Targeted Gene Deletion of Leishmania major UDP-galactopyranose Mutase Leads to Attenuated Virulence

Considering the high incidence of galactofuranose (Galf) in pathogens and its absence from higher eukaryotes, the enzymes involved in the biosynthesis of this unusual monosaccharide appear as attractive drug targets. However, although the importance of Galf in bacterial survival or pathogenesis is established, its role in eukaryotic pathogens is still undefined. Recently, we reported the identification and characterization of the first eukaryotic UDP-galactopyranose mutases. This enzyme holds a central role in Galf metabolism by providing UDP-Galf to all galactofuranosyltransferases. In this work, the therapeutical potential of Galf metabolism in Leishmania major was hence evaluated by targeted replacement of the GLF gene encoding UDP-galactopyranose mutase. In L. major, Galf is present in the membrane anchor of the lipophosphoglycan (LPG) and in glycoinositolphospholipids. Accordingly, the generated glf- mutant is deficient in LPG backbone and expresses truncated glycoinositolphospholipids. These structural changes do not influence the in vitro growth of the parasite but lead to an attenuation of virulence comparable with that observed with a mutant exclusively deficient in LPG. Considering the high incidence of galactofuranose (Galf) in pathogens and its absence from higher eukaryotes, the enzymes involved in the biosynthesis of this unusual monosaccharide appear as attractive drug targets. However, although the importance of Galf in bacterial survival or pathogenesis is established, its role in eukaryotic pathogens is still undefined. Recently, we reported the identification and characterization of the first eukaryotic UDP-galactopyranose mutases. This enzyme holds a central role in Galf metabolism by providing UDP-Galf to all galactofuranosyltransferases. In this work, the therapeutical potential of Galf metabolism in Leishmania major was hence evaluated by targeted replacement of the GLF gene encoding UDP-galactopyranose mutase. In L. major, Galf is present in the membrane anchor of the lipophosphoglycan (LPG) and in glycoinositolphospholipids. Accordingly, the generated glf- mutant is deficient in LPG backbone and expresses truncated glycoinositolphospholipids. These structural changes do not influence the in vitro growth of the parasite but lead to an attenuation of virulence comparable with that observed with a mutant exclusively deficient in LPG. Protozoan parasites of the genus Leishmania are the etiologic agents of leishmaniases, a widespread group of diseases that affect millions of people. These diseases encountered in tropical and subtropical areas of the world range from self-healing cutaneous leishmaniasis to lethal visceral leishmaniasis. Throughout the world, pentavalent antimonials have been the treatment of choice for more than 50 years. However, increasing drug resistance as well as the high cost and toxicity of these drugs considerably limit their use. Nowadays, other medications such as amphotericin B, liposomal amphotericin B, pentamidine, or miltefosine offer an alternative for treatment, but like antimonials they are toxic and/or expensive. In addition, the emergence of resistant strains, which already seriously compromise the efficacy of pentamidine, is also expected with the latest antileishmanial drug miltefosine because of its long half-life (1Croft S.L. Seifert K. Yardley V. Indian J. Med. Res. 2006; 123: 399-410PubMed Google Scholar). Efforts to develop new effective treatments have thus to be pursued. The promastigote form of Leishmania parasites transmitted to human and other mammalian reservoirs by a sandfly bite is coated by molecules of the glycosylphosphatidylinositol (GPI) 2The abbreviations used are: GPI, glycosylphosphatidylinositol; PI, phosphatidylinositol; Galf, galactofuranose; Galp, galactopyranose; GIPL, glycoinositolphospholipid; LPG, lipophosphoglycan; PPG, proteophosphoglycan; UGM, UDP-galactopyranose