Cláudia Fançony

Background: A multicomponent vaccine is increasingly recognized as a necessary tool to control and eliminate malaria globally. However, genetic diversity in Plasmodium falciparum poses a significant challenge, particularly for blood-stage antigens associated with protective immunity. Individuals in endemic regions acquire clinical immunity without exposure to all antigenic variants, suggesting that a deeper understanding of antigen diversity could inform more effective vaccine strategies. Methods: The genetic diversity of the polymorphic gene locus for merozoite surface protein 2 was assessed with nested polymerase chain reaction and capillary electrophoresis fragment sizing in P falciparum isolates from 34 countries across sub-Saharan Africa. High fidelity long-read sequencing was used to investigate sequence diversity within msp2 variants. Results: We successfully genotyped msp2 from 2761 P falciparum–positive samples using capillary electrophoresis, identifying 412 distinct msp2 size variants. Several msp2 size variants were consistently overrepresented across geographical regions, transmission intensities, and time points. These variants comprised multiple unique sequences, of which several were geographically and temporally widespread. Nucleotide and amino acid sequences showed high diversity without geographical clustering. Conclusions: This study provides a comprehensive assessment of msp2 diversity across sub-Saharan Africa. The widespread prevalence of certain variants of this highly polymorphic antigen suggests possible structural constraints or selective advantages for the parasite. Maintenance of specific gene lengths and sequences across diverse settings highlights their potential as targets for next-generation, multicomponent malaria vaccines.

Genetic diversity in Plasmodium falciparum poses a significant challenge to malaria control and elimination. This is particularly important for developing fully efficacious vaccines, which should include valuable blood stage antigens. Several antigen candidates are highly diverse and require further understanding. We surveyed the genetic diversity of the highly polymorphic merozoite surface protein 2 (MSP2) in 2761 P. falciparum isolates collected across Sub-Saharan Africa. Using PCR-based genotyping and long-read sequencing, we identified extensive diversity among msp2 size variants and sequences. Some size variants were more prevalent than others across different geographical regions, transmission intensities, and time points. These variants comprised multiple unique sequences, of which several were geographically and temporally widespread. Our study reveals greater msp2 sequence diversity than previously known, while also identifying interesting similarities in sequence and gene length across Sub-Saharan Africa. These findings support the further exploration of common msp2 variants in relation to parasite virulence and vaccine development.

Background: Decreased efficacy of artemether-lumefantrine, the globally most used antimalarial, has recently emerged in Africa. Methods: An efficacy trial was carried out based on directly observed artemether-lumefantrine therapy at Bengo, Northern Angola. One-hundred Plasmodium falciparum uncomplicated malaria patients (2–10 years old) were enrolled, hospitalized for the treatment period, and followed up for 42 days. Polymerase chain reaction (PCR) correction was performed with pfmsp1/2 plus glurp, with analysis considering 2 or 3 coincident markers. Infections were tested by quantitative PCR (qPCR) for pfmdr1 copy number (pfmdr1×N), a potential P. falciparum marker of lumefantrine resistance previously identified in the region. In vitro clone mixtures were built and used to determine the relation between qPCR copy number scores and actual intrainfection quantitative fractions of pfmdr1×N. Results: We observed a significant posttreatment selection of gene amplification, suggesting a role in the parasite in vivo response to this drug. pfmdr1×2 qPCR scores of 1.3, 1.4, and 1.5 were determined to correspond to 15%, 25%, and 35% intrainfection rates. Patients carrying infections with a score ≥1.4 at baseline were linked to decreased artemether-lumefantrine day 42 efficacy (79% vs 97% single-copy pfmdr1). All infections were pfmdr1 N86 carriers and no pfk13 mutations were found. Conclusions: Our study suggests pfmdr1×N as a marker of P. falciparum in vivo response to lumefantrine in Africa, while indicating patients carrying infections with a pretreatment pfmdr1×N score ≥1.4 before treatment are a group experiencing decreased artemether-lumefantrine performance.

Schistosomiasis and soil-transmitted helminths (STH) infections are major public health problems. We aimed to study the 6-mo impact of mass drug administration with praziquantel and albendazole on urinary schistosomiasis and STH. We examined children (aged 2–15 y) from one hamlet, who provided urine and faeces samples at baseline (n=197), 1 mo (n=102) and 6 mo (n=92); 67 completed the protocol. At baseline, 47/67 (70.1%) children presented Schistosoma haematobium (75.8% in the baseline total sample) and 12/67 (17.9%) with STH (30.5% in the initial sample, p=0.010). Among the children, 47.3% had heavy Schistosoma haematobium infection. The most frequent STH was Trichuris trichiura in 9.0%. We also found Hymenolepis nana (13.2%) and Plasmodium falciparum (9.1%) infections and anaemia (82.1%). One mo after chemotherapy there was a significant (p=0.013) reduction of Schistosoma haematobium prevalence (23.5%) and a high egg reduction rate (86.9%). Considering the sample of 67 children, the mean egg concentration was 498 at baseline, 65 at 1 mo and 252 at 6 mo (p<0.05). We also observed a reduction in STH infections, 50% in Ascaris lumbricoides, 33.3% in T. trichiura and 50% in hookworms. At 6 mo, the prevalence of Schistosoma haematobium (76.1%) was similar to the baseline and the STH reduction was not significant. Longitudinal studies have reported many losses in these settings, but we were able to show that mass drug administration for control of schistosomiasis and STH present low effectiveness, that reinfections occur rapidly and that stand alone anthelmintic therapy is not a sustainable choice.