Leishmania Donovani Cell Surface Sialoglycans Regulate Susceptibility for Siglec Mediated Macrophage Invasion and Parasite Survival

Subir Karmakar, Siddhartha K Bhaumik, Joydeep Paul, Tripti De


Glycoconjugates play a pivotal role in the survival of Leishmania parasites in destructive surroundings. An important constituent present on many glycoconjugates is sialic acid. By virtue of their peripheral position on oligosaccharide chains of glycoconjugates, sialic acids are well suited as molecular determinants of specific biological processes, including the interaction of pathogenic microorganisms with sialylated cellular receptors. Differences in a2,3- and a2,6-sialoglycan patterns detected in clonal virulent Leishmania donovani promastigotes, correlated with the level of a2,3- and a2,6-sialyltransferase activity present in these parasites. The role of macrophage sialic acid-receptors in uptake and survival of L.donovani was studied in the murine macrophage cell line raw 264.7. Macrophage invasion was dependent on the binding to Siglec-1, while suppression of MAPK signaling was mediated through Siglec-5. Sialic acid removal by neuraminidase treatment reduced parasite infectivity. The presence of trypsin resistant sialic acid residues in the neuraminidase treated parasites grown in a serum free medium in presence of sialoglycoconjugates indicated that the parasites could salvage sialic acid from exogenous sialoglycans and reutilize it for de novo glycoprotein sialylation in L.donovani parasites. Thus, our results demonstrate the involvement of sialoglycans in the invasion as well as the survival process of L.donovani parasites.


Leishmania donovani; sialoglycans; macrophage; sialoadhesine; p38 MAPK; ERK; JNK


Abu-Dayyeh I, Shio MT, Sato S, Akira S, Cousineau B & Oliver M 2008 Leishmania-induced IRAK-1 inactivation is mediated by SHP-1 interacting with an evolutionary con- served KTIM motif. Plos Negl Trop Dis 2 e305.

Angata T & Varki A 2000 Siglec-7: a sialic acid-binding lectin of the immunoglobulin superfamily. Glycobiology 10 431-438.

Angata T 2006 Molecular diversity and evolution of the siglec family of cell surface lectins. Mol Divers 10 555-566.

Appelmelk BJ, van Die I, van Vliet SJ, Vandenbroucke- Grauls CM, Geitenbeek TB & van Kooyk Y 2003 Carbohydrate profiling identifies new pathogens that interact with dendritic cell-specific ICAM-3-grabbing nonintegrin on dendritic cells. J Immunol 170 1635-1639.

Aumiller JJ, Hollister JR & Jarvis DL 2003 A transgenic insect cell line engineered to produce CMP–sialic acid and sialylated glycoproteins. Glycobiology 13 497-507.

d’Avila-Levy CM, Araújo FM, Vermelho AB, Branquinha MH, Alviano CS, Soares RM & dos Santos AL 2004 Differential lectin recognition of glycoproteins in choanomastigote-shaped trypanosomatids: taxonomic implications. FEMS Microbiol Lett 231 171-176.

Azuma Y, Sakanashi M & Matsumoto K 2001 The effect of alpha2,6-linked sialic acid on anti-IgM antibody-induced apoptosis in Ramos cells. Glycoconj J 18 419-424.

Basu M, Basu S, Stoffyn A & Stoffyn P 1982 Biosynthesis in vitro of sialyl(a2-3)neolactotetraosylceramide by a sialyl- transferase from embryonic chicken brains. JBiolChem 257 12765-12769.

Ben-Othman R, Guizani-Tabbane L & Dellagi K 2008 Leishmania initially activates but subsequently down regulates intracellular mitogen-activated protein kinases and nuclear factor kappaB signaling in macrophages. Mol Immunol 45 3222-3229.

Berninsone P, Eckhardt M, Gerardy-Schahn R & Hirschberg CB 1997 Functional expression of the murine Golgi CMP-sialic acid transporter in Saccharomyces cerevisiae. J Biol Chem 272 12616-12619.

