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Hemozoin Induces Macrophage Chemokine Expression Through Oxidative Stress-Dependent and -Independent Mechanisms This Information Is Current As of October 7, 2021

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Hemozoin Induces Macrophage Chemokine Expression Through Oxidative Stress-Dependent and -Independent Mechanisms This Information Is Current As of October 7, 2021 Hemozoin Induces Macrophage Chemokine Expression through Oxidative Stress-Dependent and -Independent Mechanisms This information is current as of October 7, 2021. Maritza Jaramillo, Marianne Godbout and Martin Olivier J Immunol 2005; 174:475-484; ; doi: 10.4049/jimmunol.174.1.475 http://www.jimmunol.org/content/174/1/475 Downloaded from References This article cites 54 articles, 24 of which you can access for free at: http://www.jimmunol.org/content/174/1/475.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 7, 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 © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology Hemozoin Induces Macrophage Chemokine Expression through Oxidative Stress-Dependent and -Independent Mechanisms1 Maritza Jaramillo,† Marianne Godbout,* and Martin Olivier2†* Chemokine production has been associated with the immunopathology related to malaria. Previous findings indicated that he- mozoin (HZ), a parasite metabolite released during schizogeny, might be an important source of these proinflammatory mediators. In this study we investigated the molecular mechanisms underlying HZ-inducible macrophage (M␾) chemokine mRNA expres- sion. We found that both Plasmodium falciparum HZ and synthetic HZ increase mRNA levels of various chemokine transcripts (MIP-1␣/CCL3, MIP-1␤/CCL4, MIP-2/CXCL2, and MCP-1/CCL2) in murine B10R M␾. The cellular response to HZ involved ERK1/2 phosphorylation, NF-␬B activation, reactive oxygen species (ROS) generation, and ROS-dependent protein-tyrosine phosphatase down-regulation. Selective inhibition of either I␬B␣ or the ERK1/2 pathway abolished both NF-␬B activation and Downloaded from chemokine up-regulation. Similarly, blockage of HZ-inducible M␾ ROS with superoxide dismutase suppressed chemokine in- duction, strongly reduced NF-␬B activation, and restored HZ-mediated M␾ protein-tyrosine phosphatase inactivation. In con- trast, superoxide dismutase had no effect on EKR1/2 phosphorylation by HZ. Collectively, these data indicate that HZ triggers ROS-dependent and -independent signals, leading to increased chemokine mRNA expression in M␾. Overall, our findings may help to better understand the molecular mechanisms through which parasite components, such as HZ, modulate the immune response during malaria infection. The Journal of Immunology, 2005, 174: 475–484. http://www.jimmunol.org/ lasmodium falciparum (Pf),3 the etiological agent of the MIP-1␣/CCL3 and IL-8/CXCL8 were detected during acute infec- most lethal form of human malaria, is responsible for tion (4). In placental malaria, increased MIP-1␣, IL-8, and MCP- P 300–500 million clinical cases and 1–3 million deaths 1/CCL2 mRNA expression was associated with placental mono- annually (1). Accumulating evidence indicates that in addition to cyte recruitment (5), a risk factor for low birth weight (6). In parasite virulence factors, the host’s immunological response (e.g., addition, experimental models of cerebral malaria have underlined proinflammatory cytokines and chemokines) also participates in the importance of the neutrophil (N␾) population in the develop- malaria pathophysiology, including tissue cell damage, microvas- ment of this pathology (7) and have shown the ability of both N␾ cular obstruction, severe anemia, and cerebral malaria (2). Because and macrophages (M␾) to express elevated chemokine mRNA lev- by guest on October 7, 2021 of their ability to induce activation and recruitment of specific els (8). leukocyte subsets to the sites of infection, chemokines influence In an attempt to identify the parasite components that are re- the immune system, thereby further inducing inflammatory condi- sponsible for the activation of the host immune response during tions (3). In Pf malaria-infected patients, elevated serum levels of malaria, we (9, 10) and others (11–13) have focused on evaluating the proinflammatory properties of hemozoin (HZ) or malarial pig- ment. HZ crystals result from aggregated heme, which is produced *Research Institute of McGill University Health Center, Center for the Study of Host Resistance, Departments of Medicine, Microbiology, and Immunology, McGill Uni- during hemoglobin digestion inside the host RBC (14). This pig- versity, Montre´al, Que´bec, Canada; and †Centre de Recherche en Infectiologie, Cen- ment is released after schizogeny, along with merozoites, as the tre Hospitalier Universitaire de Que´bec, Pavillon Centre Hospitalier de l’Universite´ ␾ Laval, and De´partement