationship between CD91 expression and outcome in malignancy. Rare melanoma patients were recruited with advanced disease that was progressing unusually slowly. CD91 expression on their monocytes was compared with control patients with more typical rapidly advancing metastatic disease. Th1 and Th2 cytokines, as well as innate and adaptive immune subsets, were also measured in the two groups. A significant increase in median CD91 expression levels was observed in slow progressors (P = 0·006). There were no differences in other immune subset markers or inflammatory cytokines. The ability of CD91 to internalize and cross-present tumour antigens through the major histocompatibility complex class I pathway may maintain CD8-positive cytotoxic T cell responses and contribute to slow progression of advanced melanoma.
Allogenic donor splenocytes pretreated with antisense peptide against B7 prolong cardiac allograft survival
Authors: CHEN J , HE Q , ZHANG R , CHU Y , WANG Y , LIU Q , XIONG S 
Published: October 2004

SUMMARYThe interaction of T cell CD28/CTLA-4 receptors with B7 on antigen-presenting cells (APCs) represents an important co-stimulatory pathway in T cell activation or anergy. Our previous study indicated that recipients immunized with allogenic donor immature dendritic cells (DCs) or resting B cells could induce specific immune tolerance and prolong allograft survival. A possible mechanism for this observation is that the expression of B7 molecules is either at a low level or lacking on these cells. The present study investigates whether blockade of B7 molecules on donor splenocytes with a B7 antisense peptide (B7AP), i.e. a peptide analogue of the CD28-binding region, could induce specific immune tolerance and prolong allograft survival in the recipients. Both the lymphocyte proliferation reaction and the mice pinna cardiac allograft experiment were performed to evaluate the role of B7AP in inducing specific immune tolerance in recipients in vitro and in vivo. The results showed that 56·65% and 20·52% of C57BL/6 splenocytes expressed B7.1 and B7.2 molecules, respectively, on their cell surface. There were no significant changes of the B7 expression on such splenocytes after being treated by the B7AP (53·28% and 19·06%, respectively). B7AP inhibited the mixed lymphocyte reaction by up to 38·4% and a dose-response correlation was observed for inhibition. The recipients (BALB/c) immunized with B7AP-pretreated C57BL/6 splenocytes induced a specific immune hypo-response (43%versus control) and notably prolonged survival of the C57BL/6 cardiac allograft by up to 20·3 days. In contrast to the normal saline group (average: 8·6 days) and FTD10 control peptide group (&lt;4 days), the cardiac allograft survival of the test group was extended for an additional 11·7 days. These results strongly support the notion that immunization with donor splenocytes, which had been pretreated with B7AP, induced specific immune tolerance and prolonged allograft survival in the recipients.
Gene expression analysis of peripheral T cells in a subgroup of common variable immunodeficiency shows predominance of CCR7– effector-memory T cells
Authors: HOLM A M , SIVERTSEN E A , TUNHEIM S H , HAUG T , BJERKELI V , YNDESTAD A , AUKRUST P , FRØLAND S S 
Published: October 2004

SUMMARYCommon variable immunodeficiency (CVID) represents a heterogeneous group of antibody deficiency syndromes, characterized by defective antibody production in which T cell deficiency may play a pathogenic role. A subgroup of CVID patients has impaired in vitro T cell proliferation. Using microarray analyses of T cells from these patients, we found a gene expression pattern different from healthy controls and patients with X-linked agammaglobulinaemia. The profile of the differentially expressed genes suggests enhanced cytotoxic effector functions, antigen experienced or chronically activated T cells and a predominance of CCR7– T cells. Further experiments using flow cytometry revealed a striking predominance of CCR7– T cells in a subgroup of CVID patients, and an association with impaired T cell proliferation. Our observations indicate that a predominance of CCR7– T cells with effector-memory cell features and with reduced proliferative capacity may characterize a subgroup of CVID.
Interleukin‐15 is not required for the induction or maintenance of orally induced peripheral tolerance
Authors: Millington Owain R. , Wei Xiao‐Qing , Garside Paul , Mowat Allan MC I. 
Published: October 2004

