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Qualitative and quantitative immunohistochemical evaluation of iNOS expression in the spleen of dogs naturally infected with Leishmania chagasi

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Abstract

Nitric oxide (NO), the product of the nitric oxide synthase enzymes has been detected in Leishmania-infected animals. Besides its role on the immunity to infection, the role of NO and the inducible nitric oxide synthase (iNOS) in the pathogenesis of canine visceral leishmaniasis (CVL) is not well understood. This study aimed at evaluating immunohistochemically the iNOS expression in the spleen of dogs naturally infected (ID) with Leishmania (L.) chagasi compared with non-infected dogs (NID). The ID was grouped according to the clinical form and the parasite load. Symptomatic dogs (SD) presented higher parasite load in relation to oligosymptomatic (OD) and asymptomatic (AD). The qualitative expression of iNOS was observed only in ID. SD presented strong and prominent labeling of iNOS, followed by OD and AD. Quantitatively, the results showed that the median expression of iNOS was higher in SD and OD compared to NID. Also, dog spleens with high parasitism load showed marked iNOS expression. Taken together, the results suggest that the expression of iNOS in the spleen of infected dogs with CVL was associated with clinical worsening of the disease and with high parasitism.

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References

  • Alvar J, Canavate C, Molina R, Moreno J, Nieto J (2004) Canine leishmaniasis. Adv Parasitol 57:1–88

    Article  PubMed  Google Scholar 

  • Alves CF, Amorim IFG, Moura EP et al (2009) Expression of IFN-g, TNF-a, IL-10 and TGF-b in lymph nodes associates with parasite load and clinical form of disease in dogs naturally infected with Leishmania (Leishmania) chagasi. Vet Immunol Immunopathol 128:349–358

    Article  PubMed  CAS  Google Scholar 

  • Anderson CF, Mendez S, Sacks DL (2005) Nonhealing infection despite Th1 polarization produced by a strain of Leishmania major in C57BL/6 mice. J Immunol 174:2934–2941

    PubMed  CAS  Google Scholar 

  • Anstead GM, Chandrasekar B, Zhao W, Yang J, Perez LE, Melby PC (2001) Malnutrition alters the innate immune response and increases early visceralization following Leishmania donovani infection. Infect Immun 69:4709–4718

    Article  PubMed  CAS  Google Scholar 

  • Baptista-Fernandes T, Marques C, Roos Rodrigues O, Santos-Gomes GM (2007) Intra-specific variability of virulence in Leishmania infantum zymodeme MON-1 strains. Comp Immunol Microbiol Infect Dis 30:41–53

    Article  PubMed  Google Scholar 

  • Barbiéri CL (2006) Immunology of canine leishmaniasis. Parasite Immunol 28:329–337

    Article  PubMed  Google Scholar 

  • Barral-Netto M, Da Silva JS, Barral A, Reed S (1995) Up-regulation of T helper 2 and down-regulation of T helper 1 cytokines during murine retrovirus-induced immunodeficiency syndrome enhances susceptibility of a resistant mouse strain to Leishmania amazonensis. Am J Pathol 146:635–642

    PubMed  CAS  Google Scholar 

  • Belkaid Y, Hoffmann KF, Mendez S et al (2001) The role of interleukin (IL)-10 in the persistence of Leishmania major in the skin after healing and the therapeutic potential of anti-IL-10 receptor antibody for sterile cure. J Exp Med 194:1497–1506

    Article  PubMed  CAS  Google Scholar 

  • Bogdan C (2001) Nitric oxide and the immune response. Nat Immunol 2:907–916

    Article  PubMed  CAS  Google Scholar 

  • Brendolan A, Rosado MM, Carsetti R, Selleri L, Dear TN (2007) Development and function of the mammalian spleen. Bioessays 29:166–177

    Article  PubMed  CAS  Google Scholar 

  • Correa AP, Dossi AC, De Oliveira Vasconcelos R, Munari DP, De Lima VM (2007) Evaluation of transformation growth factor beta 1, interleukin-10, and interferon-gamma in male symptomatic and asymptomatic dogs naturally infected by Leishmania (Leishmania) chagasi. Vet Parasitol 143:267–274

    Article  PubMed  CAS  Google Scholar 

  • De Moura TR, Oliveira F, Novais FO et al (2007) Enhanced Leishmania braziliensis infection following pre-exposure to sandfly saliva. PLoS Negl Trop Dis 28:84

    Article  Google Scholar 

  • Fowell DJ, Locksley RM (1999) Leishmania major infection of inbred mice: unmasking genetic determinants of infectious diseases. Bioessays 21:510–518

    Article  PubMed  CAS  Google Scholar 

  • Gantt KR, Schultz-Cherry S, Rodriguez N et al (2003) Activation of TGF-beta by Leishmania chagasi: importance for parasite survival in macrophages. J Immunol 170:2613–2620

