Comparison of antibodies directed against human respiratory syncytial virus antigens present in two commercial preparations of human immunoglobulins with different neutralizing activities

Sastre, Patricia; Melero, José A.; Garcı́a-Barreno, Blanca; Palomo, Concepción

doi:10.1016/j.vaccine.2004.06.023

Cited by 11 publications

(12 citation statements)

References 38 publications

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“…These antibodies account for most of the neutralizing activity found in human Ig preparations (not only Respigam but also another Ig preparation, Flebogamma; ref. 24) and in sera of rabbits inoculated with Vac/Fc. These antibodies, however, were absent from sera of rabbits inoculated with Vac/ F TM -, illustrating the importance of the F protein structure presented to the immune system for the antibody outcome.…”

Section: Discussionmentioning

confidence: 99%

Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention

Magro

Más

Chappell

et al. 2012

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

231

212

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Human respiratory syncytial virus (hRSV) is the most important viral agent of pediatric respiratory infections worldwide. The only specific treatment available today is a humanized monoclonal antibody (Palivizumab) directed against the F glycoprotein, administered prophylactically to children at very high risk of severe hRSV infections. Palivizumab, as most anti-F antibodies so far described, recognizes an epitope that is shared by the two conformations in which hRSV_F can fold, the metastable prefusion form and the highly stable postfusion conformation. We now describe a unique class of antibodies specific for the prefusion form of this protein that account for most of the neutralizing activity of either a rabbit serum raised against a vaccinia virus recombinant expressing hRSV_F or a human Ig preparation (Respigam), which was used for prophylaxis before Palivizumab. These antibodies therefore offer unique possibilities for immune intervention against hRSV, and their production should be assessed in trials of hRSV vaccines.

show abstract

Section: Discussionmentioning

confidence: 99%

Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention

Magro

Más

Chappell

et al. 2012

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

231

212

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“…The purification of soluble forms of the HRSV F (F TM À) and G (Gs) surface glycoproteins and their covalent binding to Sepharose beads for the preparation of affinity columns have been reported [Sastre et al, 2004]. The efficiency of these columns for depleting a commercial immunoglobulin preparation (RespiGam) of anti-F and anti-G antibodies was evaluated by titrating by ELISA the antibodies that remained in the material not bound to the beads after affinity chromatography.…”

Section: Depletion Of Anti-hrsv Glycoprotein Antibodies By Affinity Cmentioning

confidence: 99%

“…Although murine monoclonal antibodies have provided valuable information about the antigenic structure of HRSV F [Beeler and van Wyke Coelingh, 1989;Arbiza et al, 1992;López et al, 1998] and G glycoproteins [Anderson et al, 1985;Martínez et al, 1997;Melero et al, 1997], human antibodies may recognize epitopes outside the antigenic areas identified by murine antibodies [Scopes et al, 1990;Sastre et al, 2004]. Thus, we have started the characterization of anti-HRSV antibodies present in two commercial preparations of human immunoglobulins that differ in their titres of neutralizing antibodies [Sastre et al, 2004].…”

Section: Introductionmentioning

confidence: 99%

“…Thus, we have started the characterization of anti-HRSV antibodies present in two commercial preparations of human immunoglobulins that differ in their titres of neutralizing antibodies [Sastre et al, 2004]. Antibodies were purified from the immunoglobulin preparations by affinity chromatography using soluble forms of the F (F TM À) and G (Gs) glycoproteins linked covalently to Sepharose beads.…”

Section: Introductionmentioning

confidence: 99%

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Comparison of affinity chromatography and adsorption to vaccinia virus recombinant infected cells for depletion of antibodies directed against respiratory syncytial virus glycoproteins present in a human immunoglobulin preparation

Sastre

Melero

Garcı́a-Barreno

et al. 2005

Journal of Medical Virology

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Antibodies directed against human respiratory syncytial virus (HRSV) glycoproteins were depleted from a commercial immunoglobulin preparation (RespiGam) by two different methods. The first method consisted of repeated adsorption of RespiGam to Sepharose beads with covalently bound soluble forms of the two major viral glycoproteins (F or G). The second method consisted of adsorption of immunoglobulins to live cells expressing F or G glycoproteins on their surfaces after infection with vaccinia virus recombinants. While the first method removed efficiently antibodies that reacted with F and/or G glycoproteins by ELISA, it was inefficient in the elimination of anti-HRSV neutralizing antibodies. In contrast, the second method removed efficiently anti-HRSV antibodies that both reacted by ELISA and neutralized virus infectivity. These results confirm that human neutralizing antibodies are directed exclusively against HRSV F and G glycoproteins, and, they raise the possibility that F and G glycoproteins inserted into cell membranes differ antigenically from their soluble forms linked covalently to Sepharose beads.