SummaryOrally induced tolerance is a physiologically relevant form of peripheral tolerance, which is believed to be important for the prevention of pathological immune responses in the gut. Of several mechanisms proposed to mediate oral tolerance, one that has received much attention recently is the concept of regulatory CD4+ T cells. As recent studies have suggested that interleukin (IL)‐15 may be important for the differentiation and maintenance of regulatory CD4+ T cells, we have examined the role of IL‐15 in oral tolerance, using a soluble form of the IL‐15 receptor (sIL‐15R) which blocks the biological effects of IL‐15 in vivo. Oral tolerance induced by feeding mice ovalbumin (OVA) in a low‐dose regimen believed to induce regulatory T cell activity was not affected by the administration of sIL‐15R during either the induction or maintenance phase of tolerance. Thus, oral tolerance does not involve an IL‐15‐dependent mechanism.
Ovine dendritic cells transduced with an adenoviral CTLA4eEGFP fusion protein construct induce hyporesponsiveness to allostimulation
Authors: Newland Ashley , Kireta Svjetlana , Russ Graeme , Krishnan Ravi 
Published: October 2004

SummaryCTLA4 (CD152) is a transmembrane molecule expressed on activated T cells and functions as a negative regulator of T cell activation upon binding to the costimulatory molecules CD80/86. In this study, CTLA4eEGFP constructs were engineered by cloning the extracellular domains of ovine and human CTLA4 (CTLA4e) ‘in frame’ with the enhanced green fluorescent protein (EGFP). Recombinant adenoviral vectors were generated by incorporation of the CTLA4eEGFP sequence into the adenoviral genome using homologous recombination in Esherichia coli. The functional activity of the adenoviral vectors was shown by the secretion of the CTLA4eEGFP upon infection of ovine fibroblasts and the binding of the fusion protein to the target ovine and human dendritic cells expressing CD80/86 receptors by flow cytometry. The EGFP tag facilitated molecular size determinations and quantification of the secreted ovine CTLA4 fusion protein by immunoprecipitation and enzyme‐linked immunosorbent assay (ELISA), respectively, using anti‐GFP mAbs. Ovine dendritic cells obtained from pseudoafferent lymphatic cannulation of sheep were characterized based on high major histocompatibility complex (MHC) class II expression and cross‐reactivity with monoclonal antibodies to the human dendritic cell markers, CD83 and CMRF‐56. In addition, ovine dendritic cells (DC) were transfected with the adenoviral CTLA4eEGFP and when used as stimulators in a mixed lymphocyte reaction showed a reduced capacity to induce allogeneic lymphocyte proliferation. This study verifies that the ovine CTLA4eEGFP fusion protein functions similarly to its human homologue and that DC modified with adenoviral CTLA4‐EGFP may provide an effective therapeutic approach in targeting alloreactive T cells to prolong allograft acceptance in a preclinical ovine model of renal transplantation.
Transcription of major histocompatibility complex class I (Kb) and transporter associated with antigen processing 1 and 2 genes is up‐regulated with age
Authors: Assounga Alain G. , Warner Carol M. 
Published: October 2004

SummaryThe transporter associated with antigen processing 1 and 2 (TAP1 and TAP2) genes belong to the ATP‐binding cassette family of transporter genes. They provide peptides necessary for the assembly of major histocompatibility complex (MHC) class I molecules by transporting these peptides into the endoplasmic reticulum. As MHC class I protein expression increases with age, we have explored the effect of age on the transcription of MHC class I genes (Kb) and TAP1 and TAP2 genes in C57BL/6 mice. Blood and spleen lymphocytes were isolated from mice aged from 3 months to over 24 months. RNA was extracted and mRNA for Kb, TAP1, TAP2 was quantified using slot‐blot hybridization followed by densitometry. There was a parallel age‐related increase (1·5‐fold) in blood lymphocyte mRNA of these genes from 3 months to 21 months. In mice over 24 months old there was a decrease in Kb and TAP1 mRNA, but an increase in TAP2 mRNA. In spleen lymphocytes an age‐related increase in all three mRNA species occurred throughout life. While MHC class I and Tap genes underwent very similar age‐related changes, MHC class I mRNA was about 50 times more abundant than either TAP1 or TAP2 mRNA.
Interleukin‐10 secreted by B‐1 cells modulates the phagocytic activity of murine macrophages in vitro
Authors: Popi Ana Flavia , Lopes José Daniel , Mariano Mario 
Published: October 2004

