Nuclear imaging is an important preclinical research tool to study infectious diseases in vivo and could be extended to investigate complex aspects of malaria infections. As such, we report for the first time successful radiolabeling of a novel antibody specific to Plasmodium-infected erythrocytes (IIIB6), its in vitro assessment and molecular imaging in nude mice.
In vitro confocal microscopy was used to determine the stage-specificity of Plasmodium-infected erythrocytes recognised by IIIB6. To enable micro-positron emission tomography (PET)/X-ray computed tomography (CT) imaging, IIIB6 was conjugated to Bz-DFO-NCS and subsequently radiolabeled with zirconium-89. Healthy nude mice were injected with [89Zr]IIIB6, and pharmacokinetics and organ uptake were monitored over 24 h. This was followed by post-mortem animal dissection to determine the biodistribution of [89Zr]IIIB6.
IIIB6 recognised all the relevant stages of Plasmodium falciparum-infected erythrocytes (trophozoites, schizonts and gametocytes) that are responsible for severe malaria pathology. [89Zr]IIIB6-radiolabeling yields were efficient at 84–89 %. Blood pool imaging analysis indicated a pharmacological half-life of 9.6 ± 2.5 h for [89Zr]IIIB6. The highest standard uptake values were determined at 2–6 h in the liver followed by the spleen, kidneys, heart, stomach and lung, respectively. Minimal activity was present in muscle and bone tissues.
In vitro characterization of IIIB6 and pharmacokinetic characterization of [89Zr]IIIB6 revealed that this antibody has potential for future use in Plasmodium-infected mouse models to study malaria in a preclinical in vivo setting with PET/CT imaging.
This is a preview of subscription content, access via your institution.
Buy single article
Instant access to the full article PDF.
Tax calculation will be finalised during checkout.
Subscribe to journal
Immediate online access to all issues from 2019. Subscription will auto renew annually.
Tax calculation will be finalised during checkout.
Cowman AF, Healer J, Marapana D, Marsh K (2016) Malaria: biology and disease. Cell 167:610–624
Mohandas N, An X (2012) Malaria and human red blood cells. Med Microbiol Immunol 201:593–598
Coppens I, Sullivan DJ, Prigge ST (2010) An update on the rapid advances in malaria parasite cell biology. Trends Parasitol 26:305–310
Zinn KR, Chaudhuri TR, Szafran AA, O'Quinn D, Weaver C, Dugger K, Lamar D, Kesterson RA, Wang X, Frank SJ (2008) Noninvasive bioluminescence imaging in small animals. ILAR J 49:103–115
Maude RJ, Barkhof F, Hassan MU et al (2014) Magnetic resonance imaging of the brain in adults with severe falciparum malaria. Malar J 13:1
Vaidyanathan S, Patel CN, Scarsbrook AF, Chowdhury FU (2015) FDG PET/CT in infection and inflammation—current and emerging clinical applications. Clin Radiol 70:787–800
Sathekge M, Maes A, Van de Wiele C (2015) FDG-PET imaging in HIV infection and tuberculosis. Semin Nucl Med 43:349–366
Sathekge M, Goethals I, Maes A, Van De Wiele C (2009) Positron emission tomography in patients suffering from HIV-1 infection. Eur J Nucl Med Mol Imaging 36:1176–1184
Kawai S, Ikeda E, Sugiyama M et al (2006) Enhancement of splenic glucose metabolism during acute malarial infection: correlation of findings of FDG-PET imaging with pathological changes in a primate model of severe human malaria. Am J Trop Med Hyg 74:353–360
Sugiyama M, Ikeda E, Kawai S et al (2004) Cerebral metabolic reduction in severe malaria: fluorodeoxyglucose-positron emission tomography imaging in a primate model of severe human malaria with cerebral involvement. Am J Trop Med Hyg 71:542–545
Zhang XY, Yang ZL, Lu GM, Yang GF, Zhang LJ (2017) PET/MR imaging: new frontier in Alzheimer’s disease and other dementias. Front Mol Neurosci 10:343
Voss SD, Smith SV, DiBartolo N, McIntosh LJ, Cyr EM, Bonab AA, Dearling JLJ, Carter EA, Fischman AJ, Treves ST, Gillies SD, Sargeson AM, Huston JS, Packard AB (2007) Positron emission tomography (PET) imaging of neuroblastoma and melanoma with 64Cu-SarAr immunoconjugates. Proc Natl Acad Sci 104:17489–17493
Martínez LMA, Castillo AX, Falcón VNC et al (2014) Development of 90Y-DOTA-nimotuzumab Fab fragment for radioimmunotherapy. Radioanal Nucl Chem 302:49–56
Christophides GK, Vlachou D, Kafatos FC (2004) Comparative and functional genomics of the innate immune system in the malaria vector Anopheles gambiae. Immunol Rev 198:127–148
Reader J, Botha M, Theron A, Lauterbach SB, Rossouw C, Engelbrecht D, Wepener M, Smit A, Leroy D, Mancama D, Coetzer TL, Birkholtz LM (2015) Nowhere to hide: interrogating different metabolic parameters of Plasmodium falciparum gametocytes in a transmission blocking drug discovery pipeline towards malaria elimination. Malar J 14:213
Lambros C, Vanderberg JP (1979) Synchronization of Plasmodium falciparum erythrocytic stages in culture. J Parasitol 65:418–420
