International AIDS Vaccine Initiative Human Immunology Laboratory, Imperial College, London, United Kingdom.
PLoS One. 2010 Dec 14;5(12):e14330. doi: 10.1371/journal.pone.0014330.
The Comprehensive T Cell Vaccine Immune Monitoring Consortium (CTC-VIMC) was created to provide standardized immunogenicity monitoring services for HIV vaccine trials. The ex vivo interferon-gamma (IFN-γ) ELISpot is used extensively as a primary immunogenicity assay to assess T cell-based vaccine candidates in trials for infectious diseases and cancer. Two independent, GCLP-accredited central laboratories of CTC-VIMC routinely use their own standard operating procedures (SOPs) for ELISpot within two major networks of HIV vaccine trials. Studies are imperatively needed to assess the comparability of ELISpot measurements across laboratories to benefit optimal advancement of vaccine candidates.
We describe an equivalence study of the two independently qualified IFN-g ELISpot SOPs. The study design, data collection and subsequent analysis were managed by independent statisticians to avoid subjectivity. The equivalence of both response rates and positivity calls to a given stimulus was assessed based on pre-specified acceptance criteria derived from a separate pilot study.
Detection of positive responses was found to be equivalent between both laboratories. The 95% C.I. on the difference in response rates, for CMV (-1.5%, 1.5%) and CEF (-0.4%, 7.8%) responses, were both contained in the pre-specified equivalence margin of interval [-15%, 15%]. The lower bound of the 95% C.I. on the proportion of concordant positivity calls for CMV (97.2%) and CEF (89.5%) were both greater than the pre-specified margin of 70%. A third CTC-VIMC central laboratory already using one of the two SOPs also showed comparability when tested in a smaller sub-study.
The described study procedure provides a prototypical example for the comparison of bioanalytical methods in HIV vaccine and other disease fields. This study also provides valuable and unprecedented information for future vaccine candidate evaluations on the comparison and pooling of ELISpot results generated by the CTC-VIMC central core laboratories.
全面 T 细胞疫苗免疫监测联合会（CTC-VIMC）成立的目的是为 HIV 疫苗试验提供标准化的免疫原性监测服务。体外干扰素-γ（IFN-γ）ELISpot 广泛用于作为原发性免疫测定法，以评估传染病和癌症临床试验中的基于 T 细胞的候选疫苗。CTC-VIMC 的两个独立的、符合 GCLP 标准的中心实验室通常在两个主要的 HIV 疫苗试验网络中使用自己的 ELISpot 标准操作程序（SOP）。迫切需要进行研究，以评估实验室之间的 ELISpot 测量值的可比性，从而使候选疫苗的最佳进展受益。
我们描述了两种独立资格 IFN-γ ELISpot SOP 的等效性研究。研究设计、数据收集和随后的分析由独立的统计学家管理，以避免主观性。根据来自单独的初步研究得出的预定接受标准，评估了两种反应率和阳性刺激的阳性率的等效性。
发现两种实验室之间的阳性反应检测结果等效。CMV（-1.5%，1.5%）和 CEF（-0.4%，7.8%）反应的反应率差异的 95%置信区间均包含在预定的等效区间[-15%，15%]内。CMV（97.2%）和 CEF（89.5%）阳性率的一致性比例的 95%置信区间下限均大于预定的 70%界限。已经使用两个 SOP 之一的第三个 CTC-VIMC 中心实验室在一项较小的子研究中也显示出可比性。
描述的研究程序为 HIV 疫苗和其他疾病领域的生物分析方法的比较提供了原型示例。该研究还为未来的疫苗候选物评估提供了有价值的、前所未有的信息，即比较和汇总 CTC-VIMC 中心核心实验室生成的 ELISpot 结果。