SummaryAs demonstrated previously in our laboratory, B‐1 cells migrate from the peritoneal cavity of mice and home to a distant site of inflammation to become macrophage‐like cells. However, the influence that these cells might have on the kinetics and fate of the inflammatory process is not known. Considering that macrophages are pivotal in the inflammatory reaction, we decided to investigate the possible influence B‐1 cells could have on macrophage activities in vitro. Our results show that peritoneal macrophages from Xid mice, a mouse strain deprived of B‐1 cells, have higher phagocytic indexes for zymozan particles when compared with macrophages from wild‐type mice. Moreover, macrophages from wild‐type mice have a lower ability to release nitric oxide and hydrogen peroxide when compared with macrophages from Xid mice. Experiments using cocultures of B‐1 cells and macrophages from Xid mice in transwell plates demonstrated that B‐1 cells down‐regulate macrophage activities. These observations also indicate that this phenomenon is not due to a physical interaction between these two cell populations. As B‐1 cells are one of the main sources of interleukin (IL)‐10, we demonstrate in this study that adherent peritoneal cells from Xid mice produce significantly less amounts of this cytokine in culture when compared with IL‐10 production by cells from wild‐type mice. When B‐1 cells from IL‐10 knock‐out mice and macrophages from wild‐type mice were cocultured in transwell plates, the phagocytic index of macrophages was not altered demonstrating that B‐1 cells can influence the effector functions of macrophages in vitro via IL‐10 secretion.
Immunosuppression, interleukin‐10 synthesis and apoptosis are induced in rats inoculated with Cryptococcus neoformans glucuronoxylomannan
Authors: Chiapello Laura S. , Baronetti José L. , Aoki María P. , Gea Susana , Rubinstein Héctor , Masih Diana T. 
Published: October 2004

SummaryGlucuronoxylomannan (GXM) is the major Cryptococcus neoformans capsular polysaccharide and represents the main virulence factor of this fungus. In in vitro studies we have demonstrated previously that this acidic and high‐molecular‐weight polysaccharide suppresses lymphoproliferation, modulates cytokine production and promotes apoptosis in spleen mononuclear (Spm) cells from rats. In this study we demonstrate that these phenomena also occur in vivo after the intracardiac inoculation of GXM into normal Wistar rats. The results of this study show suppression of the proliferative response Spm cells to concanavalin A (Con A) or heat‐killed C. neoformans (HKCn) in the first 2 weeks after polysaccharide administration. In addition, increased levels of interleukin (IL)‐10 were produced by Con A‐stimulated Spm cells, coinciding with immunohistochemical GXM detection in the white pulp of spleen. In particular, high production of IL‐10 with diminution of IL‐2, interferon (IFN)‐γ and tumour necrosis factor (TNF)‐α synthesis were detected 14 days after GXM administration. In situ cell death detection by TdT‐mediated biotin–dUTP nick‐end labelling (TUNEL) reaction in sections of spleen, lung and liver demonstrates apoptosis in tissues with deposits of GXM. These data demonstrate the in vivo ability of GXM to modify cytokine synthesis by Spm cells and to promote host cell apoptosis.
Analysis of the mechanisms involved in the stimulation of neutrophil apoptosis by tumour necrosis factor‐α
Authors: Salamone Gabriela , Trevani Analía , Martínez Diego , Vermeulen Mónica , Gamberale Romina , Fernández‐Calotti Paula , Raiden Silvina , Giordano Mirta , Geffner Jorge 
Published: October 2004

