Cellular responses to modified Plasmodium falciparum MSP119 antigens in individuals previously exposed to natural malaria infection

<p>Abstract</p> <p>Background</p> <p>MSP1 processing-inhibitory antibodies bind to epitopes on the 19 kDa C-terminal region of the <it>Plasmodium falciparum </it>merozoite surface protein 1 (MSP1₁₉), inhibiting erythrocyte invasion. Blocking antibodies also bind to this antigen but prevent inhibitory antibodies binding, allowing invasion to proceed. Recombinant MSP1₁₉had been modified previously to allow inhibitory but not blocking antibodies to continue to bind. Immunization with these modified proteins, therefore, has the potential to induce more effective protective antibodies. However, it was unclear whether the modification of MSP1₁₉would affect critical T-cell responses to epitopes in this antigen.</p> <p>Methods</p> <p>The cellular responses to wild-type MSP1₁₉and a panel of modified MSP1₁₉antigens were measured using an <it>in-vitro </it>assay for two groups of individuals: the first were malaria-naïve and the second had been naturally exposed to <it>Plasmodium falciparum </it>infection. The cellular responses to the modified proteins were examined using cells from malaria-exposed infants and adults.</p> <p>Results</p> <p>Interestingly, stimulation indices (SI) for responses induced by some of the modified proteins were at least two-fold higher than those elicited by the wild-type MSP1₁₉. A protein with four amino acid substitutions (Glu27→Tyr, Leu31→Arg, Tyr34→Ser and Glu43→Leu) had the highest stimulation index (SI up to 360) and induced large responses in 64% of the samples that had significant cellular responses to the modified proteins.</p> <p>Conclusion</p> <p>This study suggests that specific MSP1₁₉variants that have been engineered to improve their antigenicity for inhibitory antibodies, retain T-cell epitopes and the ability to induce cellular responses. These proteins are candidates for the development of MSP1-based malaria vaccines.</p

Land use/land cover change, physico-chemical parameters and freshwater snails in Yewa North, Southwestern Nigeria.

The management of ecosystem has been a major contributor to the control of diseases that are transmitted by snail intermediate hosts. The ability of freshwater snails to self-fertilize, giving rise to thousands of hatchlings, enables them to contribute immensely to the difficulty in reducing the endemicity of some infections in the world. One of the effects of land use/land cover change (LU/LCC) is deforestation, which, in turn, leads to the creation of suitable habitats for the survival of freshwater snails. This study was aimed at studying the land use/land cover change, physico-chemical parameters of water bodies and to understand the interplay between them and freshwater snails in an environment where a new industrial plant was established. Landsat TM, 1984, Landsat ETM+ 2000 and Operational land Imager (OLI) 2014 imageries of the study area were digitally processed using ERDAS Imagine. The land use classification includes settlement, water bodies, wetlands, vegetation and exposed surface. Dissolved oxygen, water temperature, pH, total dissolved solids and conductivity were measured with multipurpose digital meters. Snail sampling was done at each site for 30 minutes along the littoral zones, using a long-handled scoop (0.2mm mesh size) net once every month for 24 months. Independent t-test was used to determine the variation between seasons, Spearman's rank correlation coefficient was used to test the relationship between physico-chemical parameters and snail species while regression was used to analyze the relationship between LU/LCC and freshwater snails. Species' richness, diversity and evenness were examined using Margalef, Shannon Weiner and Equitability indexes. Snail species recovered include: Bulinus globosus, Bulinus jousseaumei, Bulinus camerunensis, Bulinus senegalensis, Bulinus forskalii, Amerianna carinatus, Ferrissia spp., Segmentorbis augustus, Lymnaea natalensis, Melanoides tuberculata, Physa acuta, Gyraulus costulatus, Indoplanorbis exuxtus and Gibbiella species. Out of the total snails recovered, M. tuberculata (2907) was the most abundant, followed by Lymnaea natalensis (1542). The highest number of snail species was recovered from Iho River while the least number of snails was recovered from Euro River. The mean and standard deviation of physico-chemical parameters of the water bodies were DO (2.13±0.9 mg/L), pH (6.80±0.4), TDS (50.58±18.8 mg/L), Temperature (26.2±0.9°C) and Conductivity (74.00±27.5 μS/cm). There was significant positive correlation between pH and B. globosus (r = 0.439; P<0.05). Dissolved oxygen showed significant positive correlation with B. globosus (r = 0.454; P<0.05) and M. tuberculata (r = 0.687; P<0.01). There was a positive significant relationship between LULCC and B. camerunensis (p<0.05). The positive relationship between LULCC and the abundance of B. globosus, B. jousseaumei was not significant. The area covered by water bodies increased from 3.72 to 4.51 kilometers; this indicates that, more suitable habitats were being created for the multiplication of freshwater snails. We therefore conclude that, increase in areas suitable for the survival of freshwater snails could lead to an increase in water-borne diseases caused by the availability of snail intermediate hosts

