Molecular and immunological characterisation of the glucose regulated protein 78 of Leishmania donovani(1).
To identify novel potential Leishmania vaccine antigens, antibodies from patients with visceral leishmaniasis (VL) were used to isolate clones from a cDNA expression library of L. donovani amastigotes. Glucose Regulated Protein (GRP78), a member of the 70 kDa heat-shock protein family was identified and characterised. The GRP78 gene was localised to chromosome 15 in L. donovani, L. major, and L. mexicana by pulse-field gel electrophoresis. The Leishmania GRP78 protein contain a carboxy-terminal endoplasmic reticulum retention signal sequence (MDDL) as does the Trypanosoma cruzi GRP78. Immunofluorescence using antibodies to the recombinant DNA-derived GRP78 protein showed staining localised to reticular material throughout the cytoplasm and in the perinuclear region of promastigotes, suggesting that the protein is localised in the endoplasmic reticulum. The protective efficacy of GRP78 was assessed in mice vaccine experiments. A GRP78 DNA vaccine primed for an immune response that protected C57Bl/6 and C3H/He mice against infection with L. major. Similarly vaccination with a recombinant form of GRP78 purified from Escherichia coli and administered with Freund's as adjuvant induced protective immunity in C57Bl/6 mice.
Jensen
A T
AT
Centre for Medical Parasitology, Institue for Medical Microbiology and Immunology, University of Copenhagen, The Panum Institute, Denmark. atjr@biobase.dk
Curtis
J
J
Montgomery
J
J
Handman
E
E
Theander
T G
TG
D000595:Amino Acid Sequence; D000818:Animals; D000953:Antigens, Protozoan / Q000276:immunology; D001483:Base Sequence; D015152:Blotting, Northern; D003001:Cloning, Molecular; D004195:Disease Models, Animal; D016521:Electrophoresis, Gel, Pulsed-Field; D000091342:Endoplasmic Reticulum Chaperone BiP; D015723:Gene Library; D017125:Genes, Protozoan; D018840:HSP70 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism; D006801:Humans; D007893:Leishmania donovani / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D007896:Leishmaniasis / Q000276:immunology; D051379:Mice; D008809:Mice, Inbred C3H; D008810:Mice, Inbred C57BL; D018613:Microscopy, Confocal; D008969:Molecular Sequence Data; D015800:Protozoan Proteins / Q000235:genetics / Q000276:immunology / Q000378:metabolism*; D016052:Protozoan Vaccines / Q000008:administration & dosage / Q000276:immunology*; D014611:Vaccination; D019444:Vaccines, DNA / Q000008:administration & dosage
D000953:Antigens, Protozoan; D000091342:Endoplasmic Reticulum Chaperone BiP; D018840:HSP70 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D015800:Protozoan Proteins; D016052:Protozoan Vaccines; D019444:Vaccines, DNA; C436924:glucose regulated protein 78, Leishmania donovani