SummaryWe have previously reported that human neutrophils pretreated with tumour necrosis factor‐α (TNF‐α) and then exposed to a variety of agents such as immune complexes, zymosan, phorbol 12‐myristate 13‐acetate (PMA), C5a, fMLP, or granulocyte–macrophage colony‐stimulating factor (GM‐CSF), undergo a dramatic stimulation of apoptosis, suggesting that TNF‐α is able to prime an apoptotic death programme which can be rapidly triggered by different stimuli. We report here that this response involves the participation of Mac‐1 (CD11b/CD18), is dependent on caspases 3, 8 and 9, and is associated with both a loss of mitochondrial transmembrane potential and a down‐regulation in expression of the anti‐apoptotic protein, Mcl‐1. Interestingly, we also found that the anti‐apoptotic cytokine interleukin‐1 (IL‐1) improves the ability of TNF‐α to promote apoptosis, supporting the notion than TNF‐α, acting together with IL‐1, may favour the depletion of neutrophils from the inflammatory areas during the course of acute inflammation.
CD27− CD4+ memory T cells define a differentiated memory population at both the functional and transcriptional levels
Authors: Schiött Åsa , Lindstedt Malin , Johansson‐Lindbom Bengt , Roggen Erwin , Borrebaeck Carl A. K. 
Published: October 2004

SummaryThe memory T‐cell population is a heterogeneous population, including both effector cells, which exert a direct secondary immune response, and resting or intermediate cells, which serve as a reservoir and exert a possible regulatory role. To further dissect the T‐cell memory population residing in the CD4+ CD45RO+ T‐cell pool, we studied the functional properties of memory populations identified by the CD27 marker. This marker clearly divides the memory population into two groups. One group consists of effector cells lacking CD27 and displaying a high antigen recall response. The other group consists of an intermediate memory population, displaying CD27. This latter group lacks an antigen recall response and requires costimulation for T‐cell receptor triggering. To evaluate the function of the CD27+ memory pool, we analysed the transcriptional profile, using high‐density microarray technology. These gene data strongly support the different functional profiles of CD27+ and CD27– memory populations, in terms of protein expression and the capacity to respond to antigen.
Interleukin‐10 suppression of myeloid cell activation — a continuing puzzle
Authors: Williams Lynn M. , Ricchetti Giuseppe , Sarma Usha , Smallie Timothy , Foxwell Brian M. J. 
Published: October 2004

SummaryEfforts to identify the signal transduction pathways used by interleukin‐10 (IL‐10) have resulted in limited success. The anti‐inflammatory effects elicited by IL‐10, and the mechanisms by which these are mediated, are still relatively unknown. Understanding the signalling mechanisms behind the suppression of cytokine expression by IL‐10 could be of potential therapeutic interest. Although the consensus is that the Janus kinase, Jak1, as well as the signal transducer and activator of transcription STAT3 are central, much controversy exists about the participation and roles of many other signalling pathways targeted by IL‐10. The mechanisms of cytokine suppression proposed by various groups have included transcriptional, post‐transcriptional and post‐translational regulation of IL‐10 target genes; nevertheless no unifying model has emerged thus far. Here we would like to highlight novel findings and discuss their implications in the context of current understanding of IL‐10 signalling.
Immunoreactive properties of peptide fractions of cow whey milk proteins after enzymatic hydrolysis
Authors: Wróblewska Barbara , Karamać Magdalena , Amarowicz Ryszard , Szymkiewicz Agata , Troszyńska Agnieszka , Kubicka Ewa 
Published: October 2004

SummaryCommercial whey protein concentrate (WPC) was hydrolysed with either Alcalase 2.4 FG (Novo Nordisk), or papain (Sigma) (in one‐step process) or with two enzymes (in two‐step process) to determine the changes in the immunoreactivity of α‐lactalbumin and β‐lactoglobulin. Enzymatic hydrolysis of WPC was performed by pH‐stat method. Hydrolysates were analysed using sodium dodecyl sulphate‐polyacrylamide gel electrophoresis, immunoblotting and size‐exclusion chromatography (SE‐HPLC). Immunoreactive properties of peptide fractions separated from the hydrolysates by fast protein liquid chromatography (FPLC) were determined using dot‐immunobinding and enzyme‐linked immunosorbent assay (ELISA) methods. Finally the sensory analysis was used to confirm organoleptic changes resulting from the application of different enzymes. The ‘two‐step’ process was observed to be the most effective however allergenic epitopes were still present, as it was found by ELISA with anti‐α‐la and anti‐β‐lg antibodies. The addition of papain as the second enzyme in the hydrolysis process contributed to the improvement of the sensory properties of WPC hydrolysate as compared with the Alcalase hydrolysate. Alcalase‐papain partially hydrolysated WPC can be found a promising base for production of the tolerogenic formula.
RIDASCREEN® FAST PEANUT, a rapid and safe tool to determine peanut contamination in food
Authors: Immer Ulrike , Reck Bernhard , Lindeke Stella , Koppelman Stef 
Published: October 2004

