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Selective Inhibition of IgG-Mediated Phagocytosis in Gelsolin-Deficient Murine Neutrophils This information is current as Lena Serrander, Petra Skarman, Birgitta Rasmussen, Walter of October 2, 2021. Witke, Daniel P. Lew, Karl-Heinz Krause, Olle Stendahl and Oliver Nüße J Immunol 2000; 165:2451-2457; ; doi: 10.4049/jimmunol.165.5.2451 http://www.jimmunol.org/content/165/5/2451 Downloaded from References This article cites 46 articles, 26 of which you can access for free at: http://www.jimmunol.org/content/165/5/2451.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 2, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2000 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. Selective Inhibition of IgG-Mediated Phagocytosis in Gelsolin-Deficient Murine Neutrophils1 Lena Serrander,2* Petra Skarman,† Birgitta Rasmussen,† Walter Witke,‡ Daniel P. Lew,* Karl-Heinz Krause,3* Olle Stendahl,† and Oliver Nu¨ße4* Phagocytosis and the microbicidal functions of neutrophils require dynamic changes of the actin cytoskeleton. We have investi- gated the role of gelsolin, a calcium-dependent actin severing and capping protein, in peripheral blood neutrophils from gelsolin- null (Gsn؊) mice. The phagocytosis of complement opsonized yeast was only minimally affected. In contrast, phagocytosis of IgG-opsonized yeast was reduced close to background level in Gsn؊ neutrophils. Thus, gelsolin is essential for efficient IgG- but not complement-mediated phagocytosis. Furthermore, attachment of IgG-opsonized yeast to Gsn؊ neutrophils was reduced (ϳ50%) but not to the same extent as ingestion (ϳ73%). This was not due to reduced surface expression of the Fc␥-receptor or its lateral mobility. This suggests that attachment and ingestion of IgG-opsonized yeast by murine neutrophils are actin-dependent Downloaded from and gelsolin is important for both steps in phagocytosis. We also investigated granule exocytosis and several steps in phagosome processing, namely the formation of actin around the phagosome, translocation of granules, and activation of the NADPH-oxidase. All these functions were normal in Gsn؊ neutrophils. Thus, the role of gelsolin is specific for IgG-mediated phagocytosis. Our data suggest that gelsolin is part of the molecular machinery that distinguishes complement and IgG-mediated phagocytosis. The latter requires a more dynamic reorganization of the cytoskeleton. The Journal of Immunology, 2000, 165: 2451–2457. olymorphonuclear leukocytes (neutrophils) constantly re- olites and granular enzymes into the phagosomal space, demand http://www.jimmunol.org/ arrange their actin cytoskeleton to perform chemotaxis and rapid and dynamic cytoskeletal rearrangements. Gelsolin consti- phagocytosis. Such rearrangements depend on actin sev- tutes about 1% of the total neutrophil protein and is a good can- P 2ϩ ering and capping proteins of which gelsolin is the best-character- didate to mediate Ca -dependent actin rearrangements (6). ized example (1). Gelsolin is an 82-kDa cytoplasmic protein Phagocytosis in neutrophils is well studied, but its molecular ϩ widely expressed in mammalian cells. Upon activation by Ca2 , mechanisms are still poorly understood. The two main pathways gelsolin severs actin filaments, binds, and caps barbed ends of are complement-receptor (CR)-mediated and Fc␥-receptor actin filaments. Gelsolin-actin complexes dissociate in the pres- (Fc␥R)-mediated phagocytosis (reviewed in Ref. 7). The important Ϫ 5 ence of polyphosphoinositides (2, 3). Gelsolin-null (Gsn ) mice receptors are CR1, CR3, and CR4 for CR-mediated phagocytosis by guest on October 2, 2021 Ϫ have normal embryonic development and longevity (4). Gsn and Fc␥R II and Fc␥R III for Fc␥R-mediated phagocytosis in non- mice show a prolonged bleeding time due to decreased platelet stimulated neutrophils. shape changes. Migration of neutrophils into peritoneal exudates is Binding and clustering of CRs by complement-opsonized par- delayed, and fibroblasts have excessive stress fibers and decreased Ϫ ticles mediates serine phosphorylation of the receptor and the for- motility. The GTP-binding protein rac is overexpressed in Gsn mation of punctuate structures containing F-actin, paxillin, ␣-ac- tissues and regulates gelsolin in fibroblasts (5). tinin, and vinculin assembled to the membrane (8). This leads to a Neutrophil functions, such as adhesion, chemotaxis, phagocy- sinking of the opsonized particle into the membrane without pro- tosis and finally microbial killing by secretion of oxidative metab- truding pseudopods (9). Fc␥R-mediated phagocytosis has been shown to have another morphology with protruding pseudopodia *Divison of Infectious Diseases, University of Geneva, Switzerland; †Department for encircling the prey, thus demanding another kind of actin remod- Medical Microbiology, University of Linko¨ping, Sweden; and ‡European Molecular eling (9). Fc␥R-receptor stimulation mediates signaling via im- Biology Laboratory Mouse Biology Program, Monterotondo, Italy mune receptor tyrosine-based activation motifs (ITAM) with the Received for publication August 16, 1999. Accepted for publication June 16, 2000. initiation of a signaling cascade involving phosphorylation of syk, The costs of publication of this article were defrayed in part by the payment of page src-kinases activation of PI3 kinases, rac and phospholipase C, that charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. stimulate protein kinase C-activation and calcium release from in- 1 This work was supported by Grants from the Swiss National Fund 31-45891.95 and tracellular stores (7–9). 