Dense Bodies of a gH/gL/UL128/UL130/UL131 Pentamer-Repaired Towne Strain of Human Cytomegalovirus Induce an Enhanced Neutralizing Antibody Response

Lehmann, Caroline; Falk, Jessica; Büscher, Nicole; Penner, Inessa; Zimmermann, Christine; Gogesch, Patricia; Sinzger, Christian; Plachter, Bodo

doi:10.1128/jvi.00931-19

Cited by 16 publications

(41 citation statements)

References 65 publications

(95 reference statements)

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“…Importantly, the DISC vaccine retains the ability to produce viral dense bodies, which is separate from capsid assembly (47). Potentially, dense bodies are important for induction of an overall immune response for both the antibody response to PC and other glycoprotein complexes and the T cell response to pp65 (66).…”

Section: Discussionmentioning

confidence: 99%

Inclusion of the Viral Pentamer Complex in a Vaccine Design Greatly Improves Protection against Congenital Cytomegalovirus in the Guinea Pig Model

Choi

McGregor

2019

J Virol

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A vaccine against congenital cytomegalovirus (cCMV) is a high priority. The guinea pig is a small-animal model for cCMV. A disabled infectious single-cycle (DISC) viral vaccine strain based on a guinea pig cytomegalovirus (GPCMV) capsid mutant was evaluated. A previous version of this vaccine did not express the gH/gL-based pentamer complex (PC) and failed to fully protect against cCMV. The PC is necessary for GPCMV epithelial cell/trophoblast tropism and congenital infection and is a potentially important neutralizing antigen. Here, we show that a second-generation PC-positive (PC+) DISC (DISCII) vaccine induces neutralizing antibodies to the PC and other glycoproteins and a cell-mediated response to pp65 (GP83). Additionally, a CRISPR/Cas9 strategy identified guinea pig platelet-derived growth factor receptor alpha (PDGFRA) to be the receptor for PC-independent infection of fibroblast cells. Importantly, PDGFRA was absent in epithelial and trophoblast cells, which were dependent upon the viral PC for infection. Virus neutralization by DISCII antibodies on epithelial and trophoblast cells was similar to that in sera from wild-type virus-infected animals and dependent in part on PC-specific antibodies. In contrast, sera from PC-negative virus-infected animals poorly neutralized virus on non-fibroblast cells. DISCII-vaccinated animals were protected against congenital infection, in contrast to a nonvaccinated group. The target organs of pups in the vaccine group were negative for wild-type virus, unlike those of pups in the control group, with GPCMV transmission being approximately 80%. Overall, the DISCII vaccine had 97% efficacy against cCMV. The complete protection provided by this PC+ DISC vaccine makes the possibility of the use of this approach against human cCMV attractive. IMPORTANCE Cytomegalovirus (CMV) is a leading cause of congenital disease in newborns, and an effective vaccine remains an elusive goal. The guinea pig is the only small-animal model for cCMV. Guinea pig cytomegalovirus (GPCMV) encodes a glycoprotein pentamer complex (PC) for entry into non-fibroblast cells, including placental trophoblasts, to enable cCMV. As with human cytomegalovirus (HCMV), GPCMV uses a specific cell receptor (PDGFRA) for fibroblast entry, but other receptors are required for non-fibroblast cells. A disabled infectious single-cycle (DISC) GPCMV vaccine strain induced an antibody immune response to the viral pentamer to enhance virus neutralization on non-fibroblast cells, and vaccinated animals were fully protected against cCMV. Inclusion of the PC as part of a vaccine design dramatically improved vaccine efficacy, and this finding underlines the importance of the immune response to the PC in contributing toward protection against cCMV. This vaccine represents an important milestone in the development of a vaccine against cCMV.

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Section: Discussionmentioning

confidence: 99%

Inclusion of the Viral Pentamer Complex in a Vaccine Design Greatly Improves Protection against Congenital Cytomegalovirus in the Guinea Pig Model

Choi

McGregor

2019

J Virol

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“…One vaccine candidate that has been studied in our laboratory and by others is based on subviral particles of HCMV, known as dense bodies (DB) [25,26,27,28,29,30,31,32,33,34,35] (Table 1). DB are synthesized in infected fibroblast cell cultures and are released from these cells at late stages of HCMV replication, concomitant with the release of virions [36,37] (Figure 1).…”

Section: Introductionmentioning

confidence: 99%

Production Strategies for Pentamer-Positive Subviral Dense Bodies as a Safe Human Cytomegalovirus Vaccine

et al. 2019

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Infections with the human cytomegalovirus (HCMV) are associated with severe clinical manifestations in children following prenatal transmission and after viral reactivation in immunosuppressed individuals. The development of an HCMV vaccine has long been requested but there is still no licensed product available. Subviral dense bodies (DB) are immunogenic in pre-clinical models and are thus a promising HCMV vaccine candidate. Recently, we established a virus based on the laboratory strain Towne that synthesizes large numbers of DB containing the pentameric protein complex gH/gL/UL128-131 (Towne-UL130repΔGFP). The work presented here focuses on providing strategies for the production of a safe vaccine based on that strain. A GMP-compliant protocol for DB production was established. Furthermore, the DB producer strain Towne-UL130rep was attenuated by deleting the UL25 open reading frame. Additional genetic modifications aim to abrogate its capacity to replicate in vivo by conditionally expressing pUL51 using the Shield-1/FKBP destabilization system. We further show that the terminase inhibitor letermovir can be used to reduce infectious virus contamination of a DB vaccine by more than two orders of magnitude. Taken together, strategies are provided here that allow for the production of a safe and immunogenic DB vaccine for clinical testing.