mutase; SAP, secreted acid phosphatase; GFP, green fluorescent protein; PBS, phosphate-buffered saline. family comprising lipophosphoglycan (LPG), GPI-anchored proteins such as membrane proteophosphoglycans (mPPG) or the metalloprotease gp63 and a heterogeneous group of glycoinositolphospholipids (GIPLs) (2McConville M.J. Ferguson M.A. Biochem. J. 1993; 294: 305-324Crossref PubMed Scopus (806) Google Scholar). Once in the mammalian host, the promastigotes enter hematopoietic cells of the monocyte/macrophage lineage by phagocytosis (3van Zandbergen G. Klinger M. Mueller A. Dannenberg S. Gebert A. Solbach W. Laskay T. J. Immunol. 2004; 173: 6521-6525Crossref PubMed Scopus (340) Google Scholar) and differentiate into amastigotes responsible for disease propagation. This latter form is covered by a thinner glycocalyx mainly composed of GIPLs and glycosphingolipids acquired from the host (4McConville M.J. Blackwell J.M. J. Biol. Chem. 1991; 266: 15170-15179Abstract Full Text PDF PubMed Google Scholar, 5Winter G. Fuchs M. McConville M.J. Stierhof Y.D. Overath P. J. Cell Sci. 1994; 107: 2471-2482Crossref PubMed Google Scholar) because both LPG and GPI-anchored proteins are strongly down-regulated at this stage. As the major macromolecule present on the promastigote, LPG has received much attention and its contribution to Leishmania pathogenesis has been critically defined. LPG is essential for the binding and detachment of parasites to the midgut of the insect vector and therefore for the transmission of the parasites to the mammalian host (6Sacks D.L. Modi G. Rowton E. Spath G. Epstein L. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 406-411Crossref PubMed Scopus (175) Google Scholar). It also protects the parasite from hydrolytic enzymes, oxidants, and human complement (7Spath G.F. Garraway L.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2003; 100: 9536-9541Crossref PubMed Scopus (224) Google Scholar). LPG is hence crucial for Leishmania major virulence as demonstrated using a mutant exclusively deficient in LPG obtained by targeted gene replacement of the putative galactofuranosyl-transferase LPG1 involved in the anchor synthesis (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar). Interestingly, LPG and some other related glycoconjugates are dispensable for L. mexicana virulence that seems to have evolved different mechanisms of host cell manipulation (9Garami A. Mehlert A. Ilg T. Mol. Cell. Biol. 2001; 21: 8168-8183Crossref PubMed Scopus (82) Google Scholar, 10Hilley J. Zawadzki J.L. McConville M.J. Coombs G.H. Mottram J.C. Mol. Cell. Biol. 2000; 11: 1183-1195Crossref Scopus (74) Google Scholar, 11Ilg T. EMBO J. 2000; 19: 1953-1962Crossref PubMed Scopus (112) Google Scholar, 12Ilg T. Demar M. Harbecke D. J. Biol. Chem. 2001; 276: 4988-4997Abstract Full Text Full Text PDF PubMed Scopus (80) Google Scholar, 13Turco S.J. Spath G.F. Beverley S.M. Trends Parasitol. 2001; 17: 223-226Abstract Full Text Full Text PDF PubMed Scopus (140) Google Scholar). In contrast, the role of GIPLs is still controversial. GIPLs are the predominant glycoconjugates of the intracellular amastigote stage and might thus play important roles in macrophage invasion and parasite survival within phagocytes. Although several studies support such roles (14Ilgoutz S.C. Zawadzki J.L. Ralton J.E. McConville M.J. EMBO J. 1999; 18: 2746-2755Crossref PubMed Scopus (76) Google Scholar, 15McNeely T.B. Rosen G. Londner M.V. Turco S.J. Biochem. J. 1989; 259: 601-604Crossref PubMed Scopus (116) Google Scholar, 16Mensa-Wilmot K. Garg N. McGwire B.S. Lu H.G. Zhong L. Armah D.A. LeBowitz J.H. Chang K.P. Mol. Biochem. Parasitol. 1999; 99: 103-116Crossref PubMed Scopus (30) Google Scholar, 17Proudfoot L. O'Donnell C.A. Liew F.Y. Eur. J. Immunol. 