Doumbo, S. N. | Search bioRxiv

bioRxiv2025

Seasonal transcriptional profiling of malaria parasites using a new single-cell atlas of a Malian Plasmodium falciparum isolate

Asymptomatic persistence is crucial for the survival of malaria parasites in seasonal transmission settings, where transmission halts during dry months, while mosquitos are scarce and new infections rare. How Plasmodium falciparum avoids causing host symptoms or being cleared is not fully understood. Parasites that persist through the dry season circulate longer within the [~]48h replication cycle compared to parasites isolated from clinical cases, where only early stages circulate. The mechanisms promoting increased circulation and avirulent infections bridging two wet seasons remain unclear. We generated a single-cell RNA-seq reference atlas with P. falciparum erythrocytic stages of a recently adapted isolate, and with it defined the developmental stage of over 25000 parasites from six asymptomatic children at the dry-wet season transition, and nine children with clinical malaria in the wet season. Our data confirms that older parasites circulate in the dry season, and reveals transcriptional modulation of exported proteins that affect cytoadhesion of infected erythrocytes, which we validated by immuno-fluorescence microscopy.

molecular biology↗

bioRxiv2024

Plasmodium falciparum expresses fewer var genes at lower levels during asymptomatic dry season infections than clinical malaria cases

In seasonal transmission areas, clinical malaria occurs during the wet season when mosquitoes are present, while in the dry season, malaria transmission is interrupted and clinical cases are rare. In Mali, Plasmodium falciparum can persist in low parasitaemic asymptomatic individuals through the six-month dry season and shows reduced cytoadhesion of infected erythrocytes, evidenced by the circulation of further developed parasite stages compared to clinical malaria cases. How prolonged circulation of infected erythrocytes is achieved remains unknown. Here, we explored var gene expression in subclinical infections and clinical malaria cases of Malian children, collected during the dry and wet seasons. We sequenced expressed var DBL-tags, used bioinformatic tools to predict their domain composition, binding phenotype and upstream sequence type; and determined their relationship to seasonality and clinical presentation. We found that parasites of asymptomatic infections expressed fewer var genes, with a larger proportion of var transcripts attributed to one or a few vars. In contrast, clinical cases exhibited expression of many var genes at lower proportions. We found that parasites of asymptomatic carriers expressed a mixture of CD36- and EPCR-binding PfEMP1, which changed over time. We confirmed that vars encoding CD36-binding PfEMP1 dominated in non-severe malaria cases, and found no significant difference in expressed var types between dry and wet seasons. Asymptomatic carriers were older, had higher titers of anti-P. falciparum antibodies, and broader reactivity to PfEMP1, suggesting that host immunity was the main determinant limiting var transcript variation in asymptomatic carriers. However, by RNAseq and qRT-PCR we also observed significantly higher total var transcript levels in malaria cases compared to asymptomatic carriers, suggesting that in addition to the parasites switching and the hosts immune selection of expressed var genes, parasites able to sustain long-term infections may be poised for reduced PfEMP1 expression.

microbiology↗