title: Successful human infection with P. falciparum using three aseptic Anopheles stephensi mosquitoes: a new model for controlled human malaria infection.
abstract: Controlled human malaria infection (CHMI) is a powerful method for assessing the efficacy of anti-malaria vaccines and drugs targeting pre-erythrocytic and erythrocytic stages of the parasite. CHMI has heretofore required the bites of 5 Plasmodium falciparum (Pf) sporozoite (SPZ)-infected mosquitoes to reliably induce Pf malaria. We reported that CHMI using the bites of 3 PfSPZ-infected mosquitoes reared aseptically in compliance with current good manufacturing practices (cGMP) was successful in 6 participants. Here, we report results from a subsequent CHMI study using 3 PfSPZ-infected mosquitoes reared aseptically to validate the initial clinical trial. We also compare results of safety, tolerability, and transmission dynamics in participants undergoing CHMI using 3 PfSPZ-infected mosquitoes reared aseptically to published studies of CHMI using 5 mosquitoes. Nineteen adults aged 18-40 years were bitten by 3 Anopheles stephensi mosquitoes infected with the chloroquine-sensitive NF54 strain of Pf. All 19 participants developed malaria (100%); 12 of 19 (63%) on Day 11. The mean pre-patent period was 258.3 hours (range 210.5-333.8). The geometric mean parasitemia at first diagnosis by microscopy was 9.5 parasites/µL (range 2-44). Quantitative polymerase chain reaction (qPCR) detected parasites an average of 79.8 hours (range 43.8-116.7) before microscopy. The mosquitoes had a geometric mean of 37,894 PfSPZ/mosquito (range 3,500-152,200). Exposure to the bites of 3 aseptically-raised, PfSPZ-infected mosquitoes is a safe, effective procedure for CHMI in malaria-naïve adults. The aseptic model should be considered as a new standard for CHMI trials in non-endemic areas. Microscopy is the gold standard used for the diagnosis of Pf malaria after CHMI, but qPCR identifies parasites earlier. If qPCR continues to be shown to be highly specific, and can be made to be practical, rapid, and standardized, it should be considered as an alternative for diagnosis.
  
  
  BACKGROUND
  ClinicalTrials.gov NCT00744133 NCT00744133.
authors.affiliation: Howard Hughes Medical Institute/Center for Vaccine Development, University of Maryland School of Medicine, Baltimore, Maryland, United States of America.
authors.forename: Matthew B
authors.initials: MB
authors.lastname: Laurens
authors.forename: Peter
authors.initials: P
authors.lastname: Billingsley
authors.forename: Adam
authors.initials: A
authors.lastname: Richman
authors.forename: Abraham G
authors.initials: AG
authors.lastname: Eappen
authors.forename: Matthew
authors.initials: M
authors.lastname: Adams
authors.forename: Tao
authors.initials: T
authors.lastname: Li
authors.forename: Sumana
authors.initials: S
authors.lastname: Chakravarty
authors.forename: Anusha
authors.initials: A
authors.lastname: Gunasekera
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Jacob
authors.forename: B Kim Lee
authors.initials: BK
authors.lastname: Sim
authors.forename: Robert
authors.initials: R
authors.lastname: Edelman
authors.forename: Christopher V
authors.initials: CV
authors.lastname: Plowe
authors.forename: Stephen L
authors.initials: SL
authors.lastname: Hoffman
authors.forename: Kirsten E
authors.initials: KE
authors.lastname: Lyke
mesh_terms: D000293:Adolescent; D000328:Adult; D000818:Animals; D000852:Anopheles / Q000469:parasitology*; D005260:Female; D006801:Humans; D016778:Malaria, Falciparum / Q000635:transmission*; D008297:Male; D010963:Plasmodium falciparum / Q000472:pathogenicity*; D016133:Polymerase Chain Reaction; D055815:Young Adult