Background
Transmission of latent human cytomegalovirus (HCMV) via organ transplantation with
post-transplant viral reactivation is extremely prevalent and results in substantial
adverse impact on outcomes. Therapies targeting the latent reservoir within the allograft
to mitigate viral transmission would represent a major advance. Here, we delivered
an immunotoxin (F49A-FTP) that targets and kills latent HCMV aiming at reducing the
HCMV reservoir from donor lungs using ex-vivo lung perfusion (EVLP).
Methods
HCMV seropositive human lungs were placed on EVLP alone or EVLP + 1mg/L of F49A-FTP
for 6 hours (n = 6, each). CD14+ monocytes isolated from biopsies pre and post EVLP underwent HCMV reactivation
assay designed to evaluate viral reactivation capacity. Off-target effects of F49A-FTP
were studied evaluating cell death markers of CD34+ and CD14+ cells using flow cytometry.
Lung function on EVLP and inflammatory cytokine production were evaluated as safety
endpoints.
Results
We demonstrate that lungs treated ex-vivo with F49A-FTP had a significant reduction
in HCMV reactivation compared to controls, suggesting successful targeting of latent
virus (76% median reduction in F49A-FTP vs 15% increase in controls, p = 0.0087). Furthermore, there was comparable cell death rates of the targeted cells between
both groups, suggesting no off-target effects. Ex-vivo lung function was stable over
6 hours and no differences in key inflammatory cytokines were observed demonstrating
safety of this novel treatment.
Conclusions
Ex-vivo F49A-FTP treatment of human lungs targets and kills latent HCMV, markedly
attenuating HCMV reactivation. This approach demonstrates the first experiments targeting
latent HCMV in a donor organ with promising results towards clinical translation.
KEYWORDS
Abbreviations:
HCMV (Human Cytomegalovirus), EVLP (Ex Vivo Lung Perfusion), FTPs (Fusion Toxin Proteins), IE (Immediate-early HCMV protein), mDCs (Monocyte-derived dendritic cells), HFFs (Human foreskin fibroblasts)To read this article in full you will need to make a payment
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References
- Estimation of the worldwide seroprevalence of cytomegalovirus: a systematic review and meta-analysis.Rev Med Virol. 2019; 29: e2034https://doi.org/10.1002/rmv.2034
- OPTN/SRTR 2017 Annual data report: lung.Am J Transplant. 2019; 19: 404-484https://doi.org/10.1111/ajt.15279
- Cytomegalovirus pneumonitis is a risk for bronchiolitis obliterans syndrome in lung transplantation.Am J Respir Crit Care Med. 2010; 181: 1391-1396https://doi.org/10.1164/rccm.200911-1786OC
- Cytomegalovirus replication within the lung allograft Is associated with bronchiolitis obliterans syndrome: CMV in the allograft Is associated with BOS.Am J Transplant. 2011; 11: 2190-2196https://doi.org/10.1111/j.1600-6143.2011.03663.x
- Cytomegalovirus disease among donor-positive/recipient-negative lung transplant recipients in the era of valganciclovir prophylaxis.J Heart Lung Transplant. 2010; 29: 1014-1020https://doi.org/10.1016/j.healun.2010.04.022
- Human cytomegalovirus encoded homologs of cytokines, chemokines and their receptors: roles in immunomodulation.Viruses. 2012; 4: 2448-2470https://doi.org/10.3390/v4112448
- Selective recognition of the membrane-bound CX 3 C chemokine, fractalkine, by the human cytomegalovirus-encoded broad-spectrum receptor US28.FEBS Lett. 1998; 441: 209-214https://doi.org/10.1016/S0014-5793(98)01551-8
- Latency-associated expression of human cytomegalovirus US28 attenuates cell signaling pathways to maintain latent infection.mBio. 2017; 8mBio.01754-17, e01754-17 (Imperiale MJ, ed)https://doi.org/10.1128/mBio.01754-17
- Targeting the latent cytomegalovirus reservoir with an antiviral fusion toxin protein.Nat Commun. 2017; 8: 14321https://doi.org/10.1038/ncomms14321
- Rationally designed chemokine-based toxin targeting the viral G protein-coupled receptor US28 potently inhibits cytomegalovirus infection in vivo.Proc Natl Acad Sci U S A. 2015; 112: 8427-8432https://doi.org/10.1073/pnas.1509392112
- Normothermic ex vivo lung perfusion in clinical lung transplantation.N Engl J Med. 2011; 364: 1431-1440https://doi.org/10.1056/NEJMoa1014597
- Outcomes after transplantation of lungs preserved for more than 12 h: a retrospective study.Lancet Respir Med. 2017; 5: 119-124https://doi.org/10.1016/S2213-2600(16)30323-X
- Functional repair of human donor lungs by IL-10 gene therapy.Sci Transl Med. 2009; 1: 4ra9https://doi.org/10.1126/scitranslmed.3000266
- Annexin V homodimer protects against ischemia reperfusion–induced acute lung injury in lung transplantation.J Thorac Cardiovasc Surg. 2016; 151: 861-869https://doi.org/10.1016/j.jtcvs.2015.10.112