    PubMed  CAS  Google Scholar 

  • Gontijo CMF, Melo MN (2004) Leishmaniose visceral no Brasil: quadro atual, desafios e perspectivas. Rev Bras Epid 7:338–349

    Article  Google Scholar 

  • Hassan MF, Zhang Y, Engwerda CR, Kaye PM, Sharp H, Bickle QD (2006) The Schistosoma mansoni hepatic egg granuloma provides a favorable microenvironment for sustained growth of Leishmania donovani. Am J Pathol 169:943–953

    Article  PubMed  CAS  Google Scholar 

  • Lage RS, Oliveira GC, Busek SU et al (2007) Analysis of the cytokine profile in spleen cells from dogs naturally infected by Leishmania chagasi. Vet Immunol Immunopathol 115:135–145

    Article  PubMed  CAS  Google Scholar 

  • Li H, Poulos TL (2005) Structure–function studies on nitric oxide synthases. J Inorg Biochem 99:293–305

    Article  PubMed  CAS  Google Scholar 

  • Liew FY, Wei XQ, Proudfoot L (1997) Cytokines and nitric oxide as effector molecules against parasitic infections. Philos Trans R Soc Lond B Biol Sci 352:1311–1315

    Article  PubMed  CAS  Google Scholar 

  • Lima WG, Oliveira PS, Caliari MV et al (2007) Histopathological and immunohistochemical study of type 3 complement receptors (CD11b/CD18) in livers and spleens of asymptomatic and symptomatic dogs naturally infected with Leishmania (Leishmania) chagasi. Vet Immunol Immunopathol 117:129–136

    Article  PubMed  CAS  Google Scholar 

  • Lipoldová M, Svobodová M, Krulová M et al (2000) Susceptibility to Leishmania major infection in mice: multiple loci and heterogeneity of immunopathological phenotypes. Genes Immun 1:200–206

    Article  PubMed  Google Scholar 

  • Mancianti F, Gramiccia M, Gradoni L, Pieri S (1988) Studies on canine leishmaniasis control. 1. Evolution of infection of different clinical forms of canine leishmaniasis following antimonial treatment. Trans Roy Soc Trop Med Hyg 82:566–567

    Article  PubMed  CAS  Google Scholar 

  • Manna L, Reale S, Viola E, Vitale F, Manzillo VF, Michele PL, Caracappa S, Gravino AE (2006) Leishmania DNA load and cytokine expression levels in asymptomatic naturally infected dogs. Vet Parasitol 20:271–280

    Article  Google Scholar 

  • Marletta MA (1988) Mammalian synthesis of nitrite, nitrate, nitric oxide, and N-nitrosating agents. Chem Res Tox 1:249–257

    Article  CAS  Google Scholar 

  • Mbow ML, Bleyenberg JA, Hall LR, Titus RG (1998) Phlebotomus papatasi sand fly salivary gland lysate down-regulates a Th1, but up-regulates a Th2, response in mice infected with Leishmania major. J Immunol 161:5571–5577

    PubMed  CAS  Google Scholar 

  • Moreno J, Alvar J (2002) Canine leishmaniasis: epidemiological risk and the experimental model. Trends Parasitol 18:399–405

    Article  PubMed  Google Scholar 

  • Oliveira F, Lawyer PG, Kamhawi S, Valenzuela JG (2008) Immunity to distinct sand fly salivary proteins primes the anti-leishmania immune response towards protection or exacerbation of disease. PLoS Negl Trop Dis 16:e226

    Article  Google Scholar 

  • Olivier M, Gregory DJ, Forget G (2005) Subversion mechanisms by which Leishmania parasites can escape the host immune response: a signaling point of view. Clin Microbiol Rev 18:293–305

    Article  PubMed  CAS  Google Scholar 

  • Pacher P, Beckman JS, Liaudet L (2007) Nitric oxide and peroxynitrite in health and disease. Physiol Rev 87:315–424

    Article  PubMed  CAS  Google Scholar 

  • Palatnik-De-Souza CB, Santos WR, França-Silva JC, Costa RT, Reis AB, Palatnik M, Mayrink W, Genaro O (2001) Impact of canine control in the epidemiology of canine and human visceral leishmaniasis in Brazil. Am J Trop Med Hyg 65:510–517

    Google Scholar 

  • Pinelli E, Gebhard D, Mommaas AM et al (2000) Infection of a canine macrophage cell line with Leishmania infantum: determination of nitric oxide production and anti-leishmanial activity. Vet Parasitol 92:181–189

