摘要
目的研究EB病毒（Epstein—Barr virus，EBV）慢性活动性感染（CAEBV）、急性感染（AEBV）以及正常儿童的外周血单个核细胞（PBMC）的EBV—DNA水平，以及EBV感染细胞类型的差异，探讨其与CAEBV临床表型的关系。方法收集2004年3月至2008年4月在我院住院的CAEBV患儿10例，AEBV患儿13例，以及正常儿童12例的外周血单个核细胞，应用实时荧光定量PCR法检测EBV—DNA水平，并对EBV—DNA阳性的CAEBV和AEBV及正常儿童，用免疫磁珠法分选各种淋巴细胞后进行EBV编码的RNA-1（EBV encoding RNA-1，EBER-1）探针荧光原位杂交（FISH）确定EBV感染细胞的类型。结果CAEBV组EBV—DNA载量为[（6．8×10^7）±（1．1×10^8）]／ml，AEBV组EBV-DNA载量为[（1．3×10^6）±（1．6×10^6）]／ml，两组比较差异有统计学意义，CAEBV组PBMC的EBV—DNA水平明显高于AEBV组（P〈0．01）；7例CAEBV患儿做细胞分选及FISH后，发现EBV不仅可以引起B细胞感染，而且还引起NK细胞、CD4^＋和CD8^＋T细胞不同程度的感染，临床表现为反复或持续的传染性单核细胞增多症（IM）样症状。6例患儿以感染T细胞为主，其中1例以CD8’T细胞感染为主，临床表现除高热，肝脾淋巴结大，伴严重的血液系统一系或三系降低外，还并发了爆发性的致死性T淋巴细胞增殖综合征而死亡。1例以NK细胞感染为主，临床表现还伴有对蚊虫叮咬高度敏感且IgE高达2500U／ml。AEBV组7例患儿均显示感染B淋巴细胞，临床表现为可以痊愈的IM。6例正常儿童均为阴性。结论CAEBV患儿体内存在更多的EBV复制和不同的EBV感染细胞类型，实时荧光定量PCR检测EBV-DNA水平并测定EBV感染的淋巴细胞类型有可能协助CAEBV临床个体化诊治和评估病情进展。
Objective To study the difference in the EBV-DNA level in peripheral blood mononuclear cells （PBMC） and the type of Epstein-Barr virus （EBV）-infected cells in pediatric patients with chronic active EBV （CAEBV） infection, acute EBV infection （AEBV） and healthy children, and to analyze the relationship between the above difference and the clinical manifestation of CAEBV. Method Real-time fluorescent quantitative polymerase chain reaction （PCR） was used to detect the EBV-DNA levels in peripheral blood mononuelear cells （ PBMC ） in 12 normal children, 10 pediatric patients with CAEBV infection and 13 pediatric patients with AEBV infection in our hospital between March 2004 and April 2008. Immunomagnetic bead cell fraetionation and fluorescent in situ hybridization （FISH） by EBV encoding RNA-1 （EBER-1） probe were used in the healthy children, EBV-DNA positive CAEBV patients and AEBV patients to detect the type of EBV-infeeted cells. Result The average EBV-DNA level in CAEBV patients＇PBMC was （6.8 ×10^7 ＋ 1.1 ×l0^8） copies/ml, while the average EBV-DNA level of AEBV patients＇ PBMC was （ 1.3 ×10^6 ＋ 1.6× 10^6 ） copies/ml. The average EBV-DNA level of CAEBV infected patients＇ PBMC was significantly higher than that of AEBV infected patients＇ PBMC （P 〈 0. 01 ）. The cell fractionation and FISH in seven CAEBV patients showed that EBV in CAEBV patients infected not only B cells, but NK cells and CI4 ^＋ and CD8 ＋ T cells to different degree, and these patients presented recurrent and persistent infectious mononucleosis （IM） -like symptoms. In 6 CAEBV patients infection mainly occurred to T cells, in one case, infection occurred mainly in CD8 ^＋ T cells, and the patient died from fulminant and deadly T lymphocytes proliferative syndrome except presenting firstly high fever, enlargment of the liver, spleen, lymphnode and the severe decrease of one or three kinds of blood cells. In 1 CAEBV patient the infection was mainly found in NK cells, who presented wit
出处
《中华儿科杂志》
CAS
CSCD
北大核心
2011年第7期495-500,共6页
Chinese Journal of Pediatrics
基金
中国医学科学院北京协和医院临床重点科研项目
关键词
疱疹病毒4型
人
免疫磁化分离
实时荧光定量PCR
荧光原位杂交
Herpesvirus 4, human
Immunomagnetic separation
Real-time fluorescent quantitative PCR
Fluorescent in situ hybridization
