Iranian Journal of Parasitology 2016. 11(3):325-331.

Optimization of the Timing of Induction for the Assessment of Nitric Oxide Production in Leishmania major Infected Macro-phage Cells
Somaye SADEGHI, Negar SEYED, Sima RAFATI, Tahereh TAHERI

Abstract


Background: The present study was conducted to investigate the optimized timing for macrophages induction and nitric oxide (NO) production against invading Leishmania parasite.

Methods: The present study examined the murine macrophage cell line, B10R, in three different states. In the first state, the cells were first infected with L. major and then treated with IFN-g and LPS as stimulants. In the second state, the cells were infected after stimulation with IFN-g and LPS. In the third state, the cells were only exposed to stimulants as controls. In all the three states, cell culture supernatants were collected at three points in time (6, 24 and 48 h) and the amount of NO production was measured using Griess assay.

Results: The treatment of macrophages with inducers prior to infection with stationary phase parasite led to the secretion of significant amounts of NO, particularly at early time points quit contrary to the cells infected with parasites prior to induction. The amount of NO produced by cells induced after infection was detected significantly lower.

Conclusion: The induction of macrophages prior to infection with parasites leads to the production and secretion of greater amounts of NO, resulting in an increased ability to suppress and inhibit parasite proliferation even in the early stages of infection.

Keywords


Leishmania major; Nitric oxide; Interferongamma; Lipopolysaccharide

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References


Gupta S. Visceral leishmaniasis: Experimental models for drug discovery. Indian J Med Res. 2011;133:27-39.

Colasanti M, Gradoni L, Mattu M, Persichini T, Salvati L, Venturini G, Ascenzi P. Molecular bases for the anti-parasitic effect of no (review). Int J Mol Med. 2002;9(2):131-4.

Croft SL, Seifert K, Yardley V. Current scenario of drug development for leishmaniasis. Indian J Med Res. 2006;123(3):399-410.

Shio MT, Olivier M. Editorial: Leishmania survival mechanisms: The role of host phosphatases. J Leukoc Biol. 2010;88:1-3.

Wei XQ, Charles IG, Smith A, Ure J, Feng GJ, Huang FP, Xu D, Muller W. Altered immune responses in mice lacking inducible nitric oxide synthase. Nature. 1995;375:408-11.

Dhingra N, Satapathy S. Altercating leishmaniasis: Cultivating improved vaccines. Immunol infect Dis. 2014;2:4-10.

Shio MT, Hassani K, Isnard A, Ralph B, Contreras I, Gomez MA, Abu-Dayyeh I, Olivier M. Host cell signalling and Leishmania mechanisms of evasion. J Trop Med. 2012;2012:819512. doi: 10.1155/2012/819512. Epub 2011 Nov 3.

Yang CS, Yuk JM, Jo EK. The role of nitric oxide in mycobacterial infections. Immune Netw. 2009;9(2):46-52.

Barak Y, Schreiber F, Thorne SH, Contag CH, Matin A. Role of nitric oxide in Salmonella typhimurium-mediated cancer cell killing. BMC Cancer. 2010;10:146.

Ascenzi P, Bocedi A, Gradoni L. The anti‐parasitic effects of nitric oxide. IUBMB Life. 2003;55(10-11):573-8.

Gutierrez FR, Mineo TW, Pavanelli WR, Guedes PM, Silva JS. The effects of nitric oxide on the immune system during Trypanosoma cruzi infection. Mem Inst Oswaldo Cruz. 2009;104(1):236-245.

Hämäläinen M. Inducible nitric oxide synthase as a target of anti-inflammatory treatment modalities [Dissertation]. University of Tampere, 2008.

James SL. Role of nitric oxide in parasitic infections. Microbiol Rev. 1995;59:533-547.

Li Y-H, Yan Z-Q, Brauner A, Tullus K. Activation of macrophage nuclear factor-kappa B and induction of inducible nitric oxide synthase by lps. Respir Res. 2002;3:23.

