Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.
Rescue of HIV-1 broad neutralizing antibody-expressing B cells in 2F5 VH x VL knockin mice reveals multiple tolerance controls.
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在2F5 VH X VL敲击蛋白小鼠中，HIV-1宽大中和表达抗体的B细胞揭示了多种耐受性控制。
DOI：
10.4049/jimmunol.1101633
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发表时间：
2011-10-01
期刊：
影响因子：
--
通讯作者：
Haynes BF
中科院分区：
文献类型：
--
作者：
Verkoczy L;Chen Y;Bouton-Verville H;Zhang J;Diaz M;Hutchinson J;Ouyang YB;Alam SM;Holl TM;Hwang KK;Kelsoe G;Haynes BF
HIV-1广谱中和抗体（bnAb）2F 5已被证明在体外具有多聚/自身反应性，我们之前证明，其VDJ重排的单独靶向表达足以在2F 5 VH敲入（KI）小鼠中引发深刻的B细胞发育阻滞，这与表达2F 5 H链的B细胞的中心缺失一致。在此，我们产生了表达整个2F 5 bnAb特异性的品系，2F 5 VHxVL KI小鼠，并且发现了比在2F 5 VH KI品系中观察到的更高程度的耐受性对照。尽管2F 5 VHxVL KI动物的B细胞发育严重受损，但我们证明了在IL-7/BAFF中培养时其B细胞的拯救。有趣的是，即使在这些条件下，大多数拯救的B细胞杂交瘤产生的mAb由于2F 5 L链的编辑而缺乏HIV-1包膜（Env）反应性，并且大多数拯救的B细胞保留了无反应性表型。因此，当避免克隆缺失时，κ编辑和无反应性是防止未成熟/过渡B细胞表达2F 5 VH/VL的额外保障措施。重要的是，7%的拯救的B细胞保留2F 5 VH/VL表达，并分泌具有HIV-1中和活性的Env特异性mAb。这种“部分”拯救在体内得到进一步证实，如2F 5 VHxVL KI × Eμ-bcl 2 tg小鼠中大多数拯救的B细胞的无反应性表型以及Env特异性B细胞和血清Ig的显著（但适度）富集所反映的。本研究中拯救的产生2F 5 mAb的B细胞克隆是体内衍生的骨髓前体特异性HIV-1 bnAb的第一个例子，并为旨在拯救此类B细胞的策略设计提供了起点。
The HIV-1 broad neutralizing antibody (bnAb) 2F5 has been shown to be poly/self-reactive in vitro, and we previously demonstrated that targeted expression of its VDJ rearrangement alone was sufficient to trigger a profound B cell developmental blockade in 2F5 VH knockin (KI) mice, consistent with central deletion of 2F5 H chain-expressing B cells. Here, we generate a strain expressing the entire 2F5 bnAb specificity, 2F5 VHxVL KI mice, and find an even higher degree of tolerance control than observed in the 2F5 VH KI strain. Although B-cell development was severely impaired in 2F5 VHxVL KI animals, we demonstrate rescue of their B-cells when cultured in IL-7/BAFF. Intriguingly, even under these conditions, most rescued B-cell hybridomas produced mAbs that lacked HIV-1 Envelope (Env) reactivity due to editing of the 2F5 L chain, and the majority of rescued B-cells retained an anergic phenotype. Thus, when clonal deletion is circumvented, κ editing and anergy are additional safeguards preventing 2F5 VH/VL expression by immature/transitional B-cells. Importantly, 7% of rescued B-cells retained 2F5 VH/VL-expression and secreted Env-specific mAbs with HIV-1 neutralizing activity. This “partial” rescue was further corroborated in vivo, as reflected by the anergic phenotype of most rescued B-cells in 2F5 VHxVL KI × Eμ-bcl2 tg mice, and significant (yet modest) enrichment of Env-specific B-cells and serum Igs. The rescued 2F5 mAb-producing B-cell clones in this study are the first examples of in vivo-derived bone marrow precursors specifying HIV-1 bnAbs, and provide a starting point for design of strategies aimed at rescuing such B-cells.
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DOI：
10.1084/jem.186.9.1513
发表时间：
1997-11-03
期刊：
The Journal of experimental medicine
影响因子：
--
作者：
Lang J;Arnold B;Hammerling G;Harris AW;Korsmeyer S;Russell D;Strasser A;Nemazee D
通讯作者：
Nemazee D
影响因子：
56.9
作者：
Haynes, BF;Fleming, J;Alam, SM
通讯作者：
Alam, SM
影响因子：
64.5
作者：
HARTLEY, SB;COOKE, MP;GOODNOW, CC
通讯作者：
GOODNOW, CC
影响因子：
15.3
作者：
RUBIN, RL;BALDERAS, RS;THEOFILOPOULOS, AN
通讯作者：
THEOFILOPOULOS, AN
影响因子：
--
作者：
Jiang, Chuancang;Zhao, Ming-Lang;Scearce, Richard M.;Diaz, Marilyn
通讯作者：
Diaz, Marilyn