메뉴 건너뛰기




Volumn 342, Issue 6158, 2013, Pages 592-598

Structure-based design of a fusion glycoprotein vaccine for respiratory syncytial virus

Author keywords

[No Author keywords available]

Indexed keywords

GLYCOPROTEIN; MOTAVIZUMAB; PALIVIZUMAB; RESPIRATORY SYNCYTIAL VIRUS VACCINE; VIRUS FUSION PROTEIN;

EID: 84887277978     PISSN: 00368075     EISSN: 10959203     Source Type: Journal    
DOI: 10.1126/science.1243283     Document Type: Article
Times cited : (745)

References (49)
  • 1
    • 84871036375 scopus 로고    scopus 로고
    • R. Lozano et al., Lancet 380, 2095-2128 (2012).
    • (2012) Lancet , vol.380 , pp. 2095-2128
    • Lozano, R.1
  • 3
    • 0031683919 scopus 로고    scopus 로고
    • The IMpact-RSV Study Group
    • The IMpact-RSV Study Group, Pediatrics 102, 531-537 (1998).
    • (1998) Pediatrics , vol.102 , pp. 531-537
  • 7
    • 84878349946 scopus 로고    scopus 로고
    • J. S. McLellan et al., Science 340, 1113-1117 (2013).
    • (2013) Science , vol.340 , pp. 1113-1117
    • McLellan, J.S.1
  • 9
    • 84876735541 scopus 로고    scopus 로고
    • L. J. Anderson et al., Vaccine 31 (suppl. 2), B209-B215 (2013).
    • (2013) Vaccine , vol.31 , Issue.SUPPL. 2
    • Anderson, L.J.1
  • 11
    • 84878345789 scopus 로고    scopus 로고
    • RSV-specific binding molecules and means for producing them
    • Patent Application 12/600,950
    • H. Spits, T. Beaumont, RSV-specific binding molecules and means for producing them. Patent Application 12/600,950 (2010).
    • (2010)
    • Spits, H.1    Beaumont, T.2
  • 12
    • 84878348613 scopus 로고    scopus 로고
    • RSV specific binding molecule
    • Patent Application 12/898,325
    • T. Beaumont, A. Q. Bakker, E. Yasuda, RSV specific binding molecule. Patent Application 12/898,325 (2012).
    • (2012)
    • Beaumont, T.1    Bakker, A.Q.2    Yasuda, E.3
  • 14
    • 33847101745 scopus 로고    scopus 로고
    • T. Zhou et al., Nature 445, 732-737 (2007).
    • (2007) Nature , vol.445 , pp. 732-737
    • Zhou, T.1
  • 16
    • 84876215674 scopus 로고    scopus 로고
    • J. Jardine et al., Science 340, 711-716 (2013).
    • (2013) Science , vol.340 , pp. 711-716
    • Jardine, J.1
  • 23
    • 84887270083 scopus 로고    scopus 로고
    • note
    • Single-letter abbreviations for the amino acid residues are as follows: A, Ala; C, Cys; D, Asp; E, Glu; F, Phe; G, Gly; H, His; I, Ile; K, Lys; L, Leu; M, Met; N, Asn; P, Pro; Q, Gln; R, Arg; S, Ser; T, Thr; V, Val; W, Trp; and Y, Tyr.
  • 24
    • 84855502585 scopus 로고    scopus 로고
    • Information on materials and methods is available on
    • Information on materials and methods is available on Science Online.
    • Science Online
  • 25
    • 84887291765 scopus 로고    scopus 로고
    • note
    • RSV F variants were assessed by transient expression in 96-well format (fig. S4). If supernatants retained reactivity with antibodies motavizumab and D25 after 1 week at 4°C, they were expressed by transient transfection of 1 liter of Expi293F cells, purified by use of appended His-tag and StreptagII, and analyzed by size-exclusion chromatography (fig. S7).
  • 26
    • 84887291963 scopus 로고    scopus 로고
    • note
    • The inability to express potential interchain double-cysteine substitutions, despite reasonable modeling in the mature prefusion F1F2 structure, may indicate that RSV F0 protomers, before cleavage and removal of peptide 27, adopt substantially different interprotomer conformations.
  • 27
    • 84887298275 scopus 로고    scopus 로고
    • note
    • Cavities in the D25-bound RSV F structure were visualized with PyMol using the "Cavities & Pockets Only" option for Surface settings. Amino acid substitutions designed to fill the resulting cavities were identified using the Mutagenesis wizard.
  • 29
    • 84887314828 scopus 로고    scopus 로고
    • note