Acquisition, maintenance and adaptation of invasion inhibitory antibodies against Plasmodium falciparum invasion ligands involved in immune evasion

Erythrocyte-binding antigens (EBAs) and P. falciparum reticulocyte-binding homologue proteins (PfRhs) are two important protein families that can vary in expression and utilization by P. falciparum to evade inhibitory antibodies. We evaluated antibodies at repeated time-points among individuals living in an endemic region in Nigeria over almost one year against these vaccine candidates. Antibody levels against EBA140, EBA175, EBA181, PfRh2, PfRh4, and MSP2, were measured by ELISA. We also used parasites with disrupted EBA140, EBA175 and EBA181 genes to show that all these were targets of invasion inhibitory antibodies. However, antigenic targets of inhibitory antibodies were not stable and changed substantially over time in most individuals, independent of age. Antibodies levels measured by ELISA also varied within and between individuals over time and the antibodies against EBA181, PfRh2 and MSP2 declined more rapidly in younger individuals (15 years) compared with older (>15). The breadth of high antibody responses over time was more influenced by age than by the frequency of infection. High antibody levels were associated with a more stable invasion inhibitory response, which could indicate that during the long process of formation of immunity, many changes not only in levels but also in functional responses are needed. This is an important finding in understanding natural immunity against malaria, which is essential for making an efficacious vaccine

Kaplan- Meier survival curves showing the proportion of individuals with antibody levels of >50% of the day 0 antibody level for all antigens among the different age groups and among individuals with different levels of exposure to <i>P</i>. <i>falciparum</i>.

<p>Kaplan- Meier survival curves showing the proportion of individuals with antibody levels of >50% of the day 0 antibody level for all antigens among the different age groups and among individuals with different levels of exposure to <i>P</i>. <i>falciparum</i>.</p

Relationship between invasion inhibitory antibodies to invasion pathway ligands parasitaemia and age.

<p>(A) Effects of antibodies directed against SA-dependent or SA-independent pathways on the mean parasitaemia. “<b>A</b>”, “<b>B</b>” represent individuals with only antibody responses to SA-dependent or SA-independent pathway invasion ligands, respectively. “<b>A and B”</b> represents individuals that presented a response to both pathways. (B) Effect of age on the acquisition of antibody responses to the two invasion pathways.</p

Comparison of EBA175 and PfRh2 antibody levels between individuals that switched between invasion inhibition patterns and those with unchanging invasion patterns.

<p>Bars represent the mean antibody levels and SEM. ‘‘Switchers” represents individuals that had fluctuating invasion inhibitory patterns while ‘‘Non-switchers” represents those that had stable invasion inhibitory pattern.</p

Longitudinal invasion inhibitory activity profiles of representative individuals.

<p>Individuals exhibited diverse invasion inhibitory activity against the 3D7 WT relative to the EBA knockout parasites overtime. Individual AD23 inhibited the EBA knockout parasites more than the parental parasite; individual AD50 inhibited the parental parasite more than the EBA knockout parasite lines.</p