System vaccinology for the evaluation of influenza vaccine safety by multiplex gene detection of novel biomarkers in a preclinical study and batch release test.

Takuo Mizukami,Kazuya Takizawa,Kumiko Araki,Kazunari Yamaguchi,Haruka Momose,Isao Hamaguchi,Keiko Furuhata,Ken J Ishii,Madoka Kuramitsu,Sang-Moo Kang

doi:10.1371/journal.pone.0101835

Abstract

Vaccines are beneficial and universal tools to prevent infectious disease. Thus, safety of vaccines is strictly evaluated in the preclinical phase of trials and every vaccine batch must be tested by the National Control Laboratories according to the guidelines published by each country. Despite many vaccine production platforms and methods, animal testing for safety evaluation is unchanged thus far. We recently developed a systems biological approach to vaccine safety evaluation where identification of specific biomarkers in a rat pre-clinical study evaluated the safety of vaccines for pandemic H5N1 influenza including Irf7, Lgals9, Lgalsbp3, Cxcl11, Timp1, Tap2, Psmb9, Psme1, Tapbp, C2, Csf1, Mx2, Zbp1, Ifrd1, Trafd1, Cxcl9, β2m, Npc1, Ngfr and Ifi47. The current study evaluated whether these 20 biomarkers could evaluate the safety, batch-to-batch and manufacturer-to-manufacturer consistency of seasonal trivalent influenza vaccine using a multiplex gene detection system. When we evaluated the influenza HA vaccine (HAv) from four different manufactures, the biomarker analysis correlated to findings from conventional animal use tests, such as abnormal toxicity test. In addition, sensitivity of toxicity detection and differences in HAvs were higher and more accurate than with conventional methods. Despite a slight decrease in body weight caused by HAv from manufacturer B that was not statistically significant, our results suggest that HAv from manufacturer B is significantly different than the other HAvs tested with regard to Lgals3bp, Tapbp, Lgals9, Irf7 and C2 gene expression in rat lungs. Using the biomarkers confirmed in this study, we predicted batch-to-batch consistency and safety of influenza vaccines within 2 days compared with the conventional safety test, which takes longer. These biomarkers will facilitate the future development of new influenza vaccines and provide an opportunity to develop in vitro methods of evaluating batch-to-batch consistency and vaccine safety as an alternative to animal testing.

Highlights

Vaccination is a beneficial and universal tool to prevent infectious disease [1]
We have demonstrated the advantage of a system biological approach using several vaccines authorized in Japan, e.g. DPT, Japanese encephalitis virus (JEV) and Influenza vaccine including H5N1 pandemic influenza vaccine [10,11,12,13]
We demonstrate that the biomarkers used to evaluate H5N1 pandemic influenza vaccine could be used to evaluate the batch-to-batch consistency and the safety of HA vaccine (HAv)

Summary

Introduction

Vaccination is a beneficial and universal tool to prevent infectious disease [1]. Because most vaccines are derived from inactivated virus, bacteria or toxoids, contamination by incomplete inactivation can cause serious adverse events. After the diphtheria toxoid (DT) immunization incident in Japan in 1950 that caused the death of 68 children and illness in over 600 infants owing to contamination by incomplete inactivation of DT [3], the abnormal toxicity test (ATT) ( known as general safety test) was introduced to the Japanese guidelines. This stated that the minimum requirement of biological products (MRBP) and all inactivated vaccines and toxoids was mandatory safety evaluation by ATT and other specific toxicity tests. Despite the increase in many vaccine production platforms, adjuvants, additives and vaccine types, safety evaluation tests in the preclinical phase and batch release have been unchanged in most countries, including in Japan

Methods

Results

Conclusion