    • The engineered RSV F variants had Ca-root mean square deviations from the D25-bound conformation of 0.7 to 1.5 Å and from the postfusion conformation of approximately 30 Å.
  • 30
    • 84887268417 scopus 로고    scopus 로고
    • note
    • Crystallized DS retained ∼20% of its D25 recognition relative to crystallized Cav1 (normalized to motavizumab recognition) after 4 months at 20°C, thereby indicating that the crystallized DS was both capable of recognizing D25 as well as converting to a conformation incompatible with D25 binding. By contrast, soluble DS lost all recognition of D25 after 2 months of incubation at 4°C in PBS. We were unable to crystallize DS, which had been heat triggered at 50°C, despite the heat-triggered DS retaining a trimeric state on size exclusion chromatography.
  • 31
    • 84887307458 scopus 로고    scopus 로고
    • note
    • Observing Ala107 rather than Arg109 at the C terminus of F2 is not totally unexpected: Garten and Klenk (48) found that an arginine at the cleavage site of influenza hemagglutinin was trimmed by a cellular carboxypeptidase activity.
  • 32
    • 84887308714 scopus 로고    scopus 로고
    • note
    • Ca-root mean square deviations of 0.86 Å for 447 residues between Cav1 and DS-Cav1 crystal structures obtained from ammonium sulfate; Ca-root mean square deviations of 0.47 Å for 447 residues of DS-Cav1 in cubic lattices.
  • 33
  • 34
    • 84887306542 scopus 로고    scopus 로고
    • note
    • When palivizumab (Synagis) is dosed at a concentration of 15 mg/kg, serum levels at trough are ∼40 mg/ml, which provides protection in infants from severe disease and protection in cotton rats from RSV infection. In our neutralization assay, ∼40 mg/ml of palivizumab yields an EC50 of 100.
  • 35
    • 70349658837 scopus 로고    scopus 로고
    • G. J. Nabel, Science 326, 53-54 (2009).
    • (2009) Science , vol.326 , pp. 53-54
    • Nabel, G.J.1
  • 36
    • 84887285936 scopus 로고    scopus 로고
    • note
    • By contrast, affinity of RSV F variants for D25 antibody did not show correlation with elicitation of protective titers. Yield of RSV F trimers also did not correlate with protective titers.
  • 37
    • 84887281144 scopus 로고    scopus 로고
    • note
    • Flexibility of an antigenic site may increase its immunogenicity by allowing the site to conform to a wider diversity of antibodies. We note in this context that the atomic-mobility factors of antigenic site Ø were among the highest in the RSV F ectodomain.
  • 38
    • 84887312625 scopus 로고    scopus 로고
    • note
    • For the "prefusion" form of RSV F, we used the DS-Cav1 stabilized variant of RSV F.
  • 39
    • 84887310594 scopus 로고    scopus 로고
    • note
    • It should be possible to deconvolute the elicited response, by using structurally defined probes, as shown with D25 and motavizumab-bound RSV F in fig. S12.
  • 41
    • 80054836686 scopus 로고    scopus 로고
    • M. L. Azoitei et al., Science 334, 373-376 (2011).
    • (2011) Science , vol.334 , pp. 373-376
    • Azoitei, M.L.1
  • 46
    • 84887310040 scopus 로고    scopus 로고
    • note
    • Although an epitope-scaffold strategy may not elicit titers as high as a neutralization-sensitive site strategy, for specific antibodies - such as antibody MPE8 (49), which is capable of neutralizing not only RSV but other paramyxoviruses including human metapneumovirus - an epitope-specific strategy may suffice.
  • 47
    • 84887277642 scopus 로고    scopus 로고
    • note
    • The magnitude of antibody activity is a crucial determinant of how well an individual will be protected following vaccination and how long protective responses will be maintained in infants who have passively acquired antibody from their mothers.
  • 49
    • 84884413000 scopus 로고    scopus 로고
    • D. Corti et al., Nature 501, 439-443 (2013).
    • (2013) Nature , vol.501 , pp. 439-443
    • Corti, D.1


* 이 정보는 Elsevier사의 SCOPUS DB에서 KISTI가 분석하여 추출한 것입니다.