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“…This strain synthesized PC-positive DB in infected fibroblasts, which elicited an NAb response higher than that induced by PC-negative DB in both mice and rabbits. The same result was obtained in fibroblast, epithelial, and endothelial cells [136]. On this basis, a GMP-compliant protocol for DB production for use as a vaccine was proposed [137].…”

Section: Multiple Strain Hcmv Infection and Hcmv Vaccinementioning

confidence: 56%

Human Cytomegalovirus Congenital (cCMV) Infection Following Primary and Nonprimary Maternal Infection: Perspectives of Prevention through Vaccine Development

Gerna

Lilleri

2020

Vaccines

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Congenital cytomegalovirus (cCMV) might occur as a result of the human cytomegalovirus (HCMV) primary (PI) or nonprimary infection (NPI) in pregnant women. Immune correlates of protection against cCMV have been partly identified only for PI. Following either PI or NPI, HCMV strains undergo latency. From a diagnostic standpoint, while the serological criteria for the diagnosis of PI are well-established, those for the diagnosis of NPI are still incomplete. Thus far, a recombinant gB subunit vaccine has provided the best results in terms of partial protection. This partial efficacy was hypothetically attributed to the post-fusion instead of the pre-fusion conformation of the gB present in the vaccine. Future efforts should be addressed to verify whether a new recombinant gB pre-fusion vaccine would provide better results in terms of prevention of both PI and NPI. It is still a matter of debate whether human hyperimmune globulin are able to protect from HCMV vertical transmission. In conclusion, the development of an HCMV vaccine that would prevent a significant portion of PI would be a major step forward in the development of a vaccine for both PI and NPI. rate to the fetus in pregnancy during PI of seronegative women might occur in 30-40% of cases [1]. Recent studies have reported a similar trend [2][3][4].(b) Immune response. Both innate and adaptive (antibody and T-cell) immune responses are involved in the control of the HCMV infection. Within the innate response, the protective role of: (i) NK cells and, particularly, CD57 + NKG2C bright cells [5], (ii) antibody-dependent (AD) cellular mechanisms [6,7], and (iii) γ/δ T-cells and, particularly, Vδ2γ/δ T-cells [8,9], against HCMV transmission to the fetus, has not been fully investigated. Thus, at the moment, there are no innate immune correlates that distinguish between transmitting (T) and non-transmitting (NT) women [9].Both neutralizing antibodies (NAb) and non-neutralizing binding antibodies have been believed to exert a protective effect against HCMV transmission to the fetus in seronegative pregnant women [10,11]. However, a more recent randomized study did not confirm the protective effect of commercial human immunoglobulin (HIG) preparations in the prevention of cCMV infections, in comparison to non-treated controls [2]. At the moment, the protective role of HIG in the prevention of cCMV awaits confirmation from more extended controlled studies.Up until 2004, three HCMV glycoprotein complexes (gCs) were known to be the major targets of the NAb response-(i) gB (gCI), homotrimer coded by UL55; (ii) gM/gN complex (gCII), consisting of UL100-coded gM and UL73-coded gN; and (iii) gH/gL/gO complex (gCIII), consisting of UL75-coded gH, UL115-coded gL, and UL74-coded gO. In 2004-2005 the UL128-130-131 locus (referred to as UL128L) was found to be indispensable for infections of endothelial [12] and epithelial [13] cells. Subsequently, UL128L gene products were found to be complexed with gH/gL to form the pentameter complex (PC) gH/gL/pUL128L [14]. Then in 2010,...

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Dense Bodies of a gH/gL/UL128/UL130/UL131 Pentamer-Repaired Towne Strain of Human Cytomegalovirus Induce an Enhanced Neutralizing Antibody Response

Cited by 16 publications

References 65 publications

Inclusion of the Viral Pentamer Complex in a Vaccine Design Greatly Improves Protection against Congenital Cytomegalovirus in the Guinea Pig Model

Inclusion of the Viral Pentamer Complex in a Vaccine Design Greatly Improves Protection against Congenital Cytomegalovirus in the Guinea Pig Model

Production Strategies for Pentamer-Positive Subviral Dense Bodies as a Safe Human Cytomegalovirus Vaccine

Human Cytomegalovirus Congenital (cCMV) Infection Following Primary and Nonprimary Maternal Infection: Perspectives of Prevention through Vaccine Development

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