1995; 25: 745-750Crossref PubMed Scopus (112) Google Scholar, 18Tachado S.D. Gerold P. Schwarz R. Novakovic S. McConville M. Schofield L. Proc. Natl. Acad. Sci. U. S. A. 1997; 94: 4022-4027Crossref PubMed Scopus (185) Google Scholar), the decisive contribution of GIPLs in these processes was recently called into question by a L. major mutant deficient in all ether lipids including LPG and GIPLs (19Zufferey R. Allen S. Barron T. Sullivan D.R. Denny P.W. Almeida I.C. Smith D.F. Turco S.J. Ferguson M.A. Beverley S.M. J. Biol. Chem. 2003; 278: 44708-44718Abstract Full Text Full Text PDF PubMed Scopus (89) Google Scholar). Interestingly, LPG membrane anchor and GIPLs of L. major are structurally related molecules that contain notably a galactofuranose residue (Galf) (2McConville M.J. Ferguson M.A. Biochem. J. 1993; 294: 305-324Crossref PubMed Scopus (806) Google Scholar, 20McConville M.J. Homans S.W. Thomas-Oates J.E. Dell A. Bacic A. J. Biol. Chem. 1990; 265: 7385-7394Abstract Full Text PDF PubMed Google Scholar). This uncommon monosaccharide is highly immunogenic and present in the surface glycoconjugates of many pathogenic bacteria, fungi, and protozoan parasites (21Pedersen L.L. Turco S.J. Cell Mol. Life Sci. 2003; 60: 259-266Crossref PubMed Scopus (165) Google Scholar). It originates from the action of the UDP-galactopyranose mutase (UGM) that catalyzes the interconversion of UDP-galactopyranose (UDP-Galp) into UDP-galactofuranose (UDP-Galf) (22Bakker H. Kleczka B. Gerardy-Schahn R. Routier F.H. Biol. Chem. 2005; 386: 657-661Crossref PubMed Scopus (64) Google Scholar, 23Beverley S.M. Owens K.L. Showalter M. Griffith C.L. Doering T.L. Jones V.C. McNeil M.R. Eukaryot. Cell. 2005; 4: 1147-1154Crossref PubMed Scopus (122) Google Scholar, 24Koplin R. Brisson J.R. Whitfield C. J. Biol. Chem. 1997; 272: 4121-4128Abstract Full Text Full Text PDF PubMed Scopus (116) Google Scholar, 25Nassau P.M. Martin S.L. Brown R.E. Weston A. Monsey D. McNeil M.R. Duncan K. J. Bacteriol. 1996; 178: 1047-1052Crossref PubMed Google Scholar, A. R.E. P.M. Monsey D. Martin S.L. Duncan K. McNeil M.R. 1997; Full Text PDF PubMed Scopus Google Scholar). Galf is essential for the survival or virulence of pathogenic M. McNeil M. J. Bacteriol. 2001; PubMed Scopus Google Scholar) but is from higher eukaryotes, is a drug and has been in K. V. H. S.W. D.A. Whitfield C. McNeil M.R. J.H. J. Mol. Biol. 2005; PubMed Scopus Google Scholar, L.L. Chem. Biol. 2006; Full Text Full Text PDF PubMed Scopus Google Scholar, D.A. McNeil M.R. Whitfield C. J.H. Biol. 2001; PubMed Scopus Google Scholar, S.W. S. D.A. Whitfield C. J.H. 2003; PubMed Scopus Google Scholar, M. J.H. L.L. J. Chem. 2004; PubMed Scopus Google Scholar, M. L.L. Mol. Biol. 2004; 11: PubMed Scopus Google Scholar). Recently, we and and the first eukaryotic from the human pathogens L. major, and (22Bakker H. Kleczka B. Gerardy-Schahn R. Routier F.H. Biol. Chem. 2005; 386: 657-661Crossref PubMed Scopus (64) Google Scholar, 23Beverley S.M. Owens K.L. Showalter M. Griffith C.L. Doering T.L. Jones V.C. McNeil M.R. Eukaryot. Cell. 2005; 4: 1147-1154Crossref PubMed Scopus (122) Google Scholar). Interestingly, the of the GLF gene that Galf is more widespread in than its role in is still undefined. In this we the role of Galf in L. major pathogenesis by targeted replacement of of is expected to affect not the synthesis of the virulence LPG but also that of the L. major at putative to LPG1 are still K. Barron T. Turco S.J. Beverley S.M. Mol. Biochem. Parasitol. 2004; PubMed Scopus Google Scholar). a central enzyme of Galf metabolism such as than have a more on Leishmania and from M. Beverley E. Proc. Natl. Acad. Sci. U. S. A. 1993; PubMed Scopus Google Scholar) was by W. of L. major at in with and 50 As B, and at and of was on of L. major of the of GLF and a of the by from L. major using the and and the and used for into in The was by the resistance gene from with and and in by in as by the The was used to the from using the and to are The and of the resistance of GLF in the and with and and the from and by The mutant was first obtained by of of in L. major as J.H. D. