儿童非肿瘤性EB病毒感染相关疾病的血清学和免疫学特征分析

目的 了解非肿瘤性EB病毒(Epstein-Barr virus,EBV)感染患儿不同临床类型的EBV抗体、EBV-DNA水平及免疫功能的变化特点,为EBV感染防治提供依据.方法 回顾性分析第四军医大学西京医院住院的资料完整的103例EBV感染患儿的临床资料,按临床疾病类型分为3组:传染性单核细胞增多症(infectious mononualeosis,IM)组68例;慢性活动性EBV感染(chronic active Epstein-Barr virus infection,CAEBV)组13例;EBV相关噬血细胞性淋巴组织细胞增多症(Epstein-Barr virus-related hemophagocytic lymphohistiocytosis,EBV-HLH)组22例.检测并比较各组的EBV抗体、EBV-DNA、免疫球蛋白水平、淋巴细胞亚群及补体系列的变化.选择同期正常儿童26例为对照组,检测其免疫球蛋白水平、淋巴细胞亚群及补体系列,并与其余3组比较.结果 (1)CAEBV组、EBV-HLH组的C3、C4水平明显低于其余2组(P均<0.05);EBV-HLH组、CAEBV组、IM 组及对照组的IgA、IgG、IgE水平均逐渐升高,EBV-HLH组、CAEBV组均低于对照组,差异均有统计学意义(P均<0.05),IM组略低于对照组,但差异无统计学意义(P>0.05).(2)IM组CD8+T细胞较其余3组显著升高,差异均有统计学意义(P均<0.05);EBV-HLH组的总T细胞、CD4+、CD8+、CD4+/ CD8+、NK及B细胞比其余3组均显著下降(P均<0.05).(3)IM组及CAEBV组各项EBV抗体阳性率均高于EBV-HLH(P均<0.05);EBV-HLH组的EBV-DNA水平均高于IM组及CAEBV组,差异均有统计学意义(P均<0.05).结论 血清中的EBV-DNA水平与疾病种类及严重程度呈正相关;EBV感染导致的IM、CAEBV 及EBV-HLH 3种疾病发病机制均与免疫功能紊乱有关;动态监测EBV载量和细胞免疫功能可反映疾病状态和进展的风险.     

EB病毒相关噬血细胞性淋巴组织细胞增生症患儿的EB病毒感染特征

目的 了解儿童EB病毒(EBV)相关噬血细胞性淋巴组织细胞增生症(EBV-HLH)患儿的EBV血清学抗体及病毒复制水平等特征.方法 对67例EBV-HLH患儿和60例原发性EBV感染所致的传染性单核细胞增多症(EBV-IM)患儿的临床资料进行分析,EBV特异性抗体和血清EBV DNA载量分别采用间接免疫荧光法和荧光定量PCR测定.结果 EBV-HLH患儿EBV特异性抗体结果:EBV-CA-IgM、EBV-CA-IgG、EBV-EA-IgG和EBV-NA-IgG的阳性率分别为28.8％、100.0％、51.5％和78.8％,EBV-VCA-IgG高亲和力为78.9％,低亲和力为12.1％;血清学抗体结果显示,71.2％的患儿为EBV再激活感染,其余为急性原发EBV感染;45.5％的EBV-HLH患儿可在血清中检测到EBV DNA,其拷贝数中位数为1.976×103 copies·L-1.EBV-IM患儿的EBV-CA-IgM、EBV-CA-IgG、EBV-EA-IgG和EBV-NA-IgG阳性率分别为100.0％、100.0％、58.3％和26.7％,EBV-VCA-IgG高亲和力为18.3％,低亲和力为81.7％,EBV DNA阳性率为10.0％,DNA拷贝数均值为8.495 copies·1-1.IM患儿均为EBV原发感染状态.结论 EBV-HLH可发生在EBV原发感染或既往EBV感染再激活时期,但多数患儿由既往EBV感染再激活所致.EBV-HLH患儿血清中EBV复制水平显著高于EBV-IM患儿.     

