ジャーナル: NPJ Vaccines / 年: 2021 タイトル: Immunofocusing and enhancing autologous Tier-2 HIV-1 neutralization by displaying Env trimers on two-component protein nanoparticles. 著者: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / ...著者: Philip J M Brouwer / Aleksandar Antanasijevic / Marlon de Gast / Joel D Allen / Tom P L Bijl / Anila Yasmeen / Rashmi Ravichandran / Judith A Burger / Gabriel Ozorowski / Jonathan L Torres / Celia LaBranche / David C Montefiori / Rajesh P Ringe / Marit J van Gils / John P Moore / Per Johan Klasse / Max Crispin / Neil P King / Andrew B Ward / Rogier W Sanders / 要旨: The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels ...The HIV-1 envelope glycoprotein trimer is poorly immunogenic because it is covered by a dense glycan shield. As a result, recombinant Env glycoproteins generally elicit inadequate antibody levels that neutralize clinically relevant, neutralization-resistant (Tier-2) HIV-1 strains. Multivalent antigen presentation on nanoparticles is an established strategy to increase vaccine-driven immune responses. However, due to nanoparticle instability in vivo, the display of non-native Env structures, and the inaccessibility of many neutralizing antibody (NAb) epitopes, the effects of nanoparticle display are generally modest for Env trimers. Here, we generate two-component self-assembling protein nanoparticles presenting twenty SOSIP trimers of the clade C Tier-2 genotype 16055. We show in a rabbit immunization study that these nanoparticles induce 60-fold higher autologous Tier-2 NAb titers than the corresponding SOSIP trimers. Epitope mapping studies reveal that the presentation of 16055 SOSIP trimers on these nanoparticle focuses antibody responses to an immunodominant apical epitope. Thus, these nanoparticles are a promising platform to improve the immunogenicity of Env trimers with apex-proximate NAb epitopes.
ダウンロード / ファイル: emd_22714.map.gz / 形式: CCP4 / 大きさ: 42.9 MB / タイプ: IMAGE STORED AS FLOATING POINT NUMBER (4 BYTES)
注釈
nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2463
ボクセルのサイズ
X=Y=Z: 1.77 Å
密度
表面レベル
登録者による: 0.015 / ムービー #1: 0.015
最小 - 最大
-0.047056224 - 0.07574413
平均 (標準偏差)
-9.061828e-05 (±0.0033965872)
対称性
空間群: 1
詳細
EMDB XML:
マップ形状
Axis order
X
Y
Z
Origin
0
0
0
サイズ
224
224
224
Spacing
224
224
224
セル
A=B=C: 396.47998 Å α=β=γ: 90.0 °
CCP4マップ ヘッダ情報:
mode
Image stored as Reals
Å/pix. X/Y/Z
1.77
1.77
1.77
M x/y/z
224
224
224
origin x/y/z
0.000
0.000
0.000
length x/y/z
396.480
396.480
396.480
α/β/γ
90.000
90.000
90.000
MAP C/R/S
1
2
3
start NC/NR/NS
0
0
0
NC/NR/NS
224
224
224
D min/max/mean
-0.047
0.076
-0.000
-
添付データ
-
試料の構成要素
-
全体 : nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl...
全体
名称: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2463 (Week 22)
要素
複合体: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2463 (Week 22)
-
超分子 #1: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polycl...
超分子
名称: nsEM map of the 16055 SOSIP HIV Env trimer in complex with polyclonal Fab from rabbit r2463 (Week 22) タイプ: complex / ID: 1 / 親要素: 0 / 含まれる分子: #1
由来(天然)
生物種: synthetic construct (人工物)
組換発現
生物種: Homo sapiens (ヒト) / 組換株: HEK293F / 組換プラスミド: pPPI4
-
実験情報
-
構造解析
手法
ネガティブ染色法
解析
単粒子再構成法
試料の集合状態
particle
-
試料調製
濃度
0.05 mg/mL
緩衝液
pH: 7.4 構成要素:
濃度
名称
式
25.0 mM
Tris-HCl
150.0 mM
sodium chloride
NaCl
詳細: TBS buffer, pH 7.4
染色
タイプ: NEGATIVE / 材質: Uranyl Formate 詳細: Sample diluted to 0.05 mg/mL. 3 uL was applied onto the grid, blotted off, and then stained with 2% uranyl formate for 60 seconds.
16055 SOSIP.v8.3 was complexed with the purified polyclonal Fab overnight. The complex was purified using SEC and concentrated. The complex was then diluted in TBS to 0.05mg/ml to be loaded onto an imaging grid.