Role of 67 kDa cell surface laminin binding protein of Leishmania donovani in pathogenesis.
The role that interaction with laminin may play in Leishmania donovani infection was investigated. Binding of (125)I-radiolabeled laminin, in a liquid-phase assay, by the parasite was rapid, saturable, specific, reversible, and of high affinity. Using a Western blotting procedure, a 67 kDa laminin-binding protein (LBP) was identified from the membrane of both the promastigote and amastigote forms of L. donovani. Subsequently, the protein was purified by affinity chromatography. Immunofluorescence with a polyclonal antibody against LBP as well as flow cytometric analysis demonstrated its presence at the parasite surface. After stimulation with phorbol-12-myristate-13-acetate (PMA), U937 cells exhibited the ability to adhere to laminin and LBP specifically inhibited this adhesion. The reduced parasite adhesion after tunicamycin treatment suggested the importance of sugar residues in cell adhesion. Although co-administration of either laminin or LBP or anti LBP antibody reduced parasite virulence, resulting in a lower level of infection in the BALB/c mouse model, an in vitro macrophage culture-enhanced level of infection was observed in the case of laminin-coated parasites. The results collectively suggest a role for LBP in the interaction of the parasite with extracellular matrix elements, which may constitute a basis for the homing of the parasite to its physiological address.
Bandyopadhyay
K
K
Molecular Cell Biology Laboratory, Indian Institute of Chemical Biology, Raja S.C. Mullick Road, Jadavpur, Calcutta 700 032, India.
Karmakar
S
S
Ghosh
A
A
Das
P K
PK
D000818:Animals; D000906:Antibodies / Q000494:pharmacology / Q000627:therapeutic use; D000954:Antigens, Surface / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D015153:Blotting, Western; D002448:Cell Adhesion / Q000187:drug effects; D005434:Flow Cytometry; D006801:Humans; D007797:Laminin / Q000378:metabolism* / Q000494:pharmacology / Q000627:therapeutic use; D007893:Leishmania donovani / Q000737:chemistry / Q000187:drug effects / Q000378:metabolism* / Q000472:pathogenicity*; D007898:Leishmaniasis, Visceral / Q000188:drug therapy / Q000378:metabolism / Q000469:parasitology / Q000473:pathology; D008565:Membrane Proteins / Q000037:antagonists & inhibitors / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism*; D051379:Mice; D008807:Mice, Inbred BALB C; D008856:Microscopy, Fluorescence; D008970:Molecular Weight; D011485:Protein Binding / Q000187:drug effects; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D013997:Time Factors; D014415:Tunicamycin / Q000494:pharmacology; D020298:U937 Cells; D014774:Virulence / Q000187:drug effects
D000906:Antibodies; D000954:Antigens, Surface; D007797:Laminin; D008565:Membrane Proteins; D014415:Tunicamycin; D013755:Tetradecanoylphorbol Acetate