    Article  PubMed  CAS  Google Scholar 

  • Proudfoot L, Nikolaev AV, Feng GJ et al (1996) Regulation of the expression of nitric oxide synthase and leishmanicidal activity by glycoconjugates of Leishmania lipophosphoglycan in murine macrophages. Proc Natl Acad Sci USA 1:10984–10989

    Article  Google Scholar 

  • Reis AB, Martins-Filho OA, Teixeira-Carvalho A et al (2006) Parasite density and impaired biochemical/hematological status are associated with severe clinical aspects of canine visceral leishmaniasis. Res Vet Sci 81:68–75

    Article  PubMed  CAS  Google Scholar 

  • Rodrigues CA, Batista LF, Teixeira MC, Pereira AM, Santos PO, De Sa Oliveira GG, De Freitas LA, Veras PS (2007) Peripheral blood mononuclear cell supernatants from asymptomatic dogs immunized and experimentally challenged with Leishmania chagasi can stimulate canine macrophages to reduce infection in vitro. Vet Parasitol 143:197–205

    Article  PubMed  CAS  Google Scholar 

  • Rodríguez-Sosa M, Rivera-Montoya I, Espinoza A et al (2006) Acute cysticercosis favours rapid and more severe lesions caused by Leishmania major and Leishmania mexicana infection, a role for alternatively activated macrophages. Cell Immunol 242:61–71

    Article  PubMed  Google Scholar 

  • Sanchez MA, Diaz NL, Zerpa O, Negron E, Convit J, Tapia FJ (2004) Organ-specific immunity in canine visceral leishmaniasis: analysis of symptomatic and asymptomatic dogs naturally infected with Leishmania chagasi. Am J Trop Med Hyg 70:618–624

    PubMed  Google Scholar 

  • Santana CC, Vassallo J, De Freitas LA et al (2008) Inflammation and structural changes of splenic lymphoid tissue in visceral leishmaniasis: a study on naturally infected dogs. Parasite Immunol 30:515–24

    Article  PubMed  CAS  Google Scholar 

  • Solano-Gallego L, Koutinas A, Miró G et al (2009) Directions for the diagnosis, clinical staging, treatment and prevention of canine leishmaniosis. Vet Parasitol 165(1–2):1–18

    Article  PubMed  CAS  Google Scholar 

  • Stenger S, Thuring H, Rollinghoff M, Bogdan C (1994) Tissue expression of inducible nitric oxide synthase is closely associated with resistance to Leishmania major. J Exp Med 180:783–793

    Article  PubMed  CAS  Google Scholar 

  • Tasca KI, Buzetti WA, Tenorio MDAS et al (2009) Parasitological, immunohistochemical and histopathological study for Leishmania chagasi detection in splenic tissues of dogs with visceral leishmaniasis. Rev Bras Parasitol Vet 18:27–33

    PubMed  Google Scholar 

  • Wilson ME, Jeronimo SM, Pearson RD (2005) Immunopathogenesis of infection with the visceralizing Leishmania species. Microb Pathog 38:147–160

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank CAPES (Coordenação de Aperfeiçoamento de Pessoal de Nível Superior), NUPEB (Núcleo de Pesquisas em Ciências Biológicas) at Universidade Federal de Ouro Preto and Professor Vinicius Vieira Vignoli (UNIFENAS) for language review.

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Authors and Affiliations

  1. Universidade Presidente Antonio Carlos, Teófilo Otoni, MG, Brazil

    Fernando Rocha dos Santos

  2. Laboratório de Imunopatologia, NUPEB, Universidade Federal de Ouro Preto, Ouro Preto, MG, Brazil

    Paula Melo Abreu Vieira, Rodolfo Cordeiro Giunchetti, Claudia Martins Carneiro & Alexandre Barbosa Reis

  3. Laboratório de Imunologia Celular e Molecular, Instituto René Rachou—FIOCRUZ, Belo Horizonte, MG, Brazil

    Rodrigo Correa-Oliveira

  4. Laboratório de Microbiologia e Imunologia Básicas, Universidade Federal de Alfenas, Alfenas, MG, Brazil

    Luiz Cosme Cotta Malaquias

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  1. Fernando Rocha dos Santos
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  2. Paula Melo Abreu Vieira
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  3. Rodrigo Correa-Oliveira
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  7. Luiz Cosme Cotta Malaquias
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Correspondence to Luiz Cosme Cotta Malaquias.

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dos Santos, F.R., Vieira, P.M.A., Correa-Oliveira, R. et al. Qualitative and quantitative immunohistochemical evaluation of iNOS expression in the spleen of dogs naturally infected with Leishmania chagasi . Parasitol Res 108, 1397–1403 (2011). https://doi.org/10.1007/s00436-010-2183-5

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  • DOI: https://doi.org/10.1007/s00436-010-2183-5

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