In silico characterisation of putative Plasmodium falciparum vaccine candidates in African malaria populations.

Genetic diversity of surface exposed and stage specific Plasmodium falciparum immunogenic proteins pose a major roadblock to developing an effective malaria vaccine with broad and long-lasting immunity. We conducted a prospective genetic analysis of candidate antigens (msp1, ama1, rh5, eba175, glurp, celtos, csp, lsa3, Pfsea, trap, conserved chrom3, hyp9, hyp10, phistb, surfin8.2, and surfin14.1) for malaria vaccine development on 2375 P. falciparum sequences from 16 African countries. We described signatures of balancing selection inferred from positive values of Tajima's D for all antigens across all populations except for glurp. This could be as a result of immune selection on these antigens as positive Tajima's D values mapped to regions with putative immune epitopes. A less diverse phistb antigen was characterised with a transmembrane domain, glycophosphatidyl anchors between the N and C- terminals, and surface epitopes that could be targets of immune recognition. This study demonstrates the value of population genetic and immunoinformatic analysis for identifying and characterising new putative vaccine candidates towards improving strain transcending immunity, and vaccine efficacy across all endemic populations.

Genetic diversity of Plasmodium falciparum in Bolifamba, on the slopes of Mount Cameroon: influence of MSP1 allelic variants on symptomatic malaria and anaemia.

The influence of the genetic diversity of Plasmodium falciparum infection on the clinical presentation of human malaria was investigated in rural Bolifamba, Cameroon. Parasite DNA was obtained from the blood of 208 children (aged 1-15 years) with malarial infection. The prevalences of anaemia and symptomatic and asymptomatic malaria among these children were 57.7%, 51.4% and 48.6%, respectively. The frequencies of parasites carrying each of the block-2 allelic variants (MAD20, K1 and RO33) of merozoite surface protein 1 (MSP1) were compared among the symptomatic and asymptomatic cases of malaria, the anaemic and non-anaemic subjects, and in various age groupings. Although all three allelic variants were found in Bolifamba, 32.7% of the children investigated were co-infected with parasites carrying the RO33 and K1 variants. There was a significant difference in the prevalence of each MSP1 allelic variant both between age-groups and between the various categories of anaemia considered (P<0.0001 for each), with the highest number of alleles occurring in the children with severe anaemia. The combination of RO33/K1 co-infection and anaemia was detected in most (57.7%) of the children aged 3-<6 years. The RO33/K1 co-infection was also strongly associated with both fever and high levels of parasitaemia (P<0.0001 for each). Although the children of Bolifamba are exposed to all three allelic variants of MSP1, which occur either singly or in varying combinations in the infected children, RO33/K1 co-infections are particularly associated with fever and this association appears independent of age and parasite density. The preliminary data presented here should facilitate the design of future research towards the development and testing of malaria candidate vaccines in the study area.

The FAR protein family of the nematode Caenorhabditis elegans. Differential lipid binding properties, structural characteristics, and developmental regulation.

Parasitic nematodes of humans and plants secrete a structurally novel type of fatty acid- and retinol-binding protein, FAR, into the tissues they occupy. These proteins may interfere with intercellular lipid signaling to manipulate the defense reactions of the host or acquire essential lipids for the parasites. The genome of the nematode Caenorhabditis elegans encodes eight FAR-like proteins (Ce-FAR-1 to -8). These fall into three discrete groups as indicated by phylogenetic sequence comparisons and intron positions, the proteins from parasitic nematodes falling into group A. Recombinant Ce-FAR-1 to -7 were produced in Escherichia coli and tested for lipid binding in fluorescence-based assays. Ce-FAR-1 to -6 bound DAUDA (11-((5-dimethylaminonaphthalene-1-sulfonyl)amino)undecanoic acid), cis-parinaric acid, and retinol with dissociation constants in the micromolar range, whereas Ce-FAR-7 bound the latter two lipids relatively poorly. Each protein produced a characteristic shift in peak fluorescence emission of DAUDA, and one (Ce-FAR-5) produced a shift greater than has been observed previously for any lipid-binding protein. Selected Ce-FAR proteins were analyzed by circular dichroism (CD) and differential scanning calorimetry, were found to be helix-rich, and exhibited high thermal stability (transition midpoint, 82.7 degrees C). CD and secondary structure predictions, however, both indicated that Ce-FAR-7 possesses substantially less helix than the other FAR proteins. The genes encoding the Ce-FAR proteins were found to be transcribed differentially through the life cycle of C. elegans, such that Ce-far-4 was transcribed at highest levels in the fourth larval stage, and Ce-far-3 and -7 predominated in males.