SummaryDuring the last years allergenic substances in food have gained more and more attention. Stricter rules for a peanut free food labelling demand a peanut contamination less than 5 ppm. The maintenance of a high quality of food products with regards to allergenic exposure is preferable warranted with rapid tests that can be utilized as an aid for suppliers to control raw materials, to avoid cross contamination during production and to check the final products. A rapid sandwich peanut ELISA was developed, which covers a measuring range of 2.5–20 ppm peanut (detection limit of 1.5 ppm). This test was successfully evaluated in an independent study under the supervision of the AOAC Research Institute, getting the AOAC certificate No. 030404. In‐house validation and independent validation data demonstrate the reliability, accuracy and precision of the test.
Fluorescent polarization immunoassay for sulphadiazine using a high specificity antibody
Authors: Murtazina Nailya R. , Eremin Sergei A. , Mozoleva Olga V. , Everest Sally J , Jim Brown A , Jackman Roy 
Published: October 2004

SummaryA rapid and sensitive fluorescent‐labelled polarization immunoassay for sulphadiazine (SDZ) is reported. Fluorescein‐labelled tracers were synthesized and a specific SDZ antibody has been used in the development of the method. The influence of the fluorescent label type on the immunoassay sensitivity was investigated. Homologous and non‐homologous tracer – antiserum combinations were varied to provide increased sensitivity of the immunoassay. A detection limit of 0.2 ng mL−1 for SDZ in 50 μL aqueous sample has been achieved. The cross‐reactivity of other sulphonamide drugs has also been examined.
Immunobiosensor determination of β‐agonists in urine using integrated immunofiltration clean‐up
Authors: Annette Johansson Marie , Hellenäs Karl‐Erik 
Published: October 2004

SummaryA fast immunobiosensor assay for screening of β‐agonists in urine, using integrated immunofiltration clean‐up with 30 kDa cutoff centrifugal filter, was developed. The same antibody aliquot was used for clean‐up and biosensor analysis, and no organic solvent was needed. Matrix interference from urine was efficiently reduced. Two antibodies with different cross‐reactivities were compared. Although both antibodies were able to detect clenbuterol (CBL) below 1 ng mL−1, a polyclonal antibody directed against salbutamol was chosen because of its ability to recover several β‐agonists. The decision limit (CCα) of the final assay was 0.29 ng mL−1, and an α‐error of 1% gave a β‐error of <1% for twenty reference bovine samples spiked with 1 ng mL−1 CBL.
High levels of p53 protein expression do not correlate with p53 mutations in hepatocellular carcinoma
Authors: Anzola M. , Saiz A. , Cuevas N. , Lopez‐Martinez M. , Martinez de Pancorbo M. A. , Burgos J. J. 
Published: October 2004

Summary.  To determine the relationship between p53 altered expression and p53 mutations in hepatocellular carcinoma (HCC), we analysed p53 protein immunohistochemically and assessed the presence of mutations in exons 4–8 of the p53 gene using SSCP assay in 117 HCCs corresponding to 78 patients. We also determined the relationship of p53 expression with cellular proliferation by immunostaining with monoclonal antibodies to Ki‐67. We found significant levels of p53 protein expression in 23.1% of the 117 cases studied, but identified mutations in only 12 cases (10.3%). Only four of the p53‐positive cases had mutations in the regions analysed. Six of the cases that displayed mutations at p53 gene were negative for immunohistochemical analysis (IHC) and two cases showed positive immunoreactivity in the cytoplasm of the cell.In conclusion, strong IHC reactivity for p53 protein is not an indicator of the presence of p53 gene mutations at exons 4–8 in HCC. Thus, p53 loss of function in HCC should be evaluated both by p53 mutation analysis and p53 protein expression, as both give complementary information about p53 status.