Bhaumik SK, Singh M, Basu R, Bhaumik S, Roychoudhury K, Naskar K, Roy S & De T 2008 Virulence attenuation of a UDP-galactose/N-acetylglucosamine beta1,4- galactosyltransferase expressing Leishmania donovani promastigote. Glycoconj J 25 459-472.

Blanchette J, Racette N, Faure R, Siminovitch KA & Olivier M 1999 Leishmania-induced increase activation of macrophage SHP-1 tyrosine phosphatase are associated with impaired IFN-gamma triggered JAK2 activation. Eur J Immunol 29 3737-3744.

Brinkman-Van der Linden EC, Angata T, Reynolds SA, Powell LD, Hendrick SM & Varki A 2003 CD33/Siglec-3 binding specificity, expression pattern, and consequences of gene deletion in mice. Mol Cell Biol 23 4199-4206.

Buschiazzo A, Tavares GA, Campetella O, Spinelli S, Cremona ML, París G, Amaya MF, Frasch AC & Alzari PM 2000 Structural basis of sialyltransferase activity in trypanosomal sialidases. EMBO J 19 16-24.

Butcher BA, Sklar LA, Seamer LC & Glew RH 1992 Heparin enhances the interaction of infective Leishmania donovani promastigotes with mouse peritoneal macrophages A fluorescence flow cytometric analysis. J Immunol 148 2879- 2886.

Chang KP 1981 Leishmania donovani-macrophage binding mediated by surface glycoproteins/antigens:characterization in vitro by a radioisotopic assay. Mol Biochem Parasitol 4 67-76.

Chatterjee M, Chava AK, Kohla G, Pal S, Merling A, Hin- derlich S, Unger U, Strasser P, Gerwig GJ, Kamerling JP, Vlasak R, Crocker PR, Schauer R, Schwartz-Albiez R & Mandal C 2003 Identification and characterization of adsorbed serum sialoglycans on Leishmania donovani promastigotes. Glycobiology 13 351-361.

Cheng J, Yu H, Lau K, Huang S, Chokhawala HA, Li Y, Tiwari VK & Chen X 2008 Multifunctionality of Campylobacter jejuni sialyltransferase CstII: Characterization of GD3/GT3 oligosaccharide synthase, GD3 oligosaccharide sialidase, and trans-sialidase activities. Glycobiology 18 686 -697.

Cheng J, Huang S, Yu H, Li Y, Lau K & Chen X 2010 Transsialidase activity of Photobacterium damsela alpha2,6 -sialyltransferase and its application in the synthesis of sialosides. Glycobiology 20 260-268.

Chitnis CE 2001 Molecular insights into receptors used by malaria parasites for erythrocyte invasion. Curr Opin Hematol 8 85-91.

Colli W 1993 Trans sialidase: a unique enzyme activity discovered in the protozoan Trypanosoma cruzi. FASEB J 7 1257-1264.

Crocker PR & Varki A 2001 Siglecs in the immune system. Immunology 103 137-145.

Crocker R, Paulson JC & Varki A 2007 Siglecs and their role in the immune system. Nat Rev Immunol 7 255-266.

De T & Roy S 1999 Infectivity and attenuation of Leishmania donovani promastigotes: association of galactosyl transferase with loss of parasite virulence. J Parasitol 85 54-59.

De Souza W 1995 Structural organization of the surface of pathogenic protozoa. Micron 26 405-430.

Durasingh MT, Maier AG, Triqlia T & Cowman AF 2003 Erythrocyte-binding antigen 175 mediates invasion in Plasmodium falciparum utilizing sialic acid-dependent and -independent pathways. Proc Natl Acad Sci USA 100 4796- 4801.

Dwyer DM 1977 Leishmania donovani: surface membrane carbohydrates of promastigotes. Exp Parasitol 41 341-358.

Engstler M, Schauer R & Brun R 1995 Distribution of developmentally regulated trans-sialidase in the Kinetoplastida and characterization of a shed trans-sialidase from procyclic Trypanosoma congolense. Acta Trop 59 117-129.

