Isolation of Human Monoclonal Antibodies That Potently Neutralize Human Cytomegalovirus Infection by Targeting Different Epitopes on the gH/gL/UL128-131A Complex

Macagno, Annalisa; Bernasconi, Nadia L.; Vanzetta, Fabrizia; Dander, Erica; Sarasini, Antonella; Revello, Maria Grazia; Gerna, Giuseppe; Sallusto, Federica; Lanzavecchia, Antonio

doi:10.1128/jvi.01809-09

Cited by 303 publications

(391 citation statements)

References 68 publications

(87 reference statements)

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“…Instead, antibodies binding to both gH/gL and Pentamer have been shown to neutralize HCMV infection in all cell types. However, in epithelial and endothelial cells, antibodies specific to the ULs are much more potent (25,26). Our structural data help to rationalize these data and to identify the relative location of two Pentamer regions that are targeted by these different classes of neutralizing antibodies.…”

Section: Discussionmentioning

confidence: 97%

“…Highly potent HCMV-neutralizing monoclonal antibodies that bind the Pentamer, but not gH/gL, have been isolated from the memory B-cell repertoire of HCMV immune donors (25,26). These antibodies were capable of potently neutralizing HCMV infection of epithelial and endothelial cells, but not fibroblasts.…”

Section: Discussionmentioning

confidence: 99%

“…These antibodies were capable of neutralizing HCMV infection of epithelial/endothelial cells, but not fibroblasts (25,26). In addition, several studies have demonstrated that the Pentamer is the main target of the neutralizing humoral response to HCMV infection in epithelial/endothelial cells (27)(28)(29).…”

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confidence: 99%

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Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes

Ciferri

Chandramouli

Donnarumma

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

136

189

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Human cytomegalovirus (HCMV) is a major cause of morbidity and mortality in transplant patients and the leading viral cause of birth defects after congenital infection. The glycoprotein complexes gH/gL/gO and gH/gL/UL128/UL130/UL131A (Pentamer) are key targets of the human humoral response against HCMV and are required for HCMV entry into fibroblasts and endothelial/epithelial cells, respectively. We expressed and characterized soluble forms of gH/gL, gH/gL/gO, and Pentamer. Mass spectrometry and mutagenesis analysis revealed that gL-Cys144 forms disulfide bonds with gO-Cys351 in gH/gL/gO and with UL128-Cys162 in the Pentamer. Notably, Pentamer harboring the UL128-Cys162Ser/gL-Cys144Ser mutations had impaired syncytia formation and reduced interference of HCMV entry into epithelial cells. Electron microscopy analysis showed that HCMV gH/gL resembles HSV gH/gL and that gO and UL128/UL130/UL131A bind to the same site at the gH/gL N terminus. These data are consistent with gH/gL/gO and Pentamer forming mutually exclusive cell entry complexes and reveal the overall location of gH/gL-, gH/gL/ gO-, and Pentamer-specific neutralizing antibody binding sites. Our results provide, to our knowledge, the first structural view of gH/gL/ gO and Pentamer supporting the development of vaccines and antibody therapeutics against HCMV.human cytomegalovirus | HCMV | Pentamer complex | gH/gL/gO | virus entry

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Section: Discussionmentioning

confidence: 97%

Section: Discussionmentioning

confidence: 99%

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Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes

Ciferri

Chandramouli

Donnarumma

et al. 2015

Proc. Natl. Acad. Sci. U.S.A.

136

189

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“…3,11 For instance, primary HCMV infection results in a potent natural killer (NK) cell response, 7 as well as the generation of humoral immunity, which includes neutralizing antibodies, which are specific for a number of viral proteins. 12,13 In addition to antibody responses, primary HCMV infection also results in the generation of both CD4 1 and CD8 1 T cells specific for a very broad range of viral proteins 3,11,14 at very high frequencies ( Figure 1). It is recognized that primary infection of immunosuppressed individuals, or the immunonaive foetus in utero, leads to extensive viral replication in numerous cell types and, ultimately, end-organ disease which can result in serious morbidity and in some cases mortality.…”

Section: Primary Infection and The Immune Response To Virus In Lytic mentioning

confidence: 99%

The immunology of human cytomegalovirus latency: could latent infection be cleared by novel immunotherapeutic strategies?

