Liu Hui, Gemmell Lorraine, Lin Rui, Zuo Fengrong, Balfour Henry H, Woo Jennifer C, Hayes Gregory M
AstraZeneca, South San Francisco, CA 94080, USA.
University of Minnesota Medical School, Minneapolis, MN 55455, USA.
Mediterr J Hematol Infect Dis. 2020 Mar 1;12(1):e2020016. doi: 10.4084/MJHID.2020.016. eCollection 2020.
No licensed vaccine is available for prevention of EBV-associated diseases, and robust, high-throughput bioanalytical assays are needed to evaluate immunogenicity of gp350 subunit-based candidate EBV vaccines. Here we have developed an improved EBV-GFP based neutralization assay for such a vaccine's pre-clinical and clinical validation to measure EBV specific neutralizing antibodies in human donors. The supplementation of guinea pig complement of our previously published high-throughput EBV-GFP fluorescent focus (FFA)-based neutralization assay allowed the detection of complement-dependent neutralizing antibodies using a panel of heat-inactivated healthy human sera. Anti-gp350 antibody titers, which were evaluated using a previously optimized anti-gp350 IgG ELISA assay, were moderately correlated to the FFA-based neutralization titers. Overall, this improved high-throughput neutralization assay is capable of characterizing the serologic neutralizing antibody response to natural EBV infection, with applications in evaluating EBV antibody status in epidemiologic studies and immunogenicity of candidate gp350-subunit EBV vaccines in clinical studies.
目前尚无用于预防EBV相关疾病的许可疫苗，因此需要强大的高通量生物分析检测方法来评估基于gp350亚基的候选EBV疫苗的免疫原性。在此，我们开发了一种改进的基于EBV-GFP的中和检测方法，用于此类疫苗的临床前和临床验证，以测量人类供体中EBV特异性中和抗体。在我们之前发表的基于高通量EBV-GFP荧光灶（FFA）的中和检测方法中补充豚鼠补体，使得能够使用一组热灭活的健康人血清检测补体依赖性中和抗体。使用先前优化的抗gp350 IgG ELISA检测方法评估的抗gp350抗体滴度与基于FFA的中和滴度呈中度相关。总体而言，这种改进的高通量中和检测方法能够表征对自然EBV感染的血清中和抗体反应，可应用于在流行病学研究中评估EBV抗体状态以及在临床研究中评估候选gp350亚基EBV疫苗的免疫原性。
