Parasites, Hosts and Diseases

The Korea Society for Parasitology and Tropical Medicine (대한기생충학·열대의학회)
권호 표지

Production of nitric oxide by murine macrophages induced by lipophosphoglycan of Leishmania major

  • KAVOOSI Gholamreza (Institute of Biochemistry and Biophysics, University of Tehran) ;
  • ARDESTANI Sussan K. (Institute of Biochemistry and Biophysics, University of Tehran) ;
  • KARIMINIA Amina (Department of Immunology, Pasteur Institute of Iran) ;
  • TAVAKOLI Zahra (Institute of Biochemistry and Biophysics, University of Tehran)
  • Published : 2006.03.01
Download PDF
⟨ Previous Next ⟩

Abstract

Protozoan parasites of the genus Leishmania cause a number of important human diseases. One of the key determinants of parasite infectivity and survival is the surface glycoconjugate lipophosphoglycan (LPG). In addition, LPG is shown to be useful as a transmission blocking vaccine. Since culture supernatant of parasite promastigotes is a good source of LPG, we made attempts to characterize functions of the culture supernatant, and membrane LPG isolated from metacyclic promastigotes of Leishmania major. The purification scheme included anion-exchange chromatography, hydrophobic interaction chromatography and cold methanol precipitation. The purity of supernatant LPG (sLPG) and membrane LPG (mLPG) was determined by SOS-PAGE and thin layer chromatography. The effect of mLPG and sLPG on nitric oxide (NO) production by murine macrophages cell line (J77 4.1 A) was studied. Both sLPG and mLPG induced NO production in a dose dependent manner but sLPG induced significantly higher amount of NO than mLPG. Our results show that sLPG is able to promote NO production by murine macrophages.

Keywords

References

  1. Ames BN (1966) Assay of inorganic phosphate, total phosphate and phosphatases. Methods in Enzymol 8: 115-118 https://doi.org/10.1016/0076-6879(66)08014-5
  2. Balararnan S, Tewary P, Singh VK, Madhubala R (2004) Leishmania donovani induces interferon regulatory factor in murine macrophages: a host defense response. Biochem Biophys Res Commun 317: 639-647 https://doi.org/10.1016/j.bbrc.2004.03.097
  3. Becker I, Salaiza N, Aguirre M, Delagado J, CarrilloCarrasco N, Kobeh LG, Ruiz A, Cervantes R, Torres AP, Cabrera N, Gonzalez A, Maldonado C, Isibasi A (2003) Leishmania lipophosphoglycan activates NK cells through toll-like receptor-2. Mol Biochem Parasitol 130: 65-74 https://doi.org/10.1016/S0166-6851(03)00160-9
  4. Cunningham AC (2002) Parasitic adaptive mechanisms in infection by Leishmania. Exp Mol Pathol 72: 132-41 https://doi.org/10.1006/exmp.2002.2418
  5. Davies CR, Cooper AM, Peacock C, Lane RP, Blackwell JM (1990) Expresion of LPG and GP63 by different developmental stages of Leishmania major in the sand fly Phlebotomus papatasi. Parasitology 101: 337-343 https://doi.org/10.1017/S0031182000060522
  6. de Veer MJ, Curtis JM, Baldwin TM, DiDonato JA, Sexton A, McConville MJ, Handman E, Schofield L (2003) MyD88 is essential for clearance of Leishmania major: possible role for lipophosphoglycan and toll-like receptore 2 signalling. Eur J Immunol 33: 2822-2831 https://doi.org/10.1002/eji.200324128
  7. Descoteaux A, Turco SJ (2002) Functional aspects of the Leishmania donovani lipophosphoglycan during macrophage function. Microbes Infect 4: 975-981 https://doi.org/10.1016/S1286-4579(02)01624-6
  8. Handman E, Bullen DV (2002) Interaction of Leishmania with host macrophage. Trends Parasitol 18: 332-334 https://doi.org/10.1016/S1471-4922(02)02352-8
  9. Ilg T, Stierhof YD, Craik D, Simpson R, Handman E, Bacic A (1996) Purification and structural characterization of a filamentous, mucinlike proteophosphoglycan secreted by Leishmania parasites. J Biol Chem 271: 21583-21596 https://doi.org/10.1074/jbc.271.35.21583
  10. Ilg T, Stierhof YD, Wiese M, McConville MJ, Overath P (1994) Characterization of phosphoglycan-containing secretory products of Leishmania. Parasitology 108: S63-71 https://doi.org/10.1017/S0031182000075739
  11. 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 https://doi.org/10.1098/rstb.1997.0115
  12. McConville MJ, Bacic A (1989) A family of glycoinositol phospholipids from Leishmania major. Isolation, characterization, and antigenicity. J Biol Chem 264: 757-766
  13. McConville MJ, Turco SJ, Ferguson MA, Sacks DL (1992) Developmental modification of lipophosphoglycan during the differentiation of Leishmania major promastigotes to an infectious stage. EMBO J 11: 3593-3600
  14. Miranda MK, Espey MG, Wink DA (2001) A rapid, simple spectrophotometric method for simultaneous detection of nitrate and nitrite. Nitric Oxide 5: 62-71 https://doi.org/10.1006/niox.2000.0319
  15. 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 https://doi.org/10.1128/CMR.18.2.293-305.2005
  16. Proudfoot L, Nikoleav AV, Feng GJ, Wei WQ, Ferguson MA, Brimocombe JS, Liew FY (1996) Regulation of the expression of nitric oxide synthase and leishmaniacidal activity by glycoconjugates of Leishmaina lipophosphoglycan in murine macrophages. Proc Natl Acad Sci USA 93: 10984-10989
  17. Proudfoot L, O'Donnell CA, Liew FY (1995) Glycoinositolphospholipids of Leishmania major inhibit nitric oxide synthesis and reduce leishmaniacidal activity in murine macrophages. Eur J Immunol 25: 745-750 https://doi.org/10.1002/eji.1830250318
  18. Rao P, Pattabiraman TN (1989) Reevaluation of the phenolsulfuric acid reaction for the estimation of hexoses and pentoses. Anal Biochem 181: 18-22 https://doi.org/10.1016/0003-2697(89)90387-4
  19. Ropert C, Gazzinelli RT (2000) Signaling of immune system cells by glycosylphosphatidylinositol (GPI) anchor and related structures derived from parasitic protozoa. Curr Opin Microbial 3: 395-403 https://doi.org/10.1016/S1369-5274(00)00111-9
  20. Sacks DL (1989) Metacyclogenesis in Leishmania amastigotes. Exp Parasitol 69: 100-103 https://doi.org/10.1016/0014-4894(89)90176-8
  21. Tonui WK, Mbati P A, Anjili CO, Orago AS, Turco SJ, Githure JI, Koech DK (2001) Transmission blocking vaccine studies in leishmaniasis: I. Lipophosphoglycan is a promising transmission blocking vaccine molecule against cutaneous leishmaniasis. East Afr Med J 78: 84-89
  22. Tsai CM, Frasch CE (1982) A sensitive silver stain for detecting lipopolysaccharide in polyacrylamide gels. Anal Biachem 119: 115-119 https://doi.org/10.1016/0003-2697(82)90673-X
  23. Turco SJ, Descoteaux A (1992) The lipophosphoglycan of Leishmania parasites. Annu Rev Microbiol 46: 65-94 https://doi.org/10.1146/annurev.mi.46.100192.000433