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 1990; PubMed Scopus Google Scholar). with the in to as glf- GLF was in several of the glf- mutant by with of The latter was by of GLF with the and and in the and of Schwarz Turco S.J. Beverley S.M. Mol. Biochem. Parasitol. 1996; PubMed Scopus Google Scholar). of the are of L. major an UGM, the the was using the and and in the into the of Schwarz Turco S.J. Beverley S.M. Mol. Biochem. Parasitol. 1996; PubMed Scopus Google Scholar). of the in L. major and in the glf- to and in in for at in was to the and by was used as cell of promastigotes by and to The D. PubMed Scopus Google Scholar) and a to used at a of and was used at a of of a was with an by glf- and promastigotes in PBS, and in for at and with 50 for and with and in The and the used at a of and was used at a of of a with for in the in and by and of from cell of promastigotes by and by with at a L. mexicana secreted acid was in the different cell by with and with the was by with and using the and of of parasites with was by of in of and at for The obtained from a in of and to a with of and of the with of GIPLs in of a and in of of of and of on a a on a in the The was used at a potential of first and for than in promastigotes into the of and by the of the with a and to the in the parasites by Zandbergen G. A. A. S. R. Klinger M. A. C. M. D. Solbach W. Laskay T. Proc. Natl. Acad. Sci. U. S. A. 2006; PubMed Scopus Google Scholar). from into and LPG was by as L. major in the by GLF was (22Bakker H. Kleczka B. Gerardy-Schahn R. Routier F.H. Biol. Chem. 2005; 386: 657-661Crossref PubMed Scopus (64) Google Scholar, 23Beverley S.M. Owens K.L. Showalter M. Griffith C.L. Doering T.L. Jones V.C. McNeil M.R. Eukaryot. Cell. 2005; 4: 1147-1154Crossref PubMed Scopus (122) Google Scholar). is in the of of membrane or that this is However, is in and other for the of proteins involved in interconversion C.L. L. Doering T.L. J. Biol. Chem. 2004; Full Text Full Text PDF PubMed Scopus Google Scholar, M. PubMed Scopus Google Scholar, J.R. A. Ferguson M.A. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar). The of was thus using of in L. major and glf- mutant that the is the cell within L. major the of The is with the absence of or in acid As a LPG that is from the glf- mutant was that the is not of L. major that GLF is a gene not which was by targeted gene L. major was recently L. G. M. E. M.A. E. R. A. P. N. C. A. Beverley S.M. G. K. G. M. E. L. C. A. J. A. G. N. A. Fuchs M. C. A. A. D. C. H. D. S. A. M. N. L. A. T. M. D. K. S. Mottram J.C. S. H. S. H. K. S. M. C. B. M.A. E. R. M. J. J. L. J.C. S. D. M. J. K. A. S. H. D. R. S. V. C. G. R. T. H. J. S. W. Smith D.F. Blackwell J.M. B. 2005; PubMed Scopus Google Scholar) and a of GLF present on is that gene in Leishmania is by present in the and the by to a replacement of GLF by the resistance and thus of these of gene replacement to a mutant because Leishmania has a of from and targeted parasites with a GLF to the of observed in and In contrast, was observed in the glf- of both GLF of the and at the was by an with a in the GLF the used for using for and of the not glf- and in as well as the As a for several of glf- mutant with an GLF In all and thus from a glf- and mutant be The glf- of in L. major and was first by using the This the present in L. major LPG and and strongly the cell surface of and In contrast, fluorescent was at the cell surface of the glf- the absence of of LPG was by of cell with of LPG is observed with L. major but not with the glf- mutant As of LPG synthesis of in and from and cell by and by with of in the obtained from L. major and cell In contrast, in the in with L. major not These the of in L. major and are with the absence of of glf- parasites with L. mexicana secreted acid was in L. major and the glf- with the and by with and As was in cell with and not in cell by the glf- mutant more with molecules by that the of is in this cell L. major