Analysis of human antibodies to erythrocyte binding antigen 175 of Plasmodium falciparum.

Invasion of human erythrocytes by Plasmodium falciparum merozoites is a multistep process. For many strains of the parasite, part of this process requires that the erythrocyte binding antigen 175 (EBA-175) of the merozoite binds to sialic acid residues of glycophorin A on the erythrocyte surface, a receptor-ligand interaction which represents a potential target for inhibition by antibodies. This study characterizes the reactivity of naturally acquired human antibodies with four recombinant proteins representing parts of EBA-175 (region II, regions III to V, and the dimorphic C and F segment region) in populations in which the organism is endemic. Serum immunoglobulin G (IgG) recognizing the recombinant proteins is predominantly of the IgG1 and IgG3 subclasses, and its prevalence increases with age. In a large population study in The Gambia, serum positivity for IgG or IgG1 and IgG3 subclass antibodies to each of the EBA-175 recombinant antigens was not significantly associated with subsequent protection from clinical malaria. However, there was a trend indicating that individuals with high levels of IgG to region II may have some protection.

Purification and characterisation of an extracellularly released protease of Trypanosoma brucei.

Thrombocytopaenia, or platelet aggregation, is a serious complication of African trypanosomiasis. The biochemical basis is not clearly known. Proteases are known potent inducers of blood coagulation and platelet aggregation, and unknown factors released by Trypanosoma brucei have been shown to induce platelet aggregation. In attempts to define the biochemical mechanisms involved in thrombocytopaenia we purified and characterised a major proteolytic enzyme released extracellularly by T. brucei. Actively motile trypanosomes released proteins into the medium (phosphate saline/glucose, pH 8.0) in which the organisms were incubated in vitro. The M(r) of the released polypeptides ranged from 15 to > 200 kDa, amongst which are proteases. One of the major protein bands, a 250 kDa protease, was purified to homogeneity by ammonium sulfate precipitation followed by diethylaminoethyl (DEAE)-cellulose chromatography and Sephacryl S-300 gel filtration. The protease migrated as a single band of 63 kDa upon electrophoresis in both denaturing and non-denaturing gel co-polymerised with gelatin. The enzyme was strongly active against Z-ARR-AFC peptide substrate, with a pH optimum of 7.0. The proteolytic activity was enhanced by dithiothreitol and inhibited by E-64, leupeptin, TPCK and antipain. The released proteolytic enzyme is putatively identified as a cathepsin B-like cysteine protease.