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01-12-2021 | Malaria | Research article

Safety, infectivity and immunogenicity of a genetically attenuated blood-stage malaria vaccine

Authors: Rebecca Webster, Silvana Sekuloski, Anand Odedra, Stephen Woolley, Helen Jennings, Fiona Amante, Katharine R. Trenholme, Julie Healer, Alan F. Cowman, Emily M. Eriksson, Priyanka Sathe, Jocelyn Penington, Adam J. Blanch, Matthew W. A. Dixon, Leann Tilley, Michael F. Duffy, Alister Craig, Janet Storm, Jo-Anne Chan, Krystal Evans, Anthony T. Papenfuss, Louis Schofield, Paul Griffin, Bridget E. Barber, Dean Andrew, Michelle J. Boyle, Fabian de Labastida Rivera, Christian Engwerda, James S. McCarthy

Published in: BMC Medicine | Issue 1/2021

Abstract

Background

There is a clear need for novel approaches to malaria vaccine development. We aimed to develop a genetically attenuated blood-stage vaccine and test its safety, infectivity, and immunogenicity in healthy volunteers. Our approach was to target the gene encoding the knob-associated histidine-rich protein (KAHRP), which is responsible for the assembly of knob structures at the infected erythrocyte surface. Knobs are required for correct display of the polymorphic adhesion ligand P. falciparum erythrocyte membrane protein 1 (PfEMP1), a key virulence determinant encoded by a repertoire of var genes.

Methods

The gene encoding KAHRP was deleted from P. falciparum 3D7 and a master cell bank was produced in accordance with Good Manufacturing Practice. Eight malaria naïve males were intravenously inoculated (day 0) with 1800 (2 subjects), 1.8 × 10⁵ (2 subjects), or 3 × 10⁶ viable parasites (4 subjects). Parasitemia was measured using qPCR; immunogenicity was determined using standard assays. Parasites were rescued into culture for in vitro analyses (genome sequencing, cytoadhesion assays, scanning electron microscopy, var gene expression).

Results

None of the subjects who were administered with 1800 or 1.8 × 10⁵ parasites developed parasitemia; 3/4 subjects administered 3× 10⁶ parasites developed significant parasitemia, first detected on days 13, 18, and 22. One of these three subjects developed symptoms of malaria simultaneously with influenza B (day 17; 14,022 parasites/mL); one subject developed mild symptoms on day 28 (19,956 parasites/mL); and one subject remained asymptomatic up to day 35 (5046 parasites/mL). Parasitemia rapidly cleared with artemether/lumefantrine. Parasitemia induced a parasite-specific antibody and cell-mediated immune response. Parasites cultured ex vivo exhibited genotypic and phenotypic properties similar to inoculated parasites, although the var gene expression profile changed during growth in vivo.

Conclusions

This study represents the first clinical investigation of a genetically attenuated blood-stage human malaria vaccine. A P. falciparum 3D7 kahrp– strain was tested in vivo and found to be immunogenic but can lead to patent parasitemia at high doses.

Trial registration

Australian New Zealand Clinical Trials Registry (number: ACTRN12617000824369; date: 06 June 2017).

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Duffy PE, Patrick GJ. Malaria vaccines since 2000: progress, priorities, products. NPJ Vaccines. 2020;5(1):48. https://doi.org/10.1038/s41541-020-0196-3.CrossRefPubMedPubMedCentral

Clinical Trials Partnership RTSS. Efficacy and safety of RTS,S/AS01 malaria vaccine with or without a booster dose in infants and children in Africa: final results of a phase 3, individually randomised, controlled trial. Lancet. 2015;386(9988):31–45. https://doi.org/10.1016/S0140-6736(15)60721-8.CrossRef

Malaria vaccine implementation programme (MVIP): World Health Organization; [updated March 2, 2020. Available from: https://www.who.int/news-room/q-a-detail/malaria-vaccine-implementation-programme.

