Division of Viral Products, Center for Biologics Evaluation and Research (CBER), Food and Drug Administration (FDA), Silver Spring, MD, USA.
Division of Viral Products, Center for Biologics Evaluation and Research (CBER), Food and Drug Administration (FDA), Silver Spring, MD, USA.
Antiviral Res. 2017 Aug;144:8-20. doi: 10.1016/j.antiviral.2017.05.002. Epub 2017 May 8.
Bioluminescence imaging (BLI) was used to follow dissemination of recombinant vaccinia virus (VACV) expressing luciferase (IHD-J-Luc) in BALB/c nu/nu mice treated post-challenge with monoclonal antibodies (MAbs) against L1 and B5 VACV proteins in a model of Progressive Vaccinia (PV). Areas Under the flux Curve (AUC) were calculated for viral loads in multiple organs in individual mice. Following scarification with 10 pfu, IHD-J-Luc VACV undergoes fast replication at the injection site and disseminates rapidly to the inguinal lymph nodes followed by spleen, liver, and axillary lymph nodes within 2-3 days and before primary lesions are visible at the site of scarification. Extension of survival in nude mice treated with a combination of anti-B5 and anti-L1 MAbs 24 h post challenge correlated with a significant reduction in viral load at the site of scarification and delayed systemic dissemination. Nude mice reconstituted with 10 T cells prior to challenge with IHD-J-Luc, and treated with MAbs post-challenge, survived infection, cleared the virus from all organs and scarification site, and developed anti-VACV IgG and VACV-specific polyfunctional CD8 T cells that co-expressed the degranulation marker CD107a, and IFNγ and TNFα cytokines. All T cell reconstituted mice survived intranasal re-challenge with IHD-J-Luc (10 pfu) two months after the primary infection. Thus, using BLI to monitor VACV replication in a PV model, we showed that anti-VACV MAbs administered post challenge extended survival of nude mice and protected T cell reconstituted nude mice from lethality by reducing replication at the site of scarification and systemic dissemination of VACV.
生物发光成像（BLI）用于监测表达荧光素酶的重组痘苗病毒（VACV）（IHD-J-Luc）在接种后用针对 L1 和 B5 VACV 蛋白的单克隆抗体（MAb）治疗的 BALB/c nu/nu 小鼠中的播散，以建立进行性痘苗（PV）模型。在单个小鼠的多个器官中，计算病毒载量的通量曲线下面积（AUC）。在接种 10 pfu 后，IHD-J-Luc VACV 在注射部位迅速复制，并在 2-3 天内迅速播散到腹股沟淋巴结，随后播散到脾、肝和腋窝淋巴结，然后在接种部位的原发性病变可见之前。在挑战后 24 小时用抗 B5 和抗 L1 MAb 联合治疗的裸鼠中的生存时间延长与接种部位病毒载量的显著降低以及系统传播的延迟相关。在挑战前用 10 个 T 细胞重建的裸鼠，并用 MAb 治疗后，感染后存活，从所有器官和接种部位清除病毒，并产生抗 VACV IgG 和 VACV 特异性多功能 CD8 T 细胞，其共同表达脱颗粒标记物 CD107a，以及 IFNγ 和 TNFα 细胞因子。所有 T 细胞重建的小鼠在原发性感染两个月后用 IHD-J-Luc（10 pfu）进行鼻内再挑战后均存活。因此，我们使用 BLI 监测 PV 模型中的 VACV 复制，表明在挑战后给予抗 VACV MAb 可延长裸鼠的生存时间，并通过减少接种部位的病毒复制和 VACV 的全身传播来保护 T 细胞重建的裸鼠免于致死性。
