Patho-immunological modulatory effect of the human gut microbiome in children with severe P. falciparum infections from Cross River, southern Nigeria

Current malaria control strategies are bedevilled by the development of resistance both by the parasite and vector, hence there is a need to look at other alternative measures that can be applied to mitigate the burden of malaria. This study will employ metagenomics, meta-transcriptomics, and dual RNA-sequencing of the human gut microbiome (HGM) and P. falciparum in understanding the interplay between the HGM and malaria and how this impacts the progression of the disease. This study has the potential to design possible future probiotic interventions for malaria control.

Twenty children, after obtaining parental consent, would be enrolled in this study. Participants would be stratified into two age groups (from 6 months to less than 3years and above 3 years to less than 5 years) and recruited from Akpabuyo General Hospital. Blood and stool samples would be collected on day 0, pre-treatment, from children. Rapidly, all blood samples would be subjected to P. falciparum rapid diagnostic kit (PfHRP-II) and subsequently by microscopy. Those found positive by both diagnostic techniques would be observed, and the same samples (blood and stool) collected on days 1, 2, 3, 7, 1,4 and 21 post-treatments. All collected samples would be shipped to Liverpool School of Tropical Medicine Isolation of DNA will be carried out on all collected blood and stool samples using commercially available kits. Quantitative PCR targeting the highly conserved varATS P. falciparum gene will be used to specifically validate the P. falciparum mono-infections. From the total 140 samples collected from all the participants at all time points, only 70 validated P. falciparum mono-infections, which would be representative of the study population, would be subjected to WGS using the Nextera XT Kit on the Illumina MiSeq platform. Following this, only the corresponding stool of these 70 blood isolates would be processed for sequencing using the variable region (V3-V4) of the 16S rRNA of microbes. While ELISA analysis targeting the a-gal specific IgM and IgG specific antibodies would be carried out on all the one hundred and forty samples, comparative analysis to evaluate the interaction of the human gut microbiome on severe malaria and immune response would only be carried out on all corresponding 70 samples.

EDCTP Grants

Basic Science
Vulnerable Populations

Dec 2020 — Sep 2022

$83,552

The Gambia