TY - JOUR
T1 - Dissection-independent production of Plasmodium sporozoites from whole mosquitoes
AU - Blight, Joshua
AU - Sala, Katarzyna A.
AU - Atcheson, Erwan
AU - Kramer, Holger
AU - Turabi, Aadil El
AU - Real, Eliana
AU - Dahalan, Farah A.
AU - Bettencourt, Paulo
AU - Dickinson-Craig, Emma
AU - Alves, Eduardo
AU - Salman, Ahmed M.
AU - Janse, Chris J.
AU - Ashcroft, Frances M.
AU - Hill, Adrian Vs
AU - Reyes-Sandoval, Arturo
AU - Blagborough, Andrew M.
AU - Baum, Jake
N1 - Funding Information:
This work is dedicated to the memory of Shahid M Kahn for his contributions to whole sporozoite vaccinology and for his generosity of spirit, collaborating on this work. We thank Alex Fyfe and Mark Tunnicliff for maintaining the mosquito colony, Andrew Worth for help with FACS sorting, Mark Shipman and Andreas Bruckbauer for help with microscopy, and Stephen Rothery for writing the script for automated image analysis. We also thank Annie Yang for advice on P. falciparum in vitro hepatocyte assays. We gratefully acknowledge InfraVec for supporting our P. falciparum research, in particular Roel Heutink and Geert-Jan Van for provision of infected mosquitoes required in the testing phases of this work. Research was directly supported by Wellcome (Investigator Award 100993/Z/13/Z, J Baum and Career Development Fellowship 097395/Z/11/Z, A Reyes-Sandoval), the Bill & Melinda Gates Foundation (OPP1200274, J Baum), and an Medical Research Concil (MRC) research training and support grant (1240480, J Blight). AM Blag-borough thanks the MRC (New Investigator Research grant MR/N00227X/1), PATH-MVI, Isaac Newton Trust, Wellcome Trust Institutional Strategic Support Fund and, University of Cambridge Junior Research Fellowship Scheme for funding. E Alves was funded by CAPES (Program Science without Borders, CSF-2361/13-2). FM Ashcroft and H Kramer thank Wellcome (OXION Strategic award 084655/Z/08/Z) for support. Microscopy work was supported through Mark Shipman in the Ludwig Institute at Oxford and the Facility for Imaging by Light Microscopy at Imperial College London, supported by previous funding from Wellcome (grant 104931/Z/14/Z), and the Biotechnology and Biological Sciences Research Council, UK (grant BB/L015129/1). This project has received resources funded by the European Union’s Horizon 2020 research and innovation program under grant agreement No 731060 (Infravec2).
Publisher Copyright:
© 2021 Rockefeller University Press. All rights reserved.
PY - 2021/7
Y1 - 2021/7
N2 - Progress towards a protective vaccine against malaria remains slow. To date, only limited protection has been routinely achieved following immunisation with either whole-parasite (sporozoite) or subunit-based vaccines. One major roadblock to vaccine progress, and to pre-erythrocytic parasite biology in general, is the continued reliance on manual salivary gland dissection for sporozoite isolation from infected mosquitoes. Here, we report development of a multi-step method, based on batch processing of homogenised whole mosquitoes, slurry, and density-gradient filtration, which combined with free-flow electrophoresis rapidly produces a pure, infective sporozoite inoculum. Human-infective Plasmodium falciparum and rodent-infective Plasmodium berghei sporozoites produced in this way are two-To threefold more infective than salivary gland dissection sporozoites in in vitro hepatocyte infection assays. In an in vivo rodent malaria model, the same P. berghei sporozoites confer sterile protection from mosquito-bite challenge when immunisation is delivered intravenously or 60 70% protection when delivered intramuscularly. By improving purity, infectivity, and immunogenicity, this method represents a key advancement in capacity to produce researchgrade sporozoites, which should impact delivery of a wholeparasite based malaria vaccine at scale in the future.
AB - Progress towards a protective vaccine against malaria remains slow. To date, only limited protection has been routinely achieved following immunisation with either whole-parasite (sporozoite) or subunit-based vaccines. One major roadblock to vaccine progress, and to pre-erythrocytic parasite biology in general, is the continued reliance on manual salivary gland dissection for sporozoite isolation from infected mosquitoes. Here, we report development of a multi-step method, based on batch processing of homogenised whole mosquitoes, slurry, and density-gradient filtration, which combined with free-flow electrophoresis rapidly produces a pure, infective sporozoite inoculum. Human-infective Plasmodium falciparum and rodent-infective Plasmodium berghei sporozoites produced in this way are two-To threefold more infective than salivary gland dissection sporozoites in in vitro hepatocyte infection assays. In an in vivo rodent malaria model, the same P. berghei sporozoites confer sterile protection from mosquito-bite challenge when immunisation is delivered intravenously or 60 70% protection when delivered intramuscularly. By improving purity, infectivity, and immunogenicity, this method represents a key advancement in capacity to produce researchgrade sporozoites, which should impact delivery of a wholeparasite based malaria vaccine at scale in the future.
UR - http://www.scopus.com/inward/record.url?scp=85108362153&partnerID=8YFLogxK
U2 - 10.26508/lsa.202101094
DO - 10.26508/lsa.202101094
M3 - Article
C2 - 34135099
AN - SCOPUS:85108362153
SN - 2575-1077
VL - 4
JO - Life Science Alliance
JF - Life Science Alliance
IS - 7
M1 - e202101094
ER -