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目的观察EB病毒(EBV)原发感染后裂解期和潜伏期病毒抗原肽特异性T细胞功能变化,分析外周血EBV载量(VL)与特异性T细胞反应间的关系。方法采集首都医科大学附属北京儿童医院2006年6月至2006年7月收住院的6例EBV相关性传染性单核细胞增多症(EBV-associated IM)患儿和4例保健门诊EBV血清学抗体转化阳性的健康对照组患儿的外周血。利用磁珠分离法(MACS)检测HLA型别。在4个时间点,利用酶联免疫斑点检测法(ELISPOT)及荧光定量PCR法检测特异性T细胞分泌γ-干扰素(INF-γ)水平和外周血EBV VL。结果观察组6例特异性T细胞分泌INF-γ水平高于对照组(P<0.05)。观察组病例病程2个月时裂解期肽BMLF1特异性T细胞分泌INF-γ水平较感染急性期时下降,而潜伏期肽LMP2的结果呈上升趋势,在病程10个月时两者均较前下降,裂解肽特异性T细胞反应的下降幅度大于潜伏期肽特异性T细胞,且其水平较感染急性期和病程2个月时差异有统计学意义(P<0.05);病程20个月与病程10个月结果差异无统计学意义(P>0.05)。观察组4例(4/6)患儿外周血EBV VL为阴性。4个时间点均可检测到HLA-A11限制性BMLF1特异性T细胞分泌INF-γ阳性细胞。结论 EBV原发感染后裂解期与潜伏期肽特异性T细胞反应的变化不同,外周血EBV VL与特异性T细胞反应可能无关。HLA限制性BMLF1特异性T细胞可能在EBV原发感染后起保护作用。
Objective To observe the functional changes of peptide specific T lymphocytes (PBMCs) during the lysis and incubation of Epstein-Barr virus (EBV) primary infection, and to analyze the relationship between the EBV load (VL) in peripheral blood and specific T cell responses. Methods Six children with EBV-associated infectious mononucleosis (EBV-associated IM) admitted to Beijing Children’s Hospital Affiliated to Capital Medical University from June 2006 to July 2006 were enrolled in this study. Four EBV serological antibodies Peripheral blood from children with positive conversion to healthy controls. HLA typing was detected by magnetic bead separation (MACS). At 4 time points, the level of IFN-γ secreted by specific T cells and EBV VL in peripheral blood were detected by ELISPOT and fluorescence quantitative PCR. Results The level of INF-γ secreted by 6 T cells in the observation group was higher than that in the control group (P <0.05). In the observation group, the level of INF-γ secreted by peptide BMLF1-specific T cells in lysis group was lower than that in acute phase at 2 months, and the LMP2 latency was on the rise. Both of them were lower at 10 months , The decrease of cleavage peptide specific T-cell response was greater than that of latent peptide-specific T cells, and the difference was statistically significant (P <0.05) compared with the acute phase and the 2-month duration of infection; There was no significant difference in monthly results (P> 0.05). The EBV VL in peripheral blood of 4 cases (4/6) in the observation group was negative. At 4 time points, HLA-A11-restricted BMLF1-specific T cells secreted INF-γ-positive cells. Conclusion The changes of EBV-specific T cell responses during the lysis of EBV primary infection are different from those of EBV-specific T cell responses. HLA-restricted BMLF1-specific T cells may have protective effects following primary EBV infection.