Stanisic DI, Good MF. Whole organism blood stage vaccines against malaria. Vaccine. 2015;33(52):7469–75. https://doi.org/10.1016/j.vaccine.2015.09.057.CrossRefPubMed

Epstein JE, Tewari K, Lyke KE, Sim BK, Billingsley PF, Laurens MB, et al. Live attenuated malaria vaccine designed to protect through hepatic CD8(+) T cell immunity. Science. 2011;334(6055):475–80. https://doi.org/10.1126/science.1211548.CrossRefPubMed

Kublin JG, Mikolajczak SA, Sack BK, Fishbaugher ME, Seilie A, Shelton L, et al. Complete attenuation of genetically engineered Plasmodium falciparum sporozoites in human subjects. Sci Transl Med. 2017;9:eaad9099.CrossRef

Reuling IJ, Mendes AM, de Jong GM, Fabra-Garcia A, Nunes-Cabaco H, van Gemert GJ, et al. An open-label phase 1/2a trial of a genetically modified rodent malaria parasite for immunization against Plasmodium falciparum malaria. Sci Transl Med. 2020;12:eaay2578.CrossRef

Roestenberg M, Walk J, van der Boor SC, Langenberg MCC, Hoogerwerf MA, Janse JJ, et al. A double-blind, placebo-controlled phase 1/2a trial of the genetically attenuated malaria vaccine PfSPZ-GA1. Sci Transl Med. 2020;12:eaaz5629.CrossRef

Draper SJ, Sack BK, King CR, Nielsen CM, Rayner JC, Higgins MK, et al. Malaria vaccines: recent advances and new horizons. Cell Host Microbe. 2018;24(1):43–56. https://doi.org/10.1016/j.chom.2018.06.008.CrossRefPubMedPubMedCentral

10.

Stanisic DI, Fink J, Mayer J, Coghill S, Gore L, Liu XQ, et al. Vaccination with chemically attenuated Plasmodium falciparum asexual blood-stage parasites induces parasite-specific cellular immune responses in malaria-naive volunteers: a pilot study. BMC Med. 2018;16(1):184. https://doi.org/10.1186/s12916-018-1173-9.CrossRefPubMedPubMedCentral

11.

Baruch DI, Rogerson SJ, Cooke BM. Asexual blood stages of malaria antigens: cytoadherence. Chem Immunol. 2002;80:144–62. https://doi.org/10.1159/000058839.CrossRefPubMed

12.

Howard RJ, Gilladoga AD. Molecular studies related to the pathogenesis of cerebral malaria. Blood. 1989;74(8):2603–18. https://doi.org/10.1182/blood.V74.8.2603.2603.CrossRefPubMed

13.

Rowe JA, Moulds JM, Newbold CI, Miller LH. P. falciparum rosetting mediated by a parasite-variant erythrocyte membrane protein and complement-receptor 1. Nature. 1997;388:292–5.CrossRef

14.

Gonzales SJ, Reyes RA, Braddom AE, Batugedara G, Bol S, Bunnik EM. Naturally acquired humoral immunity against Plasmodium falciparum malaria. Front Immunol. 2020;11:594653. https://doi.org/10.3389/fimmu.2020.594653.CrossRefPubMedPubMedCentral

15.

D'Ombrain MC, Voss TS, Maier AG, Pearce JA, Hansen DS, Cowman AF, et al. Plasmodium falciparum erythrocyte membrane protein-1 specifically suppresses early production of host interferon-gamma. Cell Host Microbe. 2007;2(2):130–8. https://doi.org/10.1016/j.chom.2007.06.012.CrossRefPubMed

16.

Su XZ, Heatwole VM, Wertheimer SP, Guinet F, Herrfeldt JA, Peterson DS, et al. The large diverse gene family var encodes proteins involved in cytoadherence and antigenic variation of Plasmodium falciparum-infected erythrocytes. Cell. 1995;82(1):89–100. https://doi.org/10.1016/0092-8674(95)90055-1.CrossRefPubMed

17.

Langreth SG, Peterson E. Pathogenicity, stability, and immunogenicity of a knobless clone of Plasmodium falciparum in Colombian owl monkeys. Infect Immun. 1985;47(3):760–6. https://doi.org/10.1128/iai.47.3.760-766.1985.CrossRefPubMedPubMedCentral

18.

Crabb BS, Cooke BM, Reeder JC, Waller RF, Caruana SR, Davern KM, et al. Targeted gene disruption shows that knobs enable malaria-infected red cells to cytoadhere under physiological shear stress. Cell. 1997;89(2):287–96. https://doi.org/10.1016/S0092-8674(00)80207-X.CrossRefPubMed

19.

Pawliw R, Farrow R, Sekuloski S, Jennings H, Healer J, Phuong T, et al. A bioreactor system for the manufacture of a genetically modified Plasmodium falciparum blood stage malaria cell bank for use in a clinical trial. Malar J. 2018;17(1):283. https://doi.org/10.1186/s12936-018-2435-x.CrossRefPubMedPubMedCentral

20.

