The Journal of Immunology, Vol 141, Issue 1 265-272, Copyright © 1988 by American Association of Immunologists

ARTICLES

The major concanavalin A-binding surface glycoprotein of Leishmania donovani chagasi promastigotes is involved in attachment to human macrophages

ME Wilson and KK Hardin
Department of Internal Medicine, Veterans Administration Medical Center, Iowa City.

Leishmania donovani, the protozoan causing visceral leishmaniasis, is an obligate intracellular parasite of mammalian macrophages. Considerable evidence has suggested that the ingestion of L. donovani promastigotes by macrophages occurs via receptors on the surface of the phagocyte. During this study, a glycoconjugate that may be involved in the receptor-mediated ingestion of L. donovani chagasi promastigotes was isolated from the parasite membrane. Octyl glucoside-soluble extracts of promastigote membranes contained a predominant doublet migrating at 60 kDa, seen by SDS-PAGE. The 60-kDa molecule was the major externally disposed promastigote surface protein labeled by 125I, and it was the major Con A-binding protein on L. donovani chagasi, as determined by Con A binding to parasite proteins transferred to nitrocellulose. Attachment of promastigotes to human monocyte-derived macrophages was inhibited by varying concentrations of the membrane extract containing both proteins, and adsorption of extracts on Con A- Sepharose resulted in both removal of the 60,000 Mr glycoprotein and loss of the ability of extracts to inhibit promastigote attachment to human macrophages. After further purification of the 60-kDa glycoprotein by gel filtration, its inhibitory activity increased 45- fold over the unpurified membrane extract. Examination of Con A blots of stationary phase promastigotes isolated from an infected hamster revealed a marked loss in the major Con A-binding glycoprotein over 4 mo in in vitro culture after isolation from the rodent host, corresponding to a loss in infectivity of the promastigotes for hamsters. The results suggest that the major Con A-binding surface glycoprotein from L. donovani chagasi promastigotes is important in attachment to human macrophages, and may be a factor in parasite virulence for a mammalian host.

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