Ferguson MA 1997 The surface glycoconjugates of trypanosomatid parasites. Philos Trans R Soc Lond B Biol Sci 352 1295-1302.

Garin YJ, Sulahian A, Pratlong F, Meneceur P, Gangneux JP, Prina E, Dedet JP & Derouin F 2001 Virulence of Leishmania infantum is expressed as a clonal and dominant phenotype in experimental infections. Infect Immun 69 7365 -7373.

Ghoshal A, Gerwig GJ, Kamerling JP & Mandal C 2010 Sialic acid in different Leishmania sp, its correlation with nitric oxide resistance and host responses. Glycobiology 20 553-566.

Guha-Niyogi A, Sullivan DR & Turco SJ 2001 Glycoconjugate structures of parasitic protozoa. Glycobiology 11 45R- 59R.

Harduin-Lepers A, Mollicone R, Delannoy P & Oril R 2005 The animal sialyltransferases and sialyltransferase-related genes: a phylogenetic approach. Glycobiology 15 805-817. Hager KM & Carruthers VB 2008 Marveling at parasite invasion. Trends Parasitol 24 51-54.

Harvey HA, Porat N, Campbell CA, Jennings M, Gibson BW, Phillips NJ, Apicella MA & Blake MS 2000 Gonococcal lipooligosaccharide is a ligand for the asialoglycoprotein receptor on human sperm. Mol Microbiol 36 1059-1070.

Hollister JR & Jarvis DL 2001 Engineering lepidopteran insect cells for sialoglycoprotein production by genetic transformation with mammalian beta 1,4- galactosyltransferase and alpha 2,6-sialyltransferase genes. Glycobiology 11 1-9.

Hollister JR, Grabenhorst E, Nimtz M, Condrat H & Jarvis DL 2002 Engineering the protein N-glycosylation pathway in insect cells for production of biantennary, complex N-glycans. Biochemistry 41 15093-15104.

Iovannisci DM, Plested CP & Moe GR 2010 Evidence for rosettes as an unrecognized stage in the life cycle of Leishmania parasites. J Eukaryot Microbiol 57 405-414.

Jacobs CL, Goon S, Yarema KJ, Hinderlich S, Hang HC, Chai DH & Bertozzi CR 2001 Substrate specificity of the sialic acid biosynthetic pathway. Biochemistry 40 12864- 12874.

Jeanneau C, Chazalet V, Auge C, Soumpasis DM, Harduin- Lepers A, Delannoy P, Imberty A & Brenton C 2004 Structure-function analysis of the human sialyltransferase ST3Gal I: role of n-glycosylation and a novel conserved sialylmotif. J Biol Chem 279 13461-13468.

Kamerling JP & Gerwig GJ 2006 Structural analysis of Naturally occuring sialic acids. Methods Mol Biol 347 69- 91.

Kamhawi S 2006 Phlebotomine sand flies and Leishmania parasites: friends or foes? Trends Parasitol 22 439-445.

Karmakar S, Paul J & De T 2011 Leishmania donovani glycosphingolipid facilitates antigen presentation by inducing relocation of CD1d into lipid rafts in infected macrophages. Eur J Immunol 41 1376-1387.

Lauc G & Heffer-Lauc M 2006 Shedding and uptake of gangliosides and glycophosphatidylinositol-anchored proteins. Biochim Biophys Acta 1760 584-602.

da Silva LM, Owens KL, Murta SM & Beverly SM 2009 Regulated expression of the Leishmania major surface virulence factor lipophosphoglycan using conditionally destabilized fusion proteins. Proc Natl Acad Sci USA 106 7583- 7588.

Manzi AE, Diaz S & Varki A 1990 High-pressure liquid chromatography of sialic acids on a pellicular resin anion- exchange column with pulsed amperometric detection: a comparison with six other systems. Anal Biochem 188 20- 32.

McConville MJ, Thomas-Oates JE, Ferguson MA & Homans SW 1990 Structure of the lipophosphoglycan from Leishmania major. J Biol Chem 265 19611-19623.

