A multiplex assay for the sensitive detection and quantification of male and female Plasmodium falciparum gametocytes

Meerstein-Kessel, Lisette; Andolina, Chiara; Carrió, Elvira; Mahamar, Almahamoudou; Sawa, Patrick; Diawara, Halimatou; Vegte‐Bolmer, Marga van de; Stone, Will; Collins, Katharine A.; Schneider, P.; Dicko, Alassane; Drakeley, Chris; Felger, Ingrid; Voss, Till S.; Lanke, Kjerstin; Bousema, Teun

doi:10.1186/s12936-018-2584-y

Cited by 55 publications

(62 citation statements)

References 45 publications

(87 reference statements)

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“…Overall, this trend held true for the 158 female-enriched transcripts in cluster 9, including those encoding canonical female markers, e.g. osmiophilic body protein g377 (pf3d7_1250100) (62,63), late-stage antigen pfs25 (pf3d7_1031000) (35,63) and ccp1-3 (pf3d7_1475500, pf3d7_1455800, pf3d7_1407000) (35,63) and ccp4 (pf3d7_0903800) that was recently used to reliably type male and female gametocytes in late-stage gametocytes (64). We also detect a small subset of female-enriched transcripts (pf3d7_0918700, imp2 (pf3d7_0730400), pf3d7_1007800, pf3d7_1466800, pf3d7_1146800, obc13 (pf3d7_1214800)) that are expressed earlier in gametocyte development (Fig 4B) and could potentially be important for female development before morphological differences are apparent.…”

Section: Resultsmentioning

confidence: 96%

Hierarchical transcriptional control regulatesPlasmodium falciparumsexual differentiation

Biljon

Wyk

Painter³

et al. 2019

Preprint

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18Although malaria is characterized by cyclic waves of infection by asexual parasites, 19 only sexual gametocyte forms of the malaria parasite can be transmitted between the 20 infected human and the mosquito host. Here we provide a high-resolution 21 transcriptome of P. falciparum as it commits to and develops through 22 gametocytogenesis and prepares for transmission to the mosquito vector. As 23 anticipated, the gametocyte-associated transcriptome is significantly different from 24 that of the asexual blood cycle, and a dynamic shift in gene expression characterizes 25 early, intermediate and late-stage gametocyte development. Moreover, we show a 26 differential timing in the expression of sex-specific transcripts and we highlight early 27 male and female gametocyte markers for functional evaluation. The striking temporal 28 transcript abundance profiles suggest that strict transcriptional control underlies the 29 10-day gametocyte developmental program in P. falciparum. We identify putative 30 regulators of transcriptional dynamics, including epigenetic mechanisms driving active 31 repression of processes typically associated with proliferation and ApiAP2 32 transcription factors expressed during gametocyte maturation. The gametocyte 33 transcriptome serves as the blueprint for sexual differentiation at the mRNA transcript 34 level and will be a rich resource for future functional studies on this critical stage of 35 Plasmodium development, as the intraerythrocytic transcriptome has been for our 36 understanding of the 48-hour asexual cycle. 3 37 Author Summary 38The ability of the malaria parasite Plasmodium falciparum to smoothly transition 39 between the human and the female mosquito hosts is one of the hallmarks of the 40 pathogenicity of the disease. Only sexually dimorphic, mature gametocyte stages of 41 the parasite life cycle can complete the human-to-mosquito transmission task. 42However, we do not clearly understand how these gametocytes develop and mature. 43Here, we report the highest resolution investigation of the gametocyte's transcriptional 44 profile during development and differentiation. This information sheds light on several 45 possible regulators that mediate parasite maturation into transmission-competent 46 stage V gametocytes. We find that 15% of the parasite's transcripts are 47 developmentally regulated within three distinct phases of sexual development, each 48 corresponding to essential biological events allowing for regulated and staged 49 differentiation. Our study lays the foundation for future projects aiming to 50 therapeutically target these transmissible stages and comprises an invaluable 51 resource for the study of malaria parasite pathogenesis. 52 53 54 55 4 57 Introduction 58 Sustained malaria prevalence is ensured through continued human-to-mosquito 59 transmission of Plasmodium parasites with Plasmodium falciparum being the 60 causative agent of the most severe form of the disease in humans (1). The complex 61 life cycle of P. falciparum encompasses development in ...

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Section: Resultsmentioning

confidence: 96%

Hierarchical transcriptional control regulatesPlasmodium falciparumsexual differentiation

Biljon

Wyk

Painter³

et al. 2019

Preprint

View full text Add to dashboard Cite

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“…Overall, this trend held true for the 158 female-enriched transcripts in cluster 9, including those encoding canonical female markers, e.g. osmiophilic body protein g377 (pf3d7_1250100) [65,66], late-stage antigen pfs25 (pf3d7_1031000) [35,66] and ccp1-3 (pf3d7_ 1475500, pf3d7_1455800, pf3d7_1407000) [35,66] and ccp4 (pf3d7_0903800) that was recently used to reliably type male and female gametocytes in late-stage gametocytes [67]. We also detect a small subset of female-enriched transcripts (pf3d7_0918700, imp2 (pf3d7_0730400), pf3d7_1007800, pf3d7_1466800, pf3d7_1146800, obc13 (pf3d7_1214800)) that are expressed earlier in gametocyte development (Fig.…”