Rockett RJ, Tozer SJ, Peatey C, Bialasiewicz S, Whiley DM, Nissen MD, et al. A real-time, quantitative PCR method using hydrolysis probes for the monitoring of Plasmodium falciparum load in experimentally infected human volunteers. Malar J. 2011;10(1):48. https://doi.org/10.1186/1475-2875-10-48.CrossRefPubMedPubMedCentral

21.

McCarthy JS, Lotharius J, Ruckle T, Chalon S, Phillips MA, Elliott S, et al. Safety, tolerability, pharmacokinetics, and activity of the novel long-acting antimalarial DSM265: a two-part first-in-human phase 1a/1b randomised study. Lancet Infect Dis. 2017;17(6):626–35. https://doi.org/10.1016/S1473-3099(17)30171-8.CrossRefPubMedPubMedCentral

22.

McCarthy JS, Ruckle T, Elliott SL, Ballard E, Collins KA, Marquart L, et al. A single-dose combination study with the experimental antimalarials artefenomel and DSM265 to determine safety and antimalarial activity against blood-stage Plasmodium falciparum in healthy volunteers. Antimicrob Agents Chemother. 2019;64(1):e01371–19. https://doi.org/10.1128/AAC.01371-19.CrossRefPubMedPubMedCentral

23.

McCarthy JS, Sekuloski S, Griffin PM, Elliott S, Douglas N, Peatey C, et al. A pilot randomised trial of induced blood-stage Plasmodium falciparum infections in healthy volunteers for testing efficacy of new antimalarial drugs. PloS One. 2011;6(8):e21914. https://doi.org/10.1371/journal.pone.0021914.CrossRefPubMedPubMedCentral

24.

Warrell DA, Gilles. HM, editors. Essential Malariology. Fourth ed: CRC Press; 2017.

25.

Collins KA, Ruckle T, Elliott S, Marquart L, Ballard E, Chalon S, et al. DSM265 at 400 milligrams clears asexual stage parasites but not mature gametocytes from the blood of healthy subjects experimentally infected with Plasmodium falciparum. Antimicrob Agents Chemother. 2019;63(4):e01837–18. https://doi.org/10.1128/AAC.01837-18.CrossRefPubMedPubMedCentral

26.

Common Terminology Criteria for Adverse Events (CTCAE) Version 4.03. Bethesda: US Department of Health and Human Services; 2010.

27.

Boyle MJ, Chan JA, Handayuni I, Reiling L, Feng G, Hilton A, et al. IgM in human immunity to Plasmodium falciparum malaria. Sci Adv. 2019;5:eaax4489.CrossRef

28.

Burel JG, Apte SH, Groves PL, Boyle MJ, Langer C, Beeson JG, et al. Dichotomous miR expression and immune responses following primary blood-stage malaria. JCI Insight. 2017;2(15):e93434. https://doi.org/10.1172/jci.insight.93434.CrossRefPubMedCentral

29.

Montes de Oca M, Kumar R, Rivera FL, Amante FH, Sheel M, Faleiro RJ, et al. Type I interferons regulate immune responses in humans with blood-stage Plasmodium falciparum infection. Cell Rep. 2016;17:399–412.CrossRef

30.

Ng SS, Souza-Fonseca-Guimaraes F, Rivera FL, Amante FH, Kumar R, Gao Y, et al. Rapid loss of group 1 innate lymphoid cells during blood stage Plasmodium infection. Clin Transl Immunology. 2018;7(1):e1003. https://doi.org/10.1002/cti2.1003.CrossRefPubMedPubMedCentral

31.

Aurrecoechea C, Brestelli J, Brunk BP, Dommer J, Fischer S, Gajria B, et al. PlasmoDB: a functional genomic database for malaria parasites. Nucleic Acids Res. 2009;37(Database):D539–43. https://doi.org/10.1093/nar/gkn814.CrossRefPubMed

32.

Cameron DL, Schroder J, Penington JS, Do H, Molania R, Dobrovic A, et al. GRIDSS: sensitive and specific genomic rearrangement detection using positional de Bruijn graph assembly. Genome Res. 2017;27(12):2050–60. https://doi.org/10.1101/gr.222109.117.CrossRefPubMedPubMedCentral

33.

Koboldt DC, Zhang Q, Larson DE, Shen D, McLellan MD, Lin L, et al. VarScan 2: somatic mutation and copy number alteration discovery in cancer by exome sequencing. Genome Res. 2012;22(3):568–76. https://doi.org/10.1101/gr.129684.111.CrossRefPubMedPubMedCentral

34.

