Leukotrienes Play a Role in the Control of Parasite Burden in Murine Strongyloidiasis Eleuza R. Machado, Marlene T. Ueta, Elaine V. Lourenço, Fernanda F. Anibal, Carlos Artério Sorgi, Edson G. Soares, This information is current as Maria C. Roque-Barreira, Alexandra I. Medeiros and Lúcia of October 6, 2021. H. Faccioli J Immunol 2005; 175:3892-3899; ; doi: 10.4049/jimmunol.175.6.3892 http://www.jimmunol.org/content/175/6/3892 Downloaded from References This article cites 53 articles, 12 of which you can access for free at: http://www.jimmunol.org/content/175/6/3892.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 6, 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 Leukotrienes Play a Role in the Control of Parasite Burden in Murine Strongyloidiasis1 Eleuza R. Machado,* Marlene T. Ueta,† Elaine V. Lourenc¸o,‡ Fernanda F. Anibal,* Carlos Arte´rio Sorgi,* Edson G. Soares,§ Maria C. Roque-Barreira,‡ Alexandra I. Medeiros,* and Lu´cia H. Faccioli2* It is clear that leukotrienes mediate inflammatory response; new aspects of leukotriene function have recently been described. In this study, we demonstrate that leukotrienes are key chemical mediators in the control of parasite burdens in mice infected with Strongyloides venezuelensis. High leukotriene levels were detected in the lungs and small intestines of Swiss mice. In infected Swiss mice treated with MK886, a leukotriene synthesis inhibitor, numbers of adult worms, and eggs/g/feces were greater than in infected-only animals. The MK886 treatment inhibited leukotriene B4 production in the lungs and small intestines, albeit on ؊/؊ different postinfection days. Similarly, parasite burdens and eggs/g/feces were greater in 5-lipoxygenase mice than in wild-type Downloaded from animals. These observation were confirmed by histopathological study of the duodena. We subsequently observed significant lower numbers of eosinophils and mononuclear cells in the blood, peritoneal cavity fluid, and bronchoalveolar lavage fluid of Swiss mice treated with MK886. In the lung parenchyma of infected animals, MK886 significantly inhibited synthesis of IL-5 at the beginning of infection, whereas levels of IL-12 increased progressively throughout the postinfection period. However, levels of leukotriene C4, ␣ ␥ PGE2, TNF- , IL-3, IL-4, IFN- , and IL-10 were comparable between the treated and untreated groups. Nevertheless, IgE and IgG1 (but not IgG2a) synthesis was also significantly inhibited by MK886 administration. Therefore, in S. venezuelensis-infected http://www.jimmunol.org/ mice, adult worm and egg burdens are leukotriene dependent. These findings indicate potential immunostimulatory strategies involving leukotriene administration, and may serve as an alert to physicians treating Strongyloides stercoralis-infected patients presenting asthma-like symptoms because use of 5-lipoxygenase inhibitors may worsen the infection. The Journal of Immunology, 2005, 175: 3892–3899. he incidence of strongyloidiasis has increased dramati- ten results in the death of the host (6, 7). To date, the cause of this cally, mainly in patients presenting altered immune status intense proliferation is unknown. accompanied by malnutrition, having undergone organ or In human hosts and in murine models, the immune response to T by guest on October 6, 2021 bone marrow transplantation, with acquired immunodeficiency Strongyloides sp is characterized by intraepithelial and tissue in- syndrome, or receiving cancer chemotherapy (1–5). Many patients crease of eosinophils (8, 9), as well as by intestinal mastocytosis chronically infected with Strongyloides stercoralis are asymptom- (10, 11) and production of Th2-type cytokines such as IL-3, IL-4, atic, whereas others present a variety of symptoms, most related to and IL-5 (11–13). Production of IgA, IgE, IgG, and IgM Abs has the skin or gastrointestinal system, and many S. stercoralis infec- also been demonstrated (14, 15). However, far less attention has tions evolve to a cure (6). When the host-parasite balance is dis- been given to the role of leukotrienes in host defense during turbed, as occurs in immunocompromised patients, intestinal strongyloidiasis infection, in which they could act as mediators of worms may reproduce excessively and invade organs outside their antiparasitic activity. Leukotrienes are generated via the 5-lipoxy- normal migratory pathways. Such dissemination, if untreated, of- genase (5-LO)3 pathway in the arachidonic acid metabolism and have been implicated in many inflammatory and allergic diseases (16–19). Various authors have described new leukotriene immune response