Prevalence of Epstein-Barr Viral DNA among children at a single hospital in Suzhou,China

ObjectiveThis study aimed to describe the prevalence of Epstein-Barr virus (EBV)-DNA among children in Suzhou, and to explore the association between plasma EBV load and disease diagnosis.MethodsAll children admitted to the Children's Hospital of Soochow University between January 2018 and September 2020 and subjected to the plasma EBV-DNA assay were included. The authors retrospectively collected demographic and discharge diagnostic information of the participants, and ascribed the disease distribution characteristics of children with positive plasma EBV-DNA by age and viral load.ResultsA total of 38,175 patients underwent plasma EBV-DNA PCR assay, of which 2786 (7.3%) had EBV-DNA in their plasma. Children aged 3–4 years had a high prevalence of EBV infection. Plasma EBV positivity was common with infectious mononucleosis (IM, 40.0%), respiratory infection (20.1%), atypical EBV infection (14.2%), acute leukemia (6.4%), hemophagocytic lymphohistiocytosis (HLH, 4.8%), and idiopathic thrombocytopenic purpura (ITP, 2.9%). With increasing age, plasma EBV positivity was more common in children with IM and atypical EBV infection. However, an inverse correlation was observed in children with respiratory infections and ITP. High levels of EBV loads were more likely to occur in HLH, IM, and atypical EBV infection, especially in HLH. However, lower viral loads were found in respiratory infection and acute leukemia.ConclusionsThis is a large sample study that revealed the prevalence of plasma EBV-DNA levels in children of various ages and presenting illnesses.     

慢性活动性EB病毒感染患儿外周血单个核细胞EB病毒DNA及感染细胞类型检测的临床意义

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[(6.8×107)±(1.1 x 108)]/ml,AEBV组EBV-DNA载量为[(1.3×106)±(1.6×106)]/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2500 U/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.

Abstract：

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 mononuclear 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 fractionation 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-infected cells.Result The average EBV-DNA level in CAEBV patients'PBMC was(6.8×107±1.1×108)copies/ml, while the average EBV-DNA level of AEBV patients' PBMC was(1.3×106±1.6×106)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 CD4+ 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 with hypersensitivity to mosquito biting and high IgE level (2500 U/ml).But EBV in seven AEBV patients infection was found only in B cells who presented with only IM for one time and no EBV-infected PBMC were found in the remaining 6 healthy children. Conclusion There are much more EBV replications and different EBV-infected cell types in CAEBV patients. Detection of EBV-DNA level by real-time fluorescent quantitative PCR and the detection of the type of EBV-infected cells may help in diagnosis, treatment and development evaluation of children with CAEBV infection.     

儿童EB病毒相关噬血淋巴组织细胞增生症26例

目的探讨儿童EB病毒相关噬血淋巴组织细胞增生综合症(EBV-HLH)的病原学特征及临床实验室特点、治疗及预后相关因素。方法收集2003年8月~2006年8月我院收治的26例EBV-HLH患儿临床及实验室资料,追踪并回顾性分析。结果本组15例患儿EBV-HLH由重症传染性单核细胞增多症(IM)快速进展所致;8例为既往感染的再激活,3例由慢性活动性EB病毒感染(CAEBV)发展而来。实验室检查:26例患儿有不同程度的外周血三系减低及凝血功能异常并伴有脂质代谢紊乱,骨髓中出现吞噬血细胞现象。治疗及转归:在26例患儿中,16例患儿死亡,总死亡率61.5%。化疗组19例死亡总死亡率47.4%,非化疗组7例死亡率100%,2组有显著性差异(P=0.023)。脏器损害大于4个部位与死亡相关(P=0.009)。在10例存活患儿中,至少9例仍存在EB病毒的活动。结论EBV-HLH病情凶险,预后差,以多脏器损害者更差。早期诊断并尽早开始化疗可以提高患儿的存活率。缓解后可继续处于CAEBV感染状态下,甚至可转化成EBV相关淋巴瘤。     

