传染性单核细胞增多症患儿外周血淋巴细胞亚群、 细胞因子、TLRs 的表达变化及其临床意义

Study on the changes and significance of lymphocytes subsets, cycokines and TLRs in children with infectious mononucleosis

摘要:

目的 检测Epstein-Barr 病毒（EBV）感染的传染性单核细胞增多症（IM）患儿外周血淋巴细胞亚群、 细胞因子及Toll 样受体7（TLR7）、TLR9 的表达变化及其临床意义。方法 选取2019 年1 月—2019 年 12 月杭州市儿童医院收治的98 例感染EBV 致IM 急性期患儿作为研究对象，另外选择50 例EBV 感染阴性 儿童作为对照组。采用流式细胞术和酶联免疫吸附试验分别检测IM 患儿急性期、恢复期（病程满1 个月后 复查）外周血淋巴细胞亚群和细胞因子水平。另外分离外周全血单个核细胞（PBMC），采用qRT-PCR 检测 TLR7 和TLR9 mRNA。结果 与对照组比较，IM 急性期患儿外周血CD3+、CD8+、白细胞介素-1β（IL- 1β）、IL-6、IL-8、IL-10、肿瘤坏死因子α（TNF-α）、γ 干扰素（IFN-γ）、TLR7 和TLR9 mRNA 相 对表达量升高（P <0.05）；CD4+、CD19+、CD16+56+ 降低（P <0.05）。IM 急性期患儿EBV DNA 载量与血清 IL-1β、TLR9 mRNA 呈正相关（r s =0.247 和0.348，P <0.05），与其他指标无关（P >0.05）。IM 患儿在恢复 期时，外周血CD3+、CD8+、IL-1β、IL-6、IL-8、IL-10、TNF-α、IFN-γ、TLR7 和TLR9 mRNA 相对 表达量降低（P <0.05）；CD4+、CD19+、CD16+56+ 升高（P <0.05）。结论 监测IM 患儿外周血淋巴细胞亚群、 细胞因子及TLR7、TLR9 的表达变化，尤其是重点关注血清IL-1β 和TLR9 mRNA 的表达，有助于了解病 情进展和转归。

Abstract:

Objective To observe the difference of peripheral blood lymphocytes subsets, cycokines and toll like receptors 7 (TLR7), TLR9 in infectious mononucleosis children infected with epstein-barr virus (EBV). Methods Ninety-eight IM children in acute phase and 50 healthy volunteers without EBV infection as control group in our hospital from January to December 2019 were enrolled into the study. The flow cytometry and ELISA methods were used to detected the percent of lymphocytes subsets and levels of IL-1β, IL-6, IL-8, IL-10, TNF-α, IFN-α and IFN-γ. Peripheral blood mononuclear cell (PBMC) were isolated from the blood, TLR7 mRNA and TLR9 mRNA in PBMC were detected by RT-qPCR. Results Compared with control group, CD3+, CD8+, IL- 1β, IL-6, IL-8, IL-10, TNF-α, IFN-γ, TLR7 mRNA and TLR9 mRNA levels in acute IM children increased, while CD4+T, CD19+, CD16+56+ declined (P < 0.05). Spearman analysis showed that the load of EBV-DNA in acute IM children would be positively related with IL-1β level and TLR9 mRNA (r = 0.247 and 0.348, P < 0.05). At the recovery stage, CD3+, CD8+, IL-1β, IL-6, IL-8, IL-10, TNF-α, IFN-γ, TLR7 mRNA and TLR9 mRNA levels declined (P < 0.05) and CD4+, CD19+, CD16+56+ increased (P < 0.05), compared with the acute IM stage. Conclusions There are some differences of peripheral blood lymphocytes subsets, cycokines and TLR7, TLR9 between acute IM stage and recovery IM stage, while especially the detection of IL-1β and TLR9 levels might be helpful to evaluate the development and clinical outcomes.