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“…Some antibodies specifically recognize the pre-F conformation (32), others are directed against the post-F-specific 6HB structure (33), and the others bind both pre-F and post-F structures, such as palivizumab (34)(35)(36)(37). Importantly, data suggested that a hRSV subunit vaccine based on pre-F triggers a broad protective antibody response, as it has been shown that most of the neutralizing antibodies produced during natural infection are directed against the pre-F conformation (24,38,39). In addition, it was reported previously that monoclonal neutralizing antibodies directed against the pre-F state can prevent the conformational changes leading to the post-F state (32), thereby inhibiting virus-cell fusion.…”

mentioning

confidence: 99%

Characterization of Pre-F-GCN4t, a Modified Human Respiratory Syncytial Virus Fusion Protein Stabilized in a Noncleaved Prefusion Conformation

Blais¹,

Gagné²,

Hamuro³

et al. 2017

J Virol

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The human respiratory syncytial virus (hRSV) fusion (F) protein is considered a major target of the neutralizing antibody response to hRSV. This glycoprotein undergoes a major structural shift from the prefusion (pre-F) to the postfusion (post-F) state at the time of virus-host cell membrane fusion. Recent evidences suggest that the pre-F state is a superior target for neutralizing antibodies compared to the post-F state. Therefore, for vaccine purposes, we have designed and characterized a recombinant hRSV F protein, called Pre-F-GCN4t, stabilized in a pre-F conformation. To show that Pre-F-GCN4t does not switch to a post-F conformation, it was compared with a recombinant post-F molecule, called Post-F-XC. Pre-F-GCN4t was glycosylated and trimeric and displayed a conformational stability different from that of Post-F-XC, as shown by chemical denaturation. Electron microscopy analysis suggested that Pre-F-GCN4t adopts a lollipop-like structure. In contrast, Post-F-XC had a typical elongated conical shape. Hydrogen/deuterium exchange mass spectrometry demonstrated that the two molecules had common rigid folding core and dynamic regions and provided structural insight for their biophysical and biochemical properties and reactivity. Pre-F-GCN4t was shown to deplete hRSV-neutralizing antibodies from human serum more efficiently than Post-F-XC. Importantly, Pre-F-GCN4t was also shown to bind D25, a highly potent monoclonal antibody specific for the pre-F conformation. In conclusion, this construct presents several pre-F characteristics, does not switch to the post-F conformation, and presents antigenic features required for a protective neutralizing antibody response. Therefore, Pre-F-GCN4t can be considered a promising candidate vaccine antigen. IMPORTANCE Human respiratory syncytial virus (RSV) is a global leading cause of infant mortality and adult morbidity. The development of a safe and efficacious RSV vaccine remains an important goal. The RSV class I fusion (F) glycoprotein is considered one of the most promising vaccine candidates, and recent evidences suggest that the prefusion (pre-F) state is a superior target for neutralizing antibodies. Our study presents the physicochemical characterization of Pre-F-GCN4t, a molecule designed to be stabilized in the pre-F conformation. To confirm its pre-F conformation, Pre-F-GCN4t was analyzed in parallel with Post-F-XC, a molecule in the post-F conformation. Our results show that Pre-F-GCN4t presents characteristics of a stabilized pre-F conformation and support its use as an RSV vaccine antigen. Such an antigen may represent a significant advance in the development of an RSV vaccine.

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Comparison of antibodies directed against human respiratory syncytial virus antigens present in two commercial preparations of human immunoglobulins with different neutralizing activities

Cited by 11 publications

References 38 publications

Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention

Neutralizing antibodies against the preactive form of respiratory syncytial virus fusion protein offer unique possibilities for clinical intervention

Comparison of affinity chromatography and adsorption to vaccinia virus recombinant infected cells for depletion of antibodies directed against respiratory syncytial virus glycoproteins present in a human immunoglobulin preparation

Characterization of Pre-F-GCN4t, a Modified Human Respiratory Syncytial Virus Fusion Protein Stabilized in a Noncleaved Prefusion Conformation

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