Section: Transcriptional Patterns Characterize Distinct Transitions Imentioning

confidence: 86%

Hierarchical transcriptional control regulates Plasmodium falciparum sexual differentiation

Biljon

Wyk

Painter

et al. 2019

Preprint

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Background: Malaria pathogenesis relies on sexual gametocyte forms of the malaria parasite to be transmitted between the infected human and the mosquito host but the molecular mechanisms controlling gametocytogenesis remains poorly understood. Here we provide a high-resolution transcriptome of Plasmodium falciparum as it commits to and develops through gametocytogenesis. Results : The gametocyte-associated transcriptome is significantly different from that of the asexual parasites, with dynamic gene expression shifts characterizing early, intermediate and late-stage gametocyte development and results in differential timing for sex-specific transcripts. The transcriptional dynamics suggest strict transcriptional control during gametocytogenesis in P. falciparum, which we propose is mediated by putative regulators including epigenetic mechanisms (driving active repression of proliferation-associated processes) and a cascade-like expression of ApiAP2 transcription factors. Conclusions : The gametocyte transcriptome serves as the blueprint for sexual differentiation and will be a rich resource for future functional studies on this critical stage of Plasmodium development, as the intraerythrocytic transcriptome has been for our understanding of the asexual cycle.

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“…Mosquito midguts were examined for the presence of parasite developmental stages, oocysts, by two independent microscopists. estimates of male and female gametocytes were adjusted for background signal from asexual parasites 56 . qPCR targeting pfap2-g 57 and sbp1 58 was performed on day of first treatment and the ratio of pfap2-g:sbp1 used as an indication of the proportion of sexually committed ring-stage parasites 59 .…”

Section: Mosquito Feeding Assaysmentioning

confidence: 99%

Increased gametocyte production and mosquito infectivity in chronic versus incident Plasmodium falciparum infections

Barry

Bradley

Guelbeogo

et al. 2020

Preprint

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30We longitudinally assessed P. falciparum parasite kinetics, gametocyte production and infectivity 31 in incident infections that were naturally acquired following infection clearance and in chronic 32 asymptomatic infections in Burkina Faso. 92% (44/48) of the incident cohort developed symptoms 33 and were treated within 35 days, compared to 23% (14/60) of the chronic cohort. All but two 34 individuals with chronic infection were gametocytaemic at enrollment, whereas only 35% (17/48) 35in the incident cohort developed gametocytes within 35 days. The relative abundance of ap2-g 36 transcripts was positively associated with conversion to gametocyte production (i.e. the ratio of 37 gametocytes at day 14 to ring stage parasites at baseline) and was higher in chronic infections. 38Parasite multiplication rate, assessed by daily molecular parasite quantification, was positively 39 associated with prospective gametocyte production. Most incident infections were cleared before 40 gametocyte density was sufficiently high to infect mosquitoes. In contrast, chronic, asymptomatic 41 infections represented a significant source of mosquito infections. If present, gametocytes were 42 significantly less infectious if concurrent with malaria symptoms. Our observations support the 43 notion that malaria transmission reduction may be expediated by enhanced case management, 44 involving both symptom-screening and infection detection.45 46 47 48The epidemiology of P. falciparum transmission stages, gametocytes, is poorly understood. 50Molecular diagnostics show that low density gametocyte carriage is highly prevalent in endemic 51 populations 1 and that many low density infections are infectious to mosquitoes 2 , explaining 52 observations from the 1950s that gametocyte free individuals (as determined by microscopy) 53 frequently infected mosquitoes 3 . Few assessments of the population infectious reservoir of 54 malaria parasites have been undertaken 4,5 , particularly with tools capable of detecting low 55 parasite and gametocyte densities 6,7 . Fewer still have been able to assess the association of 56 infectivity with factors other than parasite density, despite clear evidence that gametocyte 57 maturity 8,9 , intrinsic parasite factors 10,11 , human genetic factors 12 , and human clinical and immune 58 responses 13-15 can have significant influence on the effectiveness of parasite transmission. 59The association between malaria symptoms and gametocyte dynamics is particularly poorly 60 understood. Among similarly immune individuals, higher densities of pathogenic, asexual stage 61 malaria parasites are commonly associated with the presentation of symptoms 16 . Because 62 gametocytes develop from asexual parasites, individuals with clinical malaria and higher parasite 63 densities may have more gametocytes and be more likely to infect mosquitoes 17,18 . On the other 64 hand, the slower development of gametocytes may result in higher densities later in infections. 65This would result in higher gametocyte densities in infecti...

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A multiplex assay for the sensitive detection and quantification of male and female Plasmodium falciparum gametocytes

Cited by 55 publications

References 45 publications

Hierarchical transcriptional control regulatesPlasmodium falciparumsexual differentiation

Hierarchical transcriptional control regulatesPlasmodium falciparumsexual differentiation

Hierarchical transcriptional control regulates Plasmodium falciparum sexual differentiation

Increased gametocyte production and mosquito infectivity in chronic versus incident Plasmodium falciparum infections

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