- Ex vivo perfusion treatment of infection in human donor lungs.Am J Transplant. 2016; 16: 1229-1237https://doi.org/10.1111/ajt.13562
- Inactivating hepatitis C virus in donor lungs using light therapies during normothermic ex vivo lung perfusion.Nat Commun. 2019; 10https://doi.org/10.1038/s41467-018-08261-z
- Prevention of viral transmission during lung transplantation with hepatitis C-viraemic donors: an open-label, single-centre, pilot trial.Lancet Respir Med. Published online October 2019; https://doi.org/10.1016/S2213-2600(19)30268-1
- Ex-vivo delivery of monoclonal antibody (Rituximab) to treat human donor lungs prior to transplantation.EBioMed. 2020; 60102994https://doi.org/10.1016/j.ebiom.2020.102994
- Technique for prolonged normothermic ex vivo lung perfusion.J Heart Lung Transplant. 2008; 27: 1319-1325https://doi.org/10.1016/j.healun.2008.09.003
- Molecular determinants and the regulation of human cytomegalovirus latency and reactivation.Viruses. 2018; 10: 444https://doi.org/10.3390/v10080444
- US28: HCMV's Swiss army knife.Viruses. 2018; 10: 445https://doi.org/10.3390/v10080445
- Biomarkers in acute lung injury: insights into the pathogenesis of acute lung injury.Crit Care Clin. 2011; 27: 355-377https://doi.org/10.1016/j.ccc.2010.12.005
- Cytomegalovirus infections in solid organ transplantation: a review.Infect Chemother. 2013; 45: 260-271https://doi.org/10.3947/ic.2013.45.3.260
- Ganciclovir/valganciclovir prophylaxis decreases cytomegalovirus-related events and bronchiolitis obliterans syndrome after lung transplantation.Clin Infectious Dis. 2008; 46: 831-839
- The third international consensus guidelines on the management of cytomegalovirus in solid-organ transplantation.Transplantation. 2018; 102: 900-931
- The third international consensus guidelines on the management of cytomegalovirus in solid-organ transplantation.Transplantation. 2018; 102: 900-931https://doi.org/10.1097/TP.0000000000002191
- Late-onset Cytomegalovirus (CMV) in lung transplant recipients: can CMV serostatus guide the duration of prophylaxis?: lung transplant cytomegalovirus prophylaxis.Am J Transplant. 2013; 13: 376-382https://doi.org/10.1111/j.1600-6143.2012.04339.x
- Drug-resistant cytomegalovirus infection after lung transplantation: incidence, characteristics, and clinical outcomes.J Heart Lung Transplant. 2019; 38: 1268-1274https://doi.org/10.1016/j.healun.2019.09.005
- Prone positioning during ex vivo lung perfusion influences regional edema accumulation.J Surg Res. 2019; 239: 300-308https://doi.org/10.1016/j.jss.2019.02.003
- Injury-specific ex vivo treatment of the donor lung: pulmonary thrombolysis followed by successful lung transplantation.Am J Respir Crit Care Med. 2013; 188: 878-880https://doi.org/10.1164/rccm.201302-0368LE
- Prevention of transfusion-transmitted cytomegalovirus infection.Transfus Med. Published online 1999; 9: 115-123
- Clinical utility of viral load in management of cytomegalovirus infection after solid organ transplantation.Clin Microbiol Rev. 2013; 26: 703-727https://doi.org/10.1128/CMR.00015-13
- Comparison of two quantitative CMV PCR tests, Cobas Amplicor CMV Monitor and TaqMan assay, and pp65-antigenemia assay in the determination of viral loads from peripheral blood of organ transplant patients.J Clin Virol. 2004; 30: 258-266https://doi.org/10.1016/j.jcv.2003.12.010
- Cytomegalovirus replication kinetics in solid organ transplant recipients managed by preemptive therapy: CMV and preemptive antiviral therapy.Am J of Transplant. 2012; 12: 2457-2464https://doi.org/10.1111/j.1600-6143.2012.04087.x
- The conditions of primary infection define the load of latent viral genome in organs and the risk of recurrent cytomegalovirus disease.J Exp Med. 1994; 179: 185-193https://doi.org/10.1084/jem.179.1.185
- Preemptive CD8 T-cell immunotherapy of acute cytomegalovirus infection prevents lethal disease, limits the burden of latent viral genomes, and reduces the risk of virus recurrence.J Virol. 1998; 72: 1797-1804https://doi.org/10.1128/JVI.72.3.1797-1804.1998
- Human cytomegalovirus US28 ligand binding activity is required for latency in CD34 + hematopoietic progenitor cells and humanized NSG cice.mBio. 2019; 10:e01889-19, /mbio/10/4/mBio.01889-19.atom (Shenk T, ed)https://doi.org/10.1128/mBio.01889-19
- HCMV infection of humanized mice after transplantation of G-CSF–mobilized peripheral blood stem cells from HCMV-seropositive donors.Biol Blood Marrow Transplant. 2014; 20: 132-135https://doi.org/10.1016/j.bbmt.2013.10.019
Article info
Publication history
Published online: October 25, 2021
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