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      Cytokine Release Pathway in Mononuclear Cells Stimulated in vitro by Dialysis Membranes

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          Abstract

          Background: Among the possible variables associated with cytokine activation in hemodialysis, filter membrane has been reported to be a major factor and monocytes adherent to the membrane have been suggested as a possible source for cytokine synthesis. Methods: In order to exclude variables other than the direct cell-to-membrane interaction, suspensions of peripheral blood monocytes isolated from donors’ blood were incubated for 180 min at 37°C in Petri dishes coated with cuprophan (Cu) or polyacrylonitrile (AN69S) or polysulfone (PS). Total RNA was purified, reverse transcribed in cDNA and amplified by polymerase chain reaction (PCR) primed with specific oligomers for determining IL-1β and TNF-α gene expression. For Western blot analysis, cell homogenates and supernatants were electrophoresed and transferred to a polyvinylidene difluoride membrane and the membrane was then incubated with polyclonal antibodies specific for the detection of IL-1β and TNF-α. Results: Semi-quantitation of targets for PCR (using the technique of limiting dilutions and referring to actin and glyceraldehyde-3-phosphate dehydrogenase as noninducible molecule) revealed a comparable increase at 180 min (p < 0.05) in IL-1β and TNF-α mRNA in monocytes incubated with polyacrylonitrile, PS and Cu membranes. In 2 of 10 experiments pro-IL-1β was detected in monocytes interacting with Cu, PS and AN69S membrane. However, the extent of extracellular release of mature protein was greater for Cu. Conclusion: Some dissociation between transcriptional and translational events was detected in experiments using donors’ blood in vitro. More specifically, synthetic membranes (both polyacrylonitrile and PS) were found to be as active as Cu in inducing IL-1β and TNF-α mRNA expression, but less effective in promoting the extracellular release of proinflammatory cytokine products.

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          Most cited references 5

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          A novel heterodimeric cysteine protease is required for interleukin-1 beta processing in monocytes.

          Interleukin-1 beta (IL-1 beta)-converting enzyme cleaves the IL-1 beta precursor to mature IL-1 beta, an important mediator of inflammation. The identification of the enzyme as a unique cysteine protease and the design of potent peptide aldehyde inhibitors are described. Purification and cloning of the complementary DNA indicates that IL-1 beta-converting enzyme is composed of two nonidentical subunits that are derived from a single proenzyme, possibly by autoproteolysis. Selective inhibition of the enzyme in human blood monocytes blocks production of mature IL-1 beta, indicating that it is a potential therapeutic target.
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            Review: transcriptional and post-transcriptional regulation of interleukin 1 gene expression.

            Interleukin 1 alpha (IL-1 alpha) and beta (IL-1 beta) are proinflammatory cytokines that are encoded by distinct genes, but share most biological activities. During the past several years, intense investigation has focused on elucidating the molecular basis for the regulation of IL-1 alpha and beta gene expression. While the overall organization of both genes is conserved in mammals, the DNA sequence homology is surprisingly limited. This supports the growing body of evidence suggesting that each gene is regulated by distinct cis- and transacting elements. Most recently, novel regulatory DNA sequence elements and several nuclear regulatory proteins have been identified, which ultimately participate in the control of IL-1 beta gene transcription. In addition to transcriptional controls, the stability of IL-1 mRNA can be selectively regulated by various inducing stimuli and other cytokines. Taken together, these transcriptional and post-transcriptional regulatory mechanisms provide stringent, yet flexible, control over expression of the IL-1 alpha and beta genes.
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              Human Monocyte Interleukin-1β Posttranslational Processing

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                Author and article information

                Journal
                AJN
                Am J Nephrol
                10.1159/issn.0250-8095
                American Journal of Nephrology
                S. Karger AG
                0250-8095
                1421-9670
                2002
                December 2002
                07 October 2002
                : 22
                : 5-6
                : 509-514
                Affiliations
                aCattedre e Scuola di Specializzazione di Patologia Clinica e Dipartimento di Medicina e Oncologia Sperimentale, Università di Torino; bCentro di Ricerche di Immunopatologia e Documentazione su Malattie Rare (CMID), Presidi Ospedalieri ASL4, sede Luigi Einaudi, Torino, Italia
                Article
                65288 Am J Nephrol 2002;22:509–514
                10.1159/000065288
                12381952
                © 2002 S. Karger AG, Basel

                Copyright: All rights reserved. No part of this publication may be translated into other languages, reproduced or utilized in any form or by any means, electronic or mechanical, including photocopying, recording, microcopying, or by any information storage and retrieval system, without permission in writing from the publisher. Drug Dosage: The authors and the publisher have exerted every effort to ensure that drug selection and dosage set forth in this text are in accord with current recommendations and practice at the time of publication. However, in view of ongoing research, changes in government regulations, and the constant flow of information relating to drug therapy and drug reactions, the reader is urged to check the package insert for each drug for any changes in indications and dosage and for added warnings and precautions. This is particularly important when the recommended agent is a new and/or infrequently employed drug. Disclaimer: The statements, opinions and data contained in this publication are solely those of the individual authors and contributors and not of the publishers and the editor(s). The appearance of advertisements or/and product references in the publication is not a warranty, endorsement, or approval of the products or services advertised or of their effectiveness, quality or safety. The publisher and the editor(s) disclaim responsibility for any injury to persons or property resulting from any ideas, methods, instructions or products referred to in the content or advertisements.

                Page count
                Figures: 4, References: 24, Pages: 6
                Product
                Self URI (application/pdf): https://www.karger.com/Article/Pdf/65288
                Categories
                Clinical Study

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