Panapasa Jack A, Cox Rebecca J, Mohn Kristin G I, Aqrawi Lara A, Brokstad Karl A
a Broegelmann Research Laboratory; Department of Clinical Sciences; University of Bergen ; Bergen , Norway.
Hum Vaccin Immunother. 2015;11(7):1663-72. doi: 10.1080/21645515.2015.1032486.
Live attenuated influenza vaccines (LAIV) can prevent influenza illness and death in children. The absence of known correlates of protection induced by LAIV requires human studies of underlying mechanisms of vaccine-induced immunity, to further elucidate the immunological processes occurring. In this study, children scheduled for elective tonsillectomy were enrolled in a clinical trial to evaluate the immune response to LAIV, in order to compare T and B cell gene expression profiles. Twenty-three children (aged 3-17 years) were divided into 4 groups; unvaccinated controls, or vaccinated intranasally with LAIV at days 3-4, 6-7, and 12-15 before tonsillectomy. Total RNA extraction was performed on tonsillar tissue and high RNA quality was assured. The samples were then analyzed using a validated RT2 Profiler PCR Array containing 84 gene-specific primers involved in B and T cell activation, proliferation, differentiation, regulation and polarization. The gene expression after LAIV vaccination was subsequently compared to the controls. We observed that at d 3-4 post vaccination, 6 genes were down-regulated, namely APC, CD3G, FASLG, IL7, CD8A and TLR1. Meanwhile at 6-7 days post vaccination, 9 genes were significantly up-regulated, including RIPK2, TGFB1, MICB, SOCS1, IL2RA, MS4A1, PTPRC, IL2 and IL8. By days 12-15 the genes RIPK2, IL4, IL12B and TLR2 were overexpressed. RIPK2 was upregulated at all 3 time points. Our data suggests an overall proliferation, differentiation and regulation of B and T cells in the tonsils following LAIV, where the majority of genes were up-regulated at days 6-7 and normalized by days 12-15. These findings may provide a first step into defining future biomarkers or correlates of protection after LAIV immunization.
减毒活流感疫苗（LAIV）可预防儿童流感疾病和死亡。由于缺乏LAIV诱导的已知保护相关因素，需要对疫苗诱导免疫的潜在机制进行人体研究，以进一步阐明所发生的免疫过程。在本研究中，计划进行择期扁桃体切除术的儿童被纳入一项临床试验，以评估对LAIV的免疫反应，以便比较T细胞和B细胞基因表达谱。23名儿童（3 - 17岁）被分为4组；未接种疫苗的对照组，或在扁桃体切除术前第3 - 4天、第6 - 7天和第12 - 15天经鼻接种LAIV。对扁桃体组织进行总RNA提取，并确保RNA质量高。然后使用经过验证的RT2 Profiler PCR Array对样本进行分析，该阵列包含84个参与B细胞和T细胞激活、增殖、分化、调节和极化的基因特异性引物。随后将LAIV接种后的基因表达与对照组进行比较。我们观察到，在接种疫苗后第3 - 4天，6个基因下调，即抗原呈递细胞（APC）、CD3G、FASLG、IL7、CD8A和TLR1。同时，在接种疫苗后第6 - 7天有9个基因显著上调，包括RIPK2、TGFB1、MICB、SOCS1、IL2RA、MS4A1、PTPRC、IL2和IL8。到第12 - 15天，RIPK2、IL4、IL12B和TLR2基因过度表达。RIPK2在所有3个时间点均上调。我们的数据表明，LAIV接种后扁桃体中B细胞和T细胞总体上发生增殖、分化和调节，其中大多数基因在第6 - 7天上调，并在第12 - 15天恢复正常。这些发现可能为定义LAIV免疫后未来的生物标志物或保护相关因素迈出了第一步。