et al. 2014

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While the host immune response following primary human cytomegalovirus (HCMV) infection is generally effective at stopping virus replication and dissemination, virus is never cleared by the host and like all herpesviruses, persists for life. At least in part, this persistence is known to be facilitated by the ability of HCMV to establish latency in myeloid cells in which infection is essentially silent with, importantly, a total lack of new virus production. However, although the viral transcription programme during latency is much suppressed, a number of viral genes are expressed during latent infection at the protein level and many of these have been shown to have profound effects on the latent cell and its environment. Intriguingly, many of these latency-associated genes are also expressed during lytic infection. Therefore, why the same potent host immune responses generated during lytic infection to these viral gene products are not recognized during latency, thereby allowing clearance of latently infected cells, is far from clear. Reactivation from latency is also a major cause of HCMV-mediated disease, particularly in the immune compromised and immune naive, and is also likely to be a major source of virus in chronic subclinical HCMV infection which has been suggested to be associated with long-term diseases such as atherosclerosis and some neoplasias. Consequently, understanding latency and why latently infected cells appear to be immunoprivileged is crucial for an understanding of the pathogenesis of HCMV and may help to design strategies to eliminate latent virus reservoirs, at least in certain clinical settings.

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“…Subsequently, it was shown that the UL128, UL130 and UL131 locus (thus referred to as UL128L) gene products formed a pentameric complex with gH/gL (gH/gL/pUL128/pUL130/pUL131), which was required for infection of both endothelial and epithelial cells (Wang & Shenk, 2005;Ryckman et al, 2008;. Multiple antigenic sites of the pentamer have been shown to elicit very potent neutralizing mAbs with high frequency, unlike gB which induces a predominant number of binding but non-neutralizing mAbs (Macagno et al, 2010;Pötzsch et al, 2011;Fouts et al, 2012). In addition, gB-based vaccines have shown only partial protection against vertical HCMV transmission in pregnant women and against HCMV infection in transplanted patients (Pass et al, 2009;Griffiths et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Differential kinetics of human cytomegalovirus load and antibody responses in primary infection of the immunocompetent and immunocompromised host

Gerna

Lilleri²,

Fornara

et al. 2015

Journal of General Virology

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The comparative long-term kinetics of human cytomegalovirus (HCMV) load and HCMV-specific antibody responses in the immunocompetent and immunocompromised solid-organ transplanted host during primary HCMV infection was investigated. In total, 40 immunocompetent subjects and 17 transplanted patients were examined for viral load as well as for IgG antibody responses to HCMV glycoproteins gH/gL/pUL128L, gH/gL and gB, and neutralizing antibodies in ARPE-19 epithelial cells and human fibroblasts. In parallel, the CD4 + and CD8 + HCMV-specific T-cell responses were determined by cytokine flow cytometry. Transplanted patients reached significantly higher viral DNA peaks, which persisted longer than in immunocompetent subjects. The ELISA-IgG responses to the pentamer, gH/gL and gB were significantly higher in primary infections of the immunocompetent until six months after onset, with the two antibody levels then overlapping from six to 12 months. Antibody levels neutralizing infection of epithelial cells were significantly higher in transplanted patients after six months, persisting for up to a year after transplantation. This trend was not observed for antibodies neutralizing infection of human fibroblasts, which showed higher titres in the immunocompetent over the entire one-year followup. In conclusion, in immunocompromised patients the viral load peak was much higher, while the neutralizing antibody response exceeded that detected in the immunocompetent host starting six months after onset of follow-up, often concomitantly with a lack of specific CD4 + T cells. In this setting, the elevated antibody response occurred in the presence of differentiated follicular helper T cells in the blood, which decreased in number as did antibody titres upon reappearance of HCMV-specific CD4 + T cells.

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Isolation of Human Monoclonal Antibodies That Potently Neutralize Human Cytomegalovirus Infection by Targeting Different Epitopes on the gH/gL/UL128-131A Complex

Cited by 303 publications

References 68 publications

Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes

Structural and biochemical studies of HCMV gH/gL/gO and Pentamer reveal mutually exclusive cell entry complexes

The immunology of human cytomegalovirus latency: could latent infection be cleared by novel immunotherapeutic strategies?

Differential kinetics of human cytomegalovirus load and antibody responses in primary infection of the immunocompetent and immunocompromised host

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