Hoppe H, Verschoor J, Louw A (1991) Plasmodium falciparum: a comparison of synchronisation methods for in vitro cultures. Exp Parasitol 72:464–467
Holland JP, Divilov V, Bander NH, Smith-Jones PM, Larson SM, Lewis JS (2010) 89Zr-DFO-J591 for immunoPET imaging of prostate-specific membrane antigen (PSMA) expression in vivo. J Nucl Med 51:1293–1300
Deri MA, Zeglis BM, Francesconi LC, Lewis JS (2013) PET imaging with 89Zr: from radiochemistry to the clinic. Nucl Med Biol 40:3–14
Abou DS, Ku T, Smith-Jones PM (2011) In vivo biodistribution and accumulation of 89 Zr in mice. Nucl Med Biol 38:675–681
Van Dongen GA, Visser GW, Lub-de Hooge MN et al (2007) Immuno-PET: a navigator in monoclonal antibody development and applications. Oncologist 12:1379–1389
Hoppin J, Orcutt KD, Hesterman JY, Silva MD, Cheng D, Lackas C, Rusckowski M (2011) Assessing antibody pharmacokinetics in mice with in vivo imaging. J Pharmacol Exp Ther 337:350–358
Jain M, Venkatraman G, Batra SK (2007) Optimization of radioimmunotherapy of solid tumors: biological impediments and their modulation. Clin Cancer Res 13:1374–1382
Delahunt C, Horning MP, Wilson BK, Proctor JL, Hegg MC (2014) Limitations of haemozoin-based diagnosis of Plasmodium falciparum using dark-field microscopy. Malar J 13:147
Pasternak ND, Dzikowski R (2009) PfEMP1: an antigen that plays a key role in the pathogenicity and immune evasion of the malaria parasite Plasmodium falciparum. Int J Biochem Cell Biol 41:1463–1466
Jauw YW, Menke-van der Houven CW, van Oordt OHS et al (2016) Immuno-positron emission tomography with zirconium-89-labeled monoclonal antibodies in oncology: what can we learn from initial clinical trials? Front Pharmacol 7. https://doi.org/10.3389/fphar.2016.00131
Elsässer-Beile U, Reischl G, Wiehr S et al (2009) PET imaging of prostate cancer xenografts with a highly specific antibody against the prostate-specific membrane antigen. J Nucl Med 50:606–611
Rolle A-M, Hasenberg M, Thornton CR, Solouk-Saran D, Männ L, Weski J, Maurer A, Fischer E, Spycher PR, Schibli R, Boschetti F, Stegemann-Koniszewski S, Bruder D, Severin GW, Autenrieth SE, Krappmann S, Davies G, Pichler BJ, Gunzer M, Wiehr S (2016) ImmunoPET/MR imaging allows specific detection of Aspergillus fumigatus lung infection in vivo. Proc Natl Acad Sci U S A 113:E1026–E1033
Khater N, Kap M, Sayah R, Elbers D, Vriesendorp HM (2017) Radiolabeled immunoglobulin therapy for patients with solid tumors. J Nucl Med Radiat Ther 8(2)
Boyle CC, Paine AJ, Mather SJ (1992) The mechanism of hepatic uptake of a radiolabelled monoclonal antibody. Int J Cancer 50:912–917
Cataldi M, Vigliotti C, Mosca T et al (2017) Emerging role of the spleen in the pharmacokinetics of monoclonal antibodies, nanoparticles and exosomes. Int J Mol Sci 18. https://doi.org/10.3390/ijms18061249
Mazhar F, Haider N (2016) Respiratory manifestation of malaria: An update. Int J Med Res Health Sci 5:59–65
Yip V, Palma E, Tesar DB, Mundo EE, Bumbaca D, Torres EK, Reyes NA, Shen BQ, Fielder PJ, Prabhu S, Khawli LA, Boswell CA (2014) Quantitative cumulative biodistribution of antibodies in mice: effect of modulating binding affinity to the neonatal fc receptor. MAbs 6:689–696
Cor Bester (Preclinical Drug Development Platform North West University) is thanked for assisting with the animal handling and Delene van Wyk (Steve Biko Academic Hospital) for assisting with the scintigraphic imaging.
This work was supported by the Nuclear Technologies in Medicine and the Biosciences Initiative (NTeMBI), a national technology platform developed and managed by the South African Nuclear Energy Corporation (Necsa) and funded by the Department of Science and Technology to JD, TE and JZ. LB received funding from the South African Research Chairs Initiative of the Department of Science and Technology, administered through the South African National Research Foundation (UID84627).
All procedures relating to the [89Zr]IIIB6 study were approved by North West University’s AnimCare Ethical Committee (NWU-00042-12-A5). All procedures involving the [89Zr]h-R3 were approved by the Animal Ethics Committee (AEC) of University of Western Australia (RA/3/100/1236). Ethical approval for the in vitro studies was granted by the University of Pretoria Ethical committee (No.: 120821-077).
Conflict of Interest
The authors declare that they have no conflict of interest.
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
About this article
Cite this article
Duvenhage, J., Ebenhan, T., Garny, S. et al. Molecular Imaging of a Zirconium-89 Labeled Antibody Targeting Plasmodium falciparum–Infected Human Erythrocytes. Mol Imaging Biol 22, 115–123 (2020). https://doi.org/10.1007/s11307-019-01360-3
- Plasmodium falciparum
- Micro-PET/CT imaging