Characterization of integral membrane proteins of Leishmania major by Triton X-114 fractionation and analysis of vaccination effects in mice.
The total integral membrane proteins of promastigotes of Leishmania major were extracted by using the Triton X-114 phase separation technique and were characterized by immunoprecipitation, Western blotting (immunoblotting), and lectin chromatography. Of the 40 or more proteins which partitioned into the detergent phase, only about 10 proteins could be surface radioiodinated on live promastigotes, suggesting their surface orientation. The abundance of the gp58-63 antigen varied markedly between two strains of L. major. Sera from patients with visceral leishmaniasis caused by Leishmania donovani chagasi recognized the gp58-63 complex and an additional Mr-42,000 polypeptide shared between L. major and L. donovani chagasi. A subpopulation of six surface proteins, including the abundant gp58-63 antigen and a group of proteins of Mr 81,000 to 105,000, were glycoproteins recognized by antiserum to wheat germ agglutinin- or concanavalin A-binding proteins. The membrane proteins of the LRC-L119 isolate of L. major could successfully vaccinate genetically susceptible mice, thus opening the way for a molecularly defined subunit vaccine composed of glycolipid and membrane protein antigens.
Murray
P J
PJ
Walter and Eliza Hall Institute of Medical Research, Royal Melbourne Hospital, Victoria, Australia.
Spithill
T W
TW
Handman
E
E
D000818:Animals; D000953:Antigens, Protozoan / Q000008:administration & dosage / Q000302:isolation & purification; D005591:Chemical Fractionation / Q000379:methods; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D015151:Immunoblotting; D007895:Leishmania tropica / Q000032:analysis* / Q000276:immunology; D007896:Leishmaniasis / Q000517:prevention & control; D008565:Membrane Proteins / Q000008:administration & dosage / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D017830:Octoxynol; D011092:Polyethylene Glycols; D011233:Precipitin Tests; D013501:Surface-Active Agents; D014612:Vaccines / Q000008:administration & dosage*
D000953:Antigens, Protozoan; D003902:Detergents; D008565:Membrane Proteins; D013501:Surface-Active Agents; D014612:Vaccines; D011092:Polyethylene Glycols; D017830:Octoxynol; C010615:Nonidet P-40