31-50786.97, Swedish Medical Council no 5968, the Swedish Rheumatoid Founda- An actin ring that slowly dissolves after ingestion is completed tion, the Foundation Carlos et Elsie de Reuter, and the Socie´te´ Acade´mique de encircles the phagosome (10). The actin ring may be a barrier, Gene`ve. preventing granules from fusion with the phagosome. Gelsolin 2 Address correspondence and reprint requests to Dr. Lena Serrander, Division of Infectious Diseases, University Hospital of Geneva, CH-1211 Geneva 14, Switzer- translocates to the actin ring and might be important for its re- land. E-mail address: [email protected] moval (P.S. and O.S., unpublished observations). Intracellular 2ϩ 3 Current address: Laboratory Biology of Aging, Department of Geriatrics, University Ca stores and granules also translocate to the phagosome and Hospitals of Geneva, Chemin Petit-Bel-Air 2, CH-1225 Cheˆne-Bourg, Switzerland. phagolysosome fusion takes place (11). In neutrophils, this is a 4 Current address: Immunology Laboratory, Science Faculty, University Nancy 1, calcium-dependent process (12). B. P. 239, F-54506 Vandoeuvre-les-Nancy, France. Extracellular secretion of granule proteins is another neutrophil 5 Abbreviations used in this paper: GsnϪ, Gelsolin-null; E.S., external solution; Fc␥R, Fc␥ receptor; CR, complement receptor; MPO, myeloperoxidase; NBT, nitro- activity, important for killing, but also causing tissue damage. The blue tetrazolium; FRAP, fluorescence recovery after photobleaching. cortical actin network may form a physical barrier that prevents Copyright © 2000 by The American Association of Immunologists 0022-1767/00/$02.00 2452 THE ROLE OF GELSOLIN IN NEUTROPHIL PHAGOCYTOSIS granules from reaching the plasma/phagosomal membrane. The FACS analysis role of gelsolin could be to break up this barrier. It has indeed been Mouse neutrophils (20,000 cells/tube) were pelleted and resuspended in 0.1 shown that actin depolymerization in itself is sufficient to induce ml cold 4% paraformaldehyde. After 30 min, the cells were washed twice secretion in some cell types and that gelsolin can stimulate secre- and incubated for 1 h with 0.1 ml either rat monoclonal anti-mouse tion (13–15). CD16/32 (clone 2.4G2) or anti-mouse CD18 (clone C71/16) diluted 1/100 Ϫ in 0.1% BSA/PBS. After one wash, 0.1 ml of an FITC-labeled secondary We have investigated neutrophils from Gsn mice to study the Ј Ab, goat anti-rat F(ab )2, -fragments diluted 1/200 in 0.1% BSA/PBS sup- role of gelsolin in neutrophil functions. Our results show the fea- plemented with 5% goat IgG, was added. Cells were washed and resus- sibility of using transgenic mice to study the specific role of a pended in 0.2 ml PBS. protein in peripheral blood neutrophils. Gelsolin is the prototype of Cell labeling and fluorescence recovery after photobleaching actin severing and capping proteins. It is thought to play a central (FRAP) role in the actin dynamics of phagocytes. We show here that gel- solin plays an important role in Fc␥R-mediated phagocytosis, but Neutrophils (106/ml) were incubated at 4°C for 1 h with 100 ␮l Krebs- not in several other phagocyte functions.
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  • Neuronal Calcium Sensor-1 Enhancement of Insp3 Receptor Activity Is Inhibited by Therapeutic Levels of Lithium

    Neuronal Calcium Sensor-1 Enhancement of Insp3 Receptor Activity Is Inhibited by Therapeutic Levels of Lithium

    Neuronal calcium sensor-1 enhancement of InsP3 receptor activity is inhibited by therapeutic levels of lithium Christina Schlecker, … , Klara Szigeti-Buck, Barbara E. Ehrlich J Clin Invest. 2006;116(6):1668-1674. https://doi.org/10.1172/JCI22466. Research Article Neuroscience Regulation and dysregulation of intracellular calcium (Ca2+) signaling via the inositol 1,4,5-trisphosphate receptor (InsP3R) has been linked to many cellular processes and pathological conditions. In the present study, addition of neuronal calcium sensor-1 (NCS-1), a high-affinity, low-capacity, calcium-binding protein, to purified InsP3R type 1 (InsP3R1) increased the channel activity in both a calcium-dependent and -independent manner. In intact cells, enhanced expression of NCS-1 resulted in increased intracellular calcium release upon stimulation of the phosphoinositide signaling pathway. To determine whether InsP3R1/NCS-1 interaction could be functionally relevant in bipolar disorders, conditions in which NCS-1 is highly expressed, we tested the effect of lithium, a salt widely used for treatment of bipolar disorders. Lithium inhibited the enhancing effect of NCS-1 on InsP3R1 function, suggesting that InsP3R1/NCS-1 interaction is an essential component of the pathomechanism of bipolar disorder. Find the latest version: https://jci.me/22466/pdf Research article Neuronal calcium sensor-1 enhancement of InsP3 receptor activity is inhibited by therapeutic levels of lithium Christina Schlecker,1,2,3 Wolfgang Boehmerle,1,4 Andreas Jeromin,5 Brenda DeGray,1 Anurag Varshney,1 Yogendra Sharma,6 Klara Szigeti-Buck,7 and Barbara E. Ehrlich1,3 1Department of Pharmacology, Yale University, New Haven, Connecticut, USA. 2Department of Neuroscience, University of Magdeburg, Magdeburg, Germany.