Assessment of precision in growth inhibition assay (GIA) using human anti-PfRH5 antibodies.
Assessment of precision in growth inhibition assay (GIA) using human anti-PfRH5 antibodies.
复制标题
DOI：
10.1186/s12936-023-04591-6
复制
发表时间：
2023-05-19
期刊：
影响因子：
3
通讯作者：
Long, Carole A.
中科院分区：
文献类型：
--
作者：
Miura, Kazutoyo;Diouf, Ababacar;Fay, Michael P.;Barrett, Jordan R.;Payne, Ruth O.;Olotu, Ally I.;Minassian, Angela M.;Silk, Sarah E.;Draper, Simon J.;Long, Carole A.
对于血液阶段疟疾疫苗的开发，体外生长抑制试验（GIA）已被广泛用于评估疫苗诱导的抗体（Ab）的功能，并且恶性疟原虫网织红细胞结合蛋白同源物5（RH 5）是主要的血液阶段抗原。然而，GIA读数和EoA来源的精密度（也称为“测定误差（EoA）"）尚未得到系统评价。在主GIA实验中，使用从4个不同供体采集的红细胞（RBC）制备4种不同的恶性疟原虫3D 7寄生虫培养物。对于每种培养物，通过GIA在两种浓度下在三个不同的日期（168个数据点）检测7种不同的抗RH 5 Ab（单克隆或多克隆Ab）。为了评价EoA的来源（GIA中的%抑制（%GIA）），进行了线性模型拟合，包括供体（RBC来源）和GIA日期作为自变量。此外，在临床GIA实验中检测了180种人抗RH 5多克隆Ab，其中使用不同RBC在至少3个独立GIA中以多种浓度检测每种Ab（5，093个数据点）。估计了%GIA和GIA 50（产生50%GIA的抗体浓度）读数的标准偏差（sd），以及重复测定对这些读数的95%置信区间（95%CI）的影响。主GIA实验表明，红细胞供体效应远大于日效应，临床GIA实验也观察到明显的供体效应。%GIA和对数转换的GIA 50数据均合理拟合恒定sd模型，%GIA和对数转换的GIA 50测量值的sd分别计算为7.54和0.206。与单次测定相比，取三次重复测定（使用三种不同的RBC）的平均值使%GIA或GIA 50测量值的95%CI宽度减少约一半。至少对于本研究中评价的RH 5 Ab，GIA中的RBC供体效应（同一天的供体间差异）远大于日效应（使用同一供体RBC的日间差异）;因此，未来的GIA研究应考虑供体效应。此外，此处显示的%GIA和GIA 50的95%CI有助于比较不同样本/组/研究的GIA结果;因此，本研究支持未来的疟疾血液阶段疫苗开发。在线版本包含补充材料，可通过10.1186/s12936-023-04591-6获得。
For blood-stage malaria vaccine development, the in vitro growth inhibition assay (GIA) has been widely used to evaluate functionality of vaccine-induced antibodies (Ab), and Plasmodium falciparum reticulocyte-binding protein homolog 5 (RH5) is a leading blood-stage antigen. However, precision, also called “error of assay (EoA)”, in GIA readouts and the source of EoA has not been evaluated systematically. In the Main GIA experiment, 4 different cultures of P. falciparum 3D7 parasites were prepared with red blood cells (RBC) collected from 4 different donors. For each culture, 7 different anti-RH5 Ab (either monoclonal or polyclonal Ab) were tested by GIA at two concentrations on three different days (168 data points). To evaluate sources of EoA in % inhibition in GIA (%GIA), a linear model fit was conducted including donor (source of RBC) and day of GIA as independent variables. In addition, 180 human anti-RH5 polyclonal Ab were tested in a Clinical GIA experiment, where each Ab was tested at multiple concentrations in at least 3 independent GIAs using different RBCs (5,093 data points). The standard deviation (sd) in %GIA and in GIA50 (Ab concentration that gave 50%GIA) readouts, and impact of repeat assays on 95% confidence interval (95%CI) of these readouts was estimated. The Main GIA experiment revealed that the RBC donor effect was much larger than the day effect, and an obvious donor effect was also observed in the Clinical GIA experiment. Both %GIA and log-transformed GIA50 data reasonably fit a constant sd model, and sd of %GIA and log-transformed GIA50 measurements were calculated as 7.54 and 0.206, respectively. Taking the average of three repeat assays (using three different RBCs) reduces the 95%CI width in %GIA or in GIA50 measurements by ~ half compared to a single assay. The RBC donor effect (donor-to-donor variance on the same day) in GIA was much bigger than the day effect (day-to-day variance using the same donor’s RBC) at least for the RH5 Ab evaluated in this study; thus, future GIA studies should consider the donor effect. In addition, the 95%CI for %GIA and GIA50 shown here help when comparing GIA results from different samples/groups/studies; therefore, this study supports future malaria blood-stage vaccine development. The online version contains supplementary material available at 10.1186/s12936-023-04591-6.
登录
查看更多内容
影响因子：
7.7
作者：
Ebel ER;Kuypers FA;Lin C;Petrov DA;Egan ES
通讯作者：
Egan ES
影响因子：
5.5
作者：
Malkin, Elissa;Hu, Jinhong;Cao, Zhifang
通讯作者：
Cao, Zhifang
PCRCR 复合物对于恶性疟原虫入侵人类红细胞至关重要。
影响因子：
28.3
作者：
通讯作者：
--
DOI：
10.1016/j.xcrm.2021.100326
发表时间：
2021-07-20
期刊：
Cell reports. Medicine
影响因子：
--
作者：
Willcox AC;Huber AS;Diouf A;Barrett JR;Silk SE;Pulido D;King LDW;Alanine DGW;Minassian AM;Diakite M;Draper SJ;Long CA;Miura K
通讯作者：
Miura K
影响因子：
8
作者：
Payne, Ruth O.;Silk, Sarah E.;Draper, Simon J.
通讯作者：
Draper, Simon J.