Doherty TM. T-cell regulation of macrophage function. Curr Opin Immunol. 1995;7(3):400-404.

Assreuy J, Cunha FQ, Epperlein M, Noronha‐Dutra A, O'Donnell CA, Liew FY, Moncada S. Production of nitric oxide and superoxide by activated macrophages and killing of Leishmania major. Eur J Immunol. 1994;24:672-676.

Balestieri FMP, Queiroz ARP, Scavone C, Costa VMA, Barral-Netto M, de Almeida Abrahamsohn I. Leishmania (l.) amazonensis-induced inhibition of nitric oxide synthesis in host macrophages. Microbes Infect. 2002;4(1):23-29.

Ota H, Takashima Y, Matsumoto Y, Hayashi Y, Matsumoto Y. Pretreatment of macrophages with the combination of IFN-gamma. And IL-12 induces resistance to Leishmania major at the early phase of infection. J Vet Med Sci. 2008;70(6):589-593.

Akilov OE, Kasuboski RE, Carter CR, McDowell MA. The role of mannose receptor during experimental leishmaniasis. J Leukoc Biol. 2007;81(5):1188-96.

Schlesinger L, Horwitz M. Phagocytosis of Mycobacterium leprae by human monocyte-derived macrophages is mediated by complement receptors cr1 (cd35), cr3 (cd11b/cd18), and cr4 (cd11c/cd18) and IFN-gamma activation inhibits complement receptor function and phagocytosis of this bacterium. J Immunol. 1991;147(6):1983-94.

Badaró R, Johnson WD. The role of interferon-γ in the treatment of visceral and diffuse cutaneous leishmaniasis. J Infect Dis. 1993;167:S13-S17.

Kiderlen AF, Lohmann-Matthes M-L. Treatment of experimental cutaneous and visceral murine leishmaniasis with recombinant gamma-interferon. Leishmaniasis. The Current Status and New Strategies for Control, Hart DT (ed.), Plenum Press, New York, NATO-ASI Series A163:457-466.

Zhong GM, De La Maza L. Activation of mouse peritoneal macrophages in vitro or in vivo by recombinant murine gamma interferon inhibits the growth of Chlamydia trachomatis serovar L1. Infect Immun. 1988;56(12):3322-25.

Noda T, Amano F. Differences in nitric oxide synthase activity in a macrophage-like cell line, Raw264.7 cells, treated with lipopolysaccharide (LPS) in the presence or absence of interferon-γ (IFN-γ): Possible heterogeneity of iNOS activity. J Biochem. 1997;121(1):38-46.

Heinzel FP, Sadick MD, Holaday BJ, Coffman R, Locksley RM. Reciprocal expression of interferon gamma or interleukin 4 during the resolution or progression of murine leishmaniasis. Evidence for expansion of distinct helper t cell subsets. J Exp Med. 1989;169(1):59-72.

Locksley RM, Heinzel FP, Sadick MD, Holaday BJ, Gardner KD. Murine cutaneous leishmaniasis: Susceptibility correlates with differential expansion of helper t-cell subsets. Ann Inst Pasteur Immunol. 1987;138(5):744-9.

Panosian CB, Wyler DJ. Acquired macrophage resistance to in vitro infection with Leishmania. J Infect Dis. 1983;148(6):1049-1054.

Hoff R. Killing in vitro of Trypanosoma cruzi by macrophages from mice immunized with T. cruzi or BCG, and absence of cross-immunity on challege in vivo. J Exp Med. 1975;142:299-311.

Nacy CA, Meltzer MS. Macrophages in resistance to rickettsial infections: Protection against lethal Rickettsia tsutsugamushi infections by treatment of mice with macrophage-activating agents. J Leukoc Biol. 1984;35:385-396.

Horwitz MA, Silverstein SC. Activated human monocytes inhibit the intracellular multiplication of Legionnaires' disease bacteria. J Exp Med. 1981;154(5):1618-1635.


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