Monteiro VG, Soares CP, de Souza W 1998 Host cell surface sialic acid residues are involved on the process of penetration of Toxoplasma gondii into mammalian cells. FEMS Microbiol Lett 164 323-327.

Munday J, Floyd H & Crocker PR 1999 Sialic acid binding receptors (siglecs) expressed by macrophages. J Leukoc Biol 66 705-711.

Previato JO, Andrade AF, Pessolani MC & Mendonca-Previato L 1985 Incorporation of sialic acid into Trypanosoma cruzi macromolecules A proposal for a new metabolic route. Mol Biochem Parasitol 16 85-96.

Sacks DL, Hieny S & Sher A 1985 Identification of cell surface carbohydrate and antigenic changes between noninfective and infective developmental stages of Leishmania major promastigotes. J Immunol 135 564-569.

Schauer R, Reuter G, Mühlpfordt H, Andrade AF & Pereira ME 1983 The occurrence of N-acetyl- and N- glycolylneuraminic acid in Trypanosoma cruzi. Hoppe Seylers Z Physiol Chem 364 1053-1057.

Schauer R & Kamerling JP 1997 Chemistry, biochemistry and biology of sialic acids. In Glycoproteins II, pp 243-402. Eds J Montreuil, FG Vliegenthart & H Schachter. Amsterdam: Elsevier.

Schenkman RP, Vandekerckhoye F & Schenkman S 1993 Mammalian cell sialic acid enhances invasion by Trypanosoma cruzi. Infect Immun 61 898-902.

Schwarzmann G, Hoffman-Blehauer P, Schubert J, Sandhoff K & Marsh D 1983 Incorporation of ganglioside analogues into fibroblast cell membranes A spin-label study. Biochemistry 22 5041-5048.

Varki A & Diaz S 1984 The release and purification of sialic acids from glycoconjugates:methods to minimize the loss and migration of O-acetyl groups. Anal Biochem 137 236- 247.

Vannier-Santos MA, Saravia EM & de Souza W 1991 Nuclear and cytoplasmic lectin binding sites in promastigotes of Leishmania. J Histochem Cytochem 39 793-800.

Weinstein J, de Souza-e-Silva U & Paulson JC 1982 Purification of a Gal beta 1 to 4GlcNAc alpha 2 to 6 sialyltransferase and a Gal beta 1 to 3(4) GlcNAc alpha 2 to 3 sialyltransferase to homogeneity from rat liver. J Biol Chem 257 13835-13844.

Winter C, Schwegmann-Wessels C, Cavanagh D, Neumann U & Herrler G 2006 Sialic acid is a receptor determinant for infection of cells by avian infectious bronchitis virus. J Gen Virol 87 1209-1216.

Wlasichuk KB, Kashem MA, Nikard PV, Bird P, Jiang C & Venot AP 1993 Determination of the specificities of rat liver Gal(beta 1-4)GlcNAc alpha 2,6-sialyltransferase and Gal (beta 1-3/4)GlcNAc alpha 2,3-sialyltransferase using synthetic modified acceptors. J Biol Chem 268 13971-13977. Yu RK & Ledden RW 1972 Gangliosides of human, bovine, and rabbit plasma. J Lipid Res 13 680-686.

Yu H, Chokhawala H, Karpel R, Yu H, Wu B, Zhang J, Zhang Y, Jia Q & Chen X 2005 A multifunctional Pasteurella multocida sialyltransferase: A powerful tool for the synthesis of sialoside libraries. J Am Chem Soc 127 17618- 17619.

Zhao W, Chen TL, Vertel BM & Colley KJ 2006 The CMP- sialic acid transporter is localized in the medial-trans Golgi and possesses two specific endoplasmic reticulum export motifs in its carboxy-terminal cytoplasmic tail. J Biol Chem 281 31106-31118.

Full Text: PDF


  • There are currently no refbacks.

Copyright © 2018 Lorem Ipsum Press