Characterization of lipophosphoglycan from a ricin-resistant mutant of Leishmania major.

Abstract

One of the virulence factors of the protozoan parasite Leishmania major is the surface glycoconjugate, lipophosphoglycan (LPG). A Ricin-resistant mutant of L.major was generated and characterised with respect to its virulence in mice and the structure and expression of LPG. The LPG from this mutant (1F6-B5) retained the tripartite structure of wild-type LPG, comprising a glycosylphosphatidylinositol (GPI) anchor linked to a phosphorylated disaccharide backbone terminating in a nonreducing neutral oligosaccharide cap. The structure of the GPI anchor and the major capping oligosaccharide were identical to wild-type LPG. However, there were variations in the number of phosphorylated repeats (PO4-6Gal(beta 1-4)Man(alpha 1-) comprising the backbone of LPG, although the degree of substitution with side branches (approx. 95%) was similar to that of wild-type LPG. Thus, the mutant LPG was shorter in length having, on average, 15 repeat units per molecule compared with 30 in the wild-type LPG. The mutant LPG contained both arabinose (Ara(beta 1-2)[Gal(beta 1-3)-]1,2) and galactose ([Gal(beta 1-3)-]1-8) capped side branches linked to the backbone. In contrast to wild-type LPG, the number of arabinose-capped side chains was significantly reduced, and a new population of galactose-capped (Gal(beta 1-3)]5-8) side branches was present. The level of LPG expression in mutant parasites was approximately one-tenth of the wild-type parasite. The mutant parasites were avirulent in mice. Over a period of 18 months, they did not cause lesions and organisms could not be isolated from the draining lymph nodes.