Langmead B, Salzberg SL. Fast gapped-read alignment with Bowtie 2. Nat Methods. 2012;9(4):357–9. https://doi.org/10.1038/nmeth.1923.CrossRefPubMedPubMedCentral

35.

Scheinin I, Sie D, Bengtsson H, van de Wiel MA, Olshen AB, van Thuijl HF, et al. DNA copy number analysis of fresh and formalin-fixed specimens by shallow whole-genome sequencing with identification and exclusion of problematic regions in the genome assembly. Genome Res. 2014;24(12):2022–32. https://doi.org/10.1101/gr.175141.114.CrossRefPubMedPubMedCentral

36.

Wei Z, Wang W, Hu P, Lyon GJ, Hakonarson H. SNVer: a statistical tool for variant calling in analysis of pooled or individual next-generation sequencing data. Nucleic Acids Res. 2011;39(19):e132. https://doi.org/10.1093/nar/gkr599.CrossRefPubMedPubMedCentral

37.

Madkhali AM, Alkurbi MO, Szestak T, Bengtsson A, Patil PR, Wu Y, et al. An analysis of the binding characteristics of a panel of recently selected ICAM-1 binding Plasmodium falciparum patient isolates. PloS One. 2014;9(10):e111518. https://doi.org/10.1371/journal.pone.0111518.CrossRefPubMedPubMedCentral

38.

Storm J, Jespersen JS, Seydel KB, Szestak T, Mbewe M, Chisala NV, et al. Cerebral malaria is associated with differential cytoadherence to brain endothelial cells. EMBO Mol Med. 2019;11(2):e9164. https://doi.org/10.15252/emmm.201809164.CrossRefPubMedPubMedCentral

39.

McHugh E, Carmo OMS, Blanch A, Looker O, Liu B, Tiash S, et al. Role of Plasmodium falciparum protein GEXP07 in Maurer's cleft morphology, knob architecture, and P. falciparum EMP1 trafficking. mBio. 2020;11:e03320–19.CrossRef

40.

Bachmann A, Petter M, Krumkamp R, Esen M, Held J, Scholz JA, et al. Mosquito passage dramatically changes var gene expression in controlled human Plasmodium falciparum infections. PLoS Pathog. 2016;12(4):e1005538. https://doi.org/10.1371/journal.ppat.1005538.CrossRefPubMedPubMedCentral

41.

Duffy MF, Byrne TJ, Carret C, Ivens A, Brown GV. Ectopic recombination of a malaria var gene during mitosis associated with an altered var switch rate. J Mol Biol. 2009;389(3):453–69. https://doi.org/10.1016/j.jmb.2009.04.032.CrossRefPubMedPubMedCentral

42.

Duffy MF, Caragounis A, Noviyanti R, Kyriacou HM, Choong EK, Boysen K, et al. Transcribed var genes associated with placental malaria in Malawian women. Infect Immun. 2006;74(8):4875–83. https://doi.org/10.1128/IAI.01978-05.CrossRefPubMedPubMedCentral

43.

Tonkin-Hill GQ, Trianty L, Noviyanti R, Nguyen HHT, Sebayang BF, Lampah DA, et al. The Plasmodium falciparum transcriptome in severe malaria reveals altered expression of genes involved in important processes including surface antigen-encoding var genes. PLoS Biol. 2018;16(3):e2004328. https://doi.org/10.1371/journal.pbio.2004328.CrossRefPubMedPubMedCentral

44.

Robinson BA, Welch TL, Smith JD. Widespread functional specialization of Plasmodium falciparum erythrocyte membrane protein 1 family members to bind CD36 analysed across a parasite genome. Mol Microbiol. 2003;47(5):1265–78. https://doi.org/10.1046/j.1365-2958.2003.03378.x.CrossRefPubMed

45.

Woolley S, Fernandez M, Rebelo M, Llewellyn S, Marquart L, Mc Carthy JS. Development and evaluation of a new Plasmodium falciparum 3D7 blood stage malaria cell bank for use in malaria volunteer infection studies. Malar J. 2021;20(1):93. https://doi.org/10.1186/s12936-021-03627-z.CrossRefPubMedPubMedCentral

46.

McCarthy JS. Baker M, O'Rourke P, Marquart L, Griffin P, Hooft van Huijsduijnen R, et al. Efficacy of OZ439 (artefenomel) against early Plasmodium falciparum blood-stage malaria infection in healthy volunteers. J Antimicrob Chemother. 2016;71(9):2620–7. https://doi.org/10.1093/jac/dkw174.CrossRefPubMedPubMedCentral

47.