glf- GIPLs and LPG from and glf- and by in the L. major different GIPLs of and as and to the of that they contain M.J. Homans S.W. Thomas-Oates J.E. Dell A. Bacic A. J. Biol. Chem. 1990; 265: 7385-7394Abstract Full Text PDF PubMed Google Scholar). of in the be observed in the of L. major GIPLs The at and and with and or the at and to with and or These by and are in with the in L. major M.J. Homans S.W. Thomas-Oates J.E. Dell A. Bacic A. J. Biol. Chem. 1990; 265: 7385-7394Abstract Full Text PDF PubMed Google Scholar, P. C.L. Ferguson M.A. McConville M.J. Biochem. J. 1994; PubMed Scopus Google Scholar). a with is also present in the The latter was not reported in L. major but seems to be present in L. major (19Zufferey R. Allen S. Barron T. Sullivan D.R. Denny P.W. Almeida I.C. Smith D.F. Turco S.J. Ferguson M.A. Beverley S.M. J. Biol. Chem. 2003; 278: 44708-44718Abstract Full Text Full Text PDF PubMed Scopus (89) Google Scholar). The of GIPLs from mutant was to the not In contrast, of glf- mutant the of truncated GIPLs As expected the biosynthesis of GIPLs seems to the of the to GIPLs the as The as in GIPLs to the of different at and As a of UDP-Galf LPG biosynthesis is also to the of a LPG anchor with the The is therefore by an at to a of this LPG anchor with a The of a is by the of in this The at to the a of truncated LPG anchor with a be observed at by Interestingly, molecules with a and a or at and are not in the glf- although these are the lipids in at and at and and at and As of the but not of the has been J.E. McConville M.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar), this that these molecules be LPG biosynthesis by and The glf- of GPI-anchored synthesis of the GPI-anchored metalloprotease gp63 was by and with the L. was used to of As of GLF not affect the or of gp63 not for of in virulence of the glf- mutant was by of The in are the of with or at the of to In contrast, disease was to in with the glf- In both and the with parasite glf- parasites from not LPG, thus the of In different enzymes involved in Galf metabolism have been in and the crucial role of Galf for survival or virulence of several pathogenic (21Pedersen L.L. Turco S.J. Cell Mol. Life Sci. 2003; 60: 259-266Crossref PubMed Scopus (165) Google Scholar). In contrast, the role of Galf in is still undefined. In this work, the role of Galf for L. major virulence was by targeted of the GLF gene encoding and recently and this central enzyme in Galf biosynthesis that into UDP-Galf (22Bakker H. Kleczka B. Gerardy-Schahn R. Routier F.H. Biol. Chem. 2005; 386: 657-661Crossref PubMed Scopus (64) Google Scholar, 23Beverley S.M. Owens K.L. Showalter M. Griffith C.L. Doering T.L. Jones V.C. McNeil M.R. Eukaryot. Cell. 2005; 4: 1147-1154Crossref PubMed Scopus (122) Google Scholar). L. major was in the as from the of of membrane or Interestingly, the of has been in the J.R. A. Ferguson M.A. J. Biol. Chem. 2005; Full Text Full Text PDF PubMed Scopus Google Scholar) and several other enzymes involved in are to be in this Mol. Biochem. Parasitol. 2006; PubMed Scopus Google Scholar). The of thus the of a from the into the and of a UDP-Galf because the LPG1 is in this Schwarz Turco S.J. Beverley S.M. Mol. Biochem. Parasitol. 1996; PubMed Scopus Google Scholar). The latter is expected to be from In L. major Galf is present in the anchor of the cell surface LPG as in other Leishmania and in the GIPLs (2McConville M.J. Ferguson M.A. Biochem. J. 1993; 294: 305-324Crossref PubMed Scopus (806) Google Scholar). Accordingly, targeted replacement of GLF in the absence of LPG backbone and the of truncated GIPLs to the present in L. of secreted acid was in the glf- influence on was also observed in the mutant that both LPG and GIPLs (19Zufferey R. Allen S. Barron T. Sullivan D.R. Denny P.W. Almeida I.C. Smith D.F. Turco S.J. Ferguson M.A. Beverley S.M. J. Biol. Chem. 2003; 278: 44708-44718Abstract Full Text Full Text PDF PubMed Scopus (89) Google Scholar) and be