functions (20–23) and antimicrobial activity (24, 25). *Departamento de Ana´lises Clı´nicas, Toxicolo´gicas e Bromatolo´gicas, Faculdade de Cieˆncias Farmaceˆuticas de Ribeira˜o Preto, Universidade de Sa˜o Paulo, Ribeira˜o Preto, More recently, we demonstrated that leukotrienes are essential to Sa˜o Paulo, Brazil; †Departamento de Parasitologia, Instituto de Biologia, Univer- efficient pulmonary antifungal host defense because they modulate sidade Estadual de Campinas, Sa˜o Paulo, Brazil; ‡Departamento de Biologia Celular, Molecular e Bioagentes Patogeˆnicos, Faculdade de Medicina de Ribeira˜o Preto, Uni- NO and cytokine production during infection (26). The importance versidade de Sa˜o Paulo, Ribeira˜o Preto, Sa˜o Paulo, Brazil; and §Departamento de of leukotrienes in controlling helminth parasite burdens is still un- Patologia, Faculdade de Medicina de Ribeira˜o Preto, Universidade de Sa˜o Paulo, known, and very little information is available on the subject. It Ribeira˜o Preto, Sa˜o Paulo, Brazil has been shown that levels of leukotriene B (LTB ) and leuko- Received for publication November 9, 2004. Accepted for publication June 30, 2005. 4 4 triene C (LTC ) are elevated in the intestinal mucosa of lambs The costs of publication of this article were defrayed in part by the payment of page 4 4 charges. This article must therefore be hereby marked advertisement in accordance infected with Trichostrongylus colubriformis (27–29). The authors with 18 U.S.C. Section 1734 solely to indicate this fact. suggested that these lipid mediators are associated with larval re- 1 This study was supported by grants from the Fundac¸a˜o de Amparo a`Pesquisa do jection and exclusion. Estado de Sa˜o Paulo (02/12856-2), the Conselho Nacional de Desenvolvimento Ci- entı´fico e Tecnolo´gico, and the Coordenac¸a˜o de Aperfeic¸oamento de Pessoal de Ensino Superior, Brazil. 2 Address correspondence and reprint requests to Dr. Lu´cia H. Faccioli, Departamento de Ana´lises Clı´nicas, Toxicolo´gicas e Bromatolo´gicas, Faculdade de Cieˆncias Farma- 3 Abbreviations used in this paper: 5-LO, 5-lipoxygenase; BALF, bronchoalveolar ceˆuticas de Ribeira˜o Preto, Universidade de Sa˜o Paulo - Avenida do Cafe´, s/n. Ri- lavage fluid; LTB4, leukotriene B4; LTC4, leukotriene C4; PCF, peritoneal cavity beira˜o Preto, Sa˜o Paulo, Brasil, 14.040-903. E-mail address: [email protected] fluid; WT, wild type. Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 The Journal of Immunology 3893 In the present study, we investigated the effects of inhibition or 1500 ϫ g, filtered, and stored at Ϫ70°C until assay. Eicosanoids were absence of leukotrienes during Strongyloides venezuelensis infec- quantified using commercial ELISA kits obtained from Amersham Bio- tion in mice. Our results reveal for the first time that leukotrienes sciences. Commercially available ELISA Abs were used to measure TNF-␣, IL-3, IL-4, IL-5, IL-10, IL-12, and IFN-␥, according to manufac- play an essential role in controlling parasite burdens, as well as in turer instructions (BD Pharmingen). Sensitivities were Ͼ10 pg/ml. altering the parasite reproductive cycle and eliminating the para- sites themselves. Measurement of Abs in sera Specific IgE, IgG1, and IgG2a were determined in mice sera by ELISA, Materials and Methods according to manufacturer instructions (Technical Data Sheet; BD Pharm- Animals ingen). The plates were coated with S. venezuelensis alkaline extract at a concentration of 20 ␮g/ml in carbonate buffer at pH 9.6 for 18 h at 4°C. Male Swiss mice weighing 16–25 g and male Wistar rats weighing 120– After three washes in PBS containing 0.05% Tween 20 (pH 7.4), nonspe- 180 g were obtained from the animal facilities of the Central de Bioterismo cific binding sites were blocked with a buffer composed of the same PBS- da Universidade Estadual de Campinas and Faculdade de Cieˆncias Farma- Tween 20 solution plus 1% BSA for1hat37°C. Serum samples (50 ␮l ceˆuticas de Ribeira˜o Preto, Universidade de Sa˜o Paulo. Male mice lacking Ϫ Ϫ each) were added to wells at a dilution of 1/20 (for measurement of IgG1 the 5-LO enzyme gene (5-LO / ) and weighing 18–25 g were obtained and IgG2a) or 1/5 (for measurement of IgE), both in blocking buffer. To from The Jackson Laboratory, and age-matched male wild-type (WT) mice measure IgG1 and IgG2a, plates were then incubated at room temperature (background, strain 129) were used as controls. All experiments were ap- for 1 h. For IgE measurement, however, the plate was incubated at 4°C for proved by and conducted in accordance with guidelines established by the 24 h and washed five times.