Comparision of the Effects of Leishmania Soluble Antigen (LSA) and Lipopolysaccharide (LPS) on C57BL/6 Mice Macrophage Function

Document Type : Original Article

Authors

1 Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran.

2 Department of Immunology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.

Abstract

Background: Macrophages activation is the important anti-leishmania immune response. Different signals could affect macrophages development and functional activation. In the present study, we compared the effect of Leishmania Soluble Antigen (LSA) and Lipopolysaccharide (LPS) on peritoneal macrophage responses. Appropriate activation of macrophages depends on the signals they receive from pathogens and their different functional differentiation is crucial for anti-leishmania effects of macrophages.
Materials and Methods: In order to assay C57BL/6 mice macrophage function after LPS or LSA treatment, we measured phagocytic activity, cytokine pattern, and Nitric Oxide (NO) production by macrophages.
Results: Phase contrast microscopy showed that LPS-treated macrophages became more granular and spindle-shaped and similar to untreated macrophages, LSA-treated cells displayed round and spindle-shaped morphology. In addition, Nitric oxide assay and cytokine analysis showed that IL-6, IL-10, and TNF-α production was significantly reduced in LSA-treated macrophages in comparison with LPS-stimulated cells. It was also found that LSA-treated macrophages represented an anti-inflammatory phenotype compared with LPS-treated macrophages.
Conclusion: This anti-inflammatory phenotype was related to increase in IL-10/TNF- α production of LSA-treated macrophages and there was no difference in the amount of TGF-β between LSA- and LPS-treated groups.

Keywords


Yao C, Donelson JE, Wilson ME. The major surface protease (MSP or GP63) of Leishmania sp. Biosynthesis, regulation of expression, and function. Molecular and Biochemical Parasitology. 2003; 132(1):1–16. [DOI:10.1016/S0166-6851(03)00211-1]
Naderer T, McConville MJ. The Leishmania-macrophage interaction: A metabolic perspective. Cellular Microbiology. 2008; 10(2):301–8. [DOI:10.1111/j.1462-5822.2007.01096.x]
Iniesta V, Gómez Nieto LC, Molano I, Mohedano A, Carcelén J, Mirón C, et al. Arginase I induction in macrophages, triggered by Th2-type cytokines, supports the growth of intracellular Leishmania parasites. Parasite Immunol. 2002; 24(3):113–8. [DOI:10.1046/j.1365-3024.2002.00444.x] [PMID]
Iniesta V, Carcelén J, Molano I, Peixoto PM V, Redondo E, Parra P, et al. Arginase I induction during Leishmania major infection mediates the development of disease. Infect Immun. 2005; 73(9):6085–90. [DOI:10.1128/IAI.73.9.6085-6090.2005] [PMID] [PMCID]
Kropf P, Fuentes JM, Fähnrich E, Arpa L, Herath S, Weber V, et al. Arginase and polyamine synthesis are key factors in the regulation of experimental leishmaniasis in vivo. FASEB J. 2005; 25(2):1000–2. [DOI:10.1096/fj.04-3416fje] [PMID]
Ferraz Coelho EA, Pereira Tavares CA, Amorim Carvalho FA, Chaves KF, Teixeira KN, Rodrigues RC, et al. Immune responses induced by the Leishmania (Leishmania) donovani A2 antigen, but not by the LACK antigen, are protective against experimental Leishmania (Leishmania) amazonensis infection. Infect Immun. 2003; 71(7):3988–94. [DOI:10.1128/IAI.71.7.3988-3994.2003] [PMCID]
Williams LM, Ridley AJ. Lipopolysaccharide Induces Actin Reorganization and Tyrosine Phosphorylation of Pyk2 and Paxillin in Monocytes and Macrophages. 2000; 164(4):2028-36.
Kleveta G, Borzȩcka K, Zdioruk M, Czerkies M, Kuberczyk H, Sybirna N, et al. LPS induces phosphorylation of actin-regulatory proteins leading to actin reassembly and macrophage motility. J Cell Biochem. 2012; 113(1):80–92. [DOI:10.1002/jcb.23330] [PMID]
Soudi S, Zavaran-Hosseini A, Muhammad Hassan Z, Soleimani M, Jamshidi Adegani F, Hashemi SM. Comparative study of the effect of LPS on the function of BALB/c and C57BL/6 peritoneal macrophages. Cell Journal. 2013; 15(1):45–54. [PMID] [PMCID]
Rossol M, Heine H, Meusch U, Quandt D, Klein C, Sweet MJ, et al. LPS-induced Cytokine Production in Human Monocytes and Macrophages. Crit Rev Immunol. 2011; 31(5):379–446. [DOI:10.1615/CritRevImmunol.v31.i5.20] [PMID]
Dobrovolskaia MA, Vogel SN. Toll receptors, CD14, and macrophage activation and deactivation by LPS. Microbes and Infection. 2002; 4(9):903-14. [DOI:10.1016/S1286-4579(02)01613-1]
Bode JG, Ehlting C, Häussinger D. The macrophage response towards LPS and its control through the p38MAPK–STAT3 axis. Cellular signalling. 2012; 24(6):1185-94. [DOI:10.1016/j.cellsig.2012.01.018] [PMID]
Balzola F, Bernstein C, Ho GT, Russell RK. TLR signalling augments macrophage bactericidal activity through mitochondrial ROS: Commentary. Inflammatory Bowel Disease Monitor. 2011; 472(7344):476-80.
Martinez FO, Gordon S. The M1 and M2 paradigm of macrophage activation: time for reassessment. F1000Prime Rep. 2014; 6(13):13. [DOI:10.12703/P6-13]
Murray PJ, Wynn T a. Protective and pathogenic functions of macrophage subsets. Nat Rev Immunol. Nature Publishing Group. 2011; 11(11):723–37.
PromoCell GMBH, Note A. Differentiation of M1- or M2-Macrophages from PBMC / Monocytes. 2013; 1-9.
Vogel DYS, Glim JE, Stavenuiter AWD, Breur M, Heijnen P, Amor S, et al. Human macrophage polarization in vitro: Maturation and activation methods compared. Immunobiology. 2014;219(9):695–703. [DOI:10.1016/j.imbio.2014.05.002] [PMID]
Sica A, Mantovani A. Macrophage plasticity and polarization: in vivo veritas. The Journal of clinical investigation. 2012; 122(3):787-95. [DOI:10.1172/JCI59643] [PMID] [PMCID]
MacMicking J, Xie QW, Nathan C. Nitric oxide and macrophage function. Annu Rev Immunol. 1997; 15(1):323–50. [DOI:10.1146/annurev.immunol.15.1.323] [PMID]
MacMicking J, Xie Q, Nathan C. NITRIC OXIDE AND MACROPHAGE FUNCTION. Annu Rev Immunol. 1997; 15(1):323–50. [DOI:10.1146/annurev.immunol.15.1.323] [PMID]
Chanteux H, Guisset AC, Pilette C, Sibille Y. LPS induces IL-10 production by human alveolar macrophages via MAPKinases- and Sp1-dependent mechanisms. Respir Res. 2007; 8(1):71. [DOI:10.1186/1465-9921-8-71] [PMID] [PMCID]
Couper K, Blount D, Riley E. IL-10: the master regulator of immunity to infection. J Immunol. 2008; 180(9):5771–7. [DOI:10.4049/jimmunol.180.9.5771] [PMID]
Alderton WK, Cooper CE, Knowles RG. Nitric oxide synthases: structure, function and inhibition. Biochem J. 2001; 357(Pt3):593–615. [DOI:10.1042/0264-6021:3570593] [PMID] [PMCID]