McCarthy JS, Donini C, Chalon S, Woodford J, Marquart L, Collins KA, et al. A phase 1, placebo controlled, randomised, single ascending dose study and a volunteer infection study to characterize the safety, pharmacokinetics and antimalarial activity of the Plasmodium phosphatidylinositol 4-kinase inhibitor MMV390048. Clin Infect Dis. 2020;ciaa368.

48.

Wockner LF, Hoffmann I, Webb L, Mordmuller B, Murphy SC, Kublin JG, et al. Growth Rate of Plasmodium falciparum: Analysis of Parasite Growth Data From Malaria Volunteer Infection Studies. J Infect Dis. 2020;221(6):963–72. https://doi.org/10.1093/infdis/jiz557.CrossRefPubMed

49.

Frank M, Dzikowski R, Amulic B, Deitsch K. Variable switching rates of malaria virulence genes are associated with chromosomal position. Mol Microbiol. 2007;64(6):1486–98. https://doi.org/10.1111/j.1365-2958.2007.05736.x.CrossRefPubMedPubMedCentral

50.

Arakawa C, Gunnarsson C, Howard C, Bernabeu M, Phong K, Yang E, et al. Biophysical and biomolecular interactions of malaria-infected erythrocytes in engineered human capillaries. Sci Adv. 2020;6:eaay7243.CrossRef

51.

McCarthy JS, Good MF. Whole parasite blood stage malaria vaccines: a convergence of evidence. Hum Vaccin. 2010;6(1):114–23. https://doi.org/10.4161/hv.6.1.10394.CrossRefPubMed

52.

al Yaman F, Genton B, Anders R, Taraika J, Ginny M, Mellor S, et al. Assessment of the role of the humoral response to Plasmodium falciparum MSP2 compared to RESA and SPf66 in protecting Papua New Guinean children from clinical malaria. Parasite Immunol. 1995;17:493–501.CrossRef

53.

Kumar R, Loughland JR, Ng SS, Boyle MJ, Engwerda CR. The regulation of CD4(+) T cells during malaria. Immunol Rev. 2020;293(1):70–87. https://doi.org/10.1111/imr.12804.CrossRefPubMed

54.

Watts RE, Odedra A, Marquart L, Webb L, Abd-Rahman AN, Cascales L, et al. Safety and parasite clearance of artemisinin-resistant Plasmodium falciparum infection: a pilot and a randomised volunteer infection study in Australia. PLoS Med. 2020;17(8):e1003203. https://doi.org/10.1371/journal.pmed.1003203.CrossRefPubMedPubMedCentral

55.

Collins KA, Wang CY, Adams M, Mitchell H, Robinson GJ, Rampton M, et al. A Plasmodium vivax experimental human infection model for evaluating efficacy of interventions. J Clin Invest. 2020;130(6):2920–7. https://doi.org/10.1172/JCI134923.CrossRefPubMedPubMedCentral

56.

Woodford J, Collins KA, Odedra A, Wang C, Jang IK, Domingo GJ, et al. An experimental human blood-stage model for studying Plasmodium malariae infection. J Infect Dis. 2020;221(6):948–55. https://doi.org/10.1093/infdis/jiz102.CrossRefPubMed

Title: Safety, infectivity and immunogenicity of a genetically attenuated blood-stage malaria vaccine
Authors: Rebecca Webster
Silvana Sekuloski
Anand Odedra
Stephen Woolley
Helen Jennings
Fiona Amante
Katharine R. Trenholme
Julie Healer
Alan F. Cowman
Emily M. Eriksson
Priyanka Sathe
Jocelyn Penington
Adam J. Blanch
Matthew W. A. Dixon
Leann Tilley
Michael F. Duffy
Alister Craig
Janet Storm
Jo-Anne Chan
Krystal Evans
Anthony T. Papenfuss
Louis Schofield
Paul Griffin
Bridget E. Barber
Dean Andrew
Michelle J. Boyle
Fabian de Labastida Rivera
Christian Engwerda
James S. McCarthy
Publication date: 01-12-2021
Publisher: BioMed Central
Keyword: Malaria
Published in: BMC Medicine / Issue 1/2021
Electronic ISSN: 1741-7015
DOI: https://doi.org/10.1186/s12916-021-02150-x

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Safety, infectivity and immunogenicity of a genetically attenuated blood-stage malaria vaccine

Abstract

Background

Methods

Results

Conclusions

Trial registration

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Trial registration

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