de Biologie Me´dicale, Faculte´deMe´decine, Universite´La- RBC bursts and is phagocytosed by circulating monocytes, N , val, Ste-Foy, Que´bec, Canada and resident M␾ (reviewed by Arese and Schwarzer (15)). Both Pf Received for publication February 6, 2004. Accepted for publication October HZ and synthetic HZ (sHZ), which is structurally identical with the 22, 2004. native pigment (16), induced the release of MIP-1␣ and MIP-1␤/ The costs of publication of this article were defrayed in part by the payment of page CCL4 as well as of TNF-␣ and IL-1␤ in murine M␾ and human charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact. PBMC (11, 12). These observations were also supported by our in vivo data indicating that sHZ leads to liver chemokine and cyto- 1 This work was supported by grants from the Canadian Institutes in Health Research (CIHR; to M.O.). M.O. is member of a CIHR Group in Host-Pathogen Interactions, kine mRNA up-regulation in i.v. injected mice and to significant a recipient of a CIHR Investigator Award, and a Burroughs Wellcome Fund Awardee leukocyte recruitment and proinflammatory mediator release in the in Molecular Parasitology. M.J. is a recipient of a Ministe`redel’E´ ducation du Que´bec Ph.D. studentship. M.G. is a recipient of a Fonds pour la Formation de Chercheurs et air pouch lumen of mice inoculated intradermally (10). In line with l’Aide a`la Recherche M.Sc. studentship. these findings, Biswas and colleagues (13) reported the presence of 2 Address correspondence and reprint requests to Dr. Martin Olivier, Department of Abs against Pf HZ among complicated malaria patients, which had Microbiology and Immunology, McGill University, 3775 University Street, Duff inhibitory effects on HZ-dependent monocyte cytokine production. Medical Building (Room 600), Montre´al, Que´bec, Canada H3A 2B4. E-mail address: [email protected] Although these studies strongly suggest that HZ is likely to play a key role in proinflammatory mediator overproduction, the mech- 3 Abbreviations used in this paper: Pf, Plasmodium falciparum; HZ, hemozoin; IKK, I␬B kinase; M␾, macrophage; N␾, neutrophil; PTK, protein-tyrosine kinase; PTP, anism by which such regulation occurs remains unexplored. In the . protein-tyrosine phosphatase; O2, superoxide anion; Poly B, polymyxin B; pNPP, present study we demonstrate that both Pf HZ and sHZ increase para-nitrophenyl phosphate; redox, reduction-oxidation; RPA, RNase protection as- ␣ ␤ say; ROS, reactive oxygen species; sHZ, synthetic hemozoin or ␤-hematin; SOD, mRNA levels of various chemokines (MIP-1 , MIP-1 , MIP-2, superoxide dismutase. and MCP-1) in murine M␾ through a mechanism that involves Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 476 HZ-INDUCIBLE M␾ CHEMOKINE MODULATION activation of oxidative stress-dependent and -independent path- study (ϳ26–29 nmol/ml heme content) could be easily released by a pa- ways. A better understanding of the transductional signals through tient with a mild parasitemia load. which HZ modulates the host immune response might be helpful in RNase protection assay (RPA) defining specific therapeutic targets to tame proinflammatory me- diator overproduction during malaria infection. mRNA expression studies were performed using an RPA kit (RiboQuant; BD Pharmingen), as we described previously (22). Total RNA was isolated from stimulated cells with TRIzol reagent (Invitrogen Life Technologies) Materials and Methods according to the manufacturer’s protocol. One of the multiprobe templates Materials was labeled with [␣-32P]dUTP using T7 RNA polymerase. Then, 3 ϫ 105 cpm of labeled probe was allowed to hybridize with 10 ␮g of total RNA for Hemin chloride, polymyxin B (Poly B) sulfate, LPS (Escherichia coli, 16 h at 56°C. mRNA probe hybrids were treated with RNase A and phenol- serotype 0111:B4), 3% H2O2 (v/v), and superoxide dismutase (SOD) were chloroform-extracted. Protected hybrids were resolved on a 5% denaturing purchased from Sigma-Aldrich. Isotopes [␣-32P]dUTP (3000 Ci/mmol) polyacrylamide sequencing gel and exposed to radiographic film overnight and [␥-32P]dATP (3000 Ci/mmol) were obtained from PerkinElmer. BAY at Ϫ80°C. Laser densitometry was performed using an ␣Imager 2000 dig- 11-7082 was purchased from BIOMOL. Apigenin and PD 98059 were ital imaging and analysis system (␣Innotech). The multiprobe templates obtained from Calbiochem. (BD Pharmingen) used in this study were mCK-5, for the murine chemo- kines lymphotactin, RANTES, eotaxin, MIP-1␣, MIP-1␤, MIP-2, inducing Cell and culture conditions protein-10, MCP-1, and TCA-3, and a custom template for murine cyto- ␣ ␤ ␥ ␾ kines IL-4, IL-12p40, IL-10, IL-1 , IL-1 , IL-2, IL-6, and IFN- . The The murine M cell line B10R, derived from the bone marrow of template sets included housekeeping genes ml-32 and/or GAPDH. B10A.Bcgr (B10R) mice (17), was provided by Dr.
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