as an of LPG synthesis because the and LPG are However, such of was not reported in L. major and L. mexicana that are exclusively deficient in LPG (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar, 11Ilg T. EMBO J. 2000; 19: 1953-1962Crossref PubMed Scopus (112) Google Scholar) and its to be The structural changes from absence not affect the or growth of Leishmania in This was because other in or more surface glycoconjugates are and do not growth in In contrast, of GLF the growth of the and H. Routier and W. in the influence of Galf metabolism on growth has also been observed in P.M. Martin S.L. Brown R.E. Weston A. Monsey D. McNeil M.R. Duncan K. J. Bacteriol. 1996; 178: 1047-1052Crossref PubMed Google Scholar, M. McNeil M. J. Bacteriol. 2001; PubMed Scopus Google Scholar). L. major GIPLs that be from the LPG anchor by the absence of residue on the and by their LPG is exclusively a with long or GIPLs contain with or (2McConville M.J. Ferguson M.A. Biochem. J. 1993; 294: 305-324Crossref PubMed Scopus (806) Google Scholar). that the of GIPLs by L. major are considerably in the glf- In the absence of Galf addition, these molecules to be and by the of a residue to the LPG anchor that in the These strongly that and LPG from a and thus a L. P. Ferguson M.A. McConville M.J. Biochem. J. 1995; PubMed Scopus Google Scholar). In L. mexicana that exclusively GIPLs with a GIPLs and LPG anchor are to be the of J.E. McConville M.J. J. Biol. Chem. Full Text Full Text PDF PubMed Scopus Google Scholar). of the of on and on the other might also in L. The of in L. major amastigotes in which the LPG is strongly down-regulated P. A. Ferguson M.A. McConville M.J. Biochem. J. 1993; PubMed Scopus Google Scholar) and the of in LPG and GIPLs biosynthesis (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar) be with this is still different are involved in the biosynthesis of and LPG (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar) and thus the of these molecules the of the of LPG with a residue and a in the glf- mutant that the enzymes involved in these do not the of As are not by these enzymes because of their or because of of the of the putative LPG1 that in the of LPG, was that LPG is a virulence in L. major (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar). the of L. major glf- virulence observed in a was The role of on the other is more LPG and the major surface they are in both promastigote and amastigote The molecules present at the surface of amastigote are to of the parasite membrane P. A. Ferguson M.A. McConville M.J. Biochem. J. 1993; PubMed Scopus Google Scholar, McConville M. Homans S.W. 1994; PubMed Scopus Google Scholar) and play a the Galf residue present in was to in the of the parasites with macrophage and their E. PubMed Scopus Google Scholar). Galf be at the of GIPLs and thus McConville M. Homans S.W. 1994; PubMed Scopus Google Scholar). the mutant that is deficient in all ether including LPG and GIPLs the importance of GIPLs for Leishmania pathogenesis (19Zufferey R. Allen S. Barron T. Sullivan D.R. Denny P.W. Almeida I.C. Smith D.F. Turco S.J. Ferguson M.A. Beverley S.M. J. Biol. Chem. 2003; 278: 44708-44718Abstract Full Text Full Text PDF PubMed Scopus (89) Google Scholar). In with this we that the of Galf by the absence of LPG backbone and of the GIPLs in a of disease comparable with that obtained LPG is deficient (8Spath G.F. Epstein L. Leader B. Singer S.M. Avila H.A. Turco S.J. Beverley S.M. Proc. Natl. Acad. Sci. U. S. A. 2000; 97: 9258-9263Crossref PubMed Scopus (252) Google Scholar). the of GIPLs on the virulence of Leishmania Once has been established, disease in with Leishmania or glf- is that Galf is not essential for the or the survival of amastigotes into these phagocytes. observed might thus be to the of LPG. Galf to Leishmania major M. Beverley for the of W. for providing gp63 and and for with the with Solbach and Laskay

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