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??Abstract??Objectives??To study the function of the EBV lytic and latent peptides specific CTLs and analyze the correlations between peripheral EBV Viral Load ??VL?? and the CTLs responses from primary to persistent infection. Methods ??Six patients with infectious mononucleosis ??IM?? were enrolled in the study along with four healthy children with past EBV infection as controls. Human leukocyte antigen ??HLA?? identification was performed by magnetic beads cell sorting ??MACS??. Enzyme-linked immunospot ??ELISPOT?? and Real-time PCR were performed. To determine INF-γ lever secreted by specific T-lymphoayte & the peripheral EBV VL. Results??ELISPOT and EBV VL in peripheral blood were performed in acute phase?? 2 months?? 10 months and 20 months after admission for all the cases except one case who lost of follow-up in 20 months. The CTLs responses against lytic peptides and latent peptides in patients were significantly increased ??P < 0.05?? compared with control??. Among patients with IM the CTLs responses against lytic peptides decayed in contrast against latent peptides increased first then decayed from primary to persistent infection. All of them decreased sharply in 10 months after admission ??P < 0.05?? compared with 2 months??.Then they were at a low level or undetectable ultimately ??P > 0.05?? compare 20 months with 10 months??. The results of EBV VL in peripheral blood of case No. 4 were 3.52×103?? 6.75×102?? 2×102 and negative in the acute phase?? 2 months?? 10 months and 20 months respectively?? and the results of case No. 6 were 2×103 in the acute phase and negative in other time. Four cases in the rest had negative results of EBV VL in peripheral blood all the time. The CTLs response against HLA restricted lytic peptide BMLF1 was positive after primary to persistent infection. Conclusions??There are differences between the CTLs responses against the lytic peptides and the latent peptides and no correlations between EBV VL in peripheral blood and the CTLs responses. The abundant result of the distribution of CTLs response against BMLF1 may play a protective role after primary infection.     

Primary Epstein-Barr virus infection in 2-year-old children: report of 3 cases

Some children less than four years old have Epstein-Barr virus (EBV)-induced infectious mononucleosis (IM). Because primary EBV infection in infants and young children is usually asymptomatic or subclinical, EBV infection diagnosis may not be easy among young children. To illustrate the clinical characteristics and diagnostic procedures for EBV infection in young children, the authors report herein three cases of primary EBV infection in two-year-old children with an evaluation of their initial clinical symptoms. The results showed that the common initial clinical manifestations are puffy eyelids and hepatosplenomegaly, and that these signs suggest a tentative diagnosis of IM. In conclusion, EBV capsid immunoglobulin (Ig)M antibodies and atypical lymphocytes are useful diagnostic measurements in very young children with symptoms suggestive of IM.     

儿童EB病毒相关性噬血细胞淋巴组织细胞增生症的临床特征和预后危险因素分析

目的 分析儿童EB病毒相关性噬血细胞淋巴组织细胞增生症(EBV-HLH)的临床特征,探讨影响其预后的危险因素.方法 采用回顾性调查方法 ,对2003年5月至2008年9月收治的62例EBV-HLH患儿临床特征、血清学、病毒载量、病理改变、基因筛查及预后资料进行系统分析.根据随访的生存情况分为生存组和死亡组,采用单因素和多因素Logistic回归分析影响预后的危险因素.结果 (1) 62例患儿中,男36例,女26例,发病年龄2个月～14岁,26例(41.9%)在婴幼儿期发病,38例(61.3%)发病是由于EBV感染再激活所致;(2) 所有患儿均有持续或间断发热,至少有外周血两系减低.伴有肝肿大52例,脾肿大45例,淋巴结病43例.58例患儿血清白蛋白降低,52例血清铁蛋白增高,大部分患儿伴有凝血功能异常和脂质代谢紊乱.48例诊断时骨髓中出现吞噬血细胞现象;31例EBV-HLH患儿中14例血清中EBV-DNA检测阳性,病毒载量拷贝数在5.12×10~2～7.69×10~7/ml之间(平均10~(3.9)/ml);(3) 在3例EBV-HLH的PRF1基因外显子编码区发现3个杂合错义突变,这3个突变均导致氨基酸改变(C102F,S108N和T450M),1例患儿为复合杂合错义突变(S108N和T450M),从遗传学上可明确诊断为家族性噬血细胞淋巴组织细胞增生症亚型2(FHL2);(4) 随访57例病例中35例(61.4%)死亡,其中21例经过HLH-94或04治疗方案.15例是在住院后2个月内死亡.死亡病例相比存活病例白蛋白降低和部分凝血激酶时间延长(P均<0.05).多因素Logistic回归分析显示病程大于1个月、未进行免疫化疗、白蛋白≤25 g/L和深部出血与EBV-HLH的预后呈显著相关性(P均<0.05).结论 EBV-HLH患儿病情严重,预后凶险,病死率高;多数病例由于EBV感染再激活所致;早期诊断并尽早开始化疗可以提高患儿的存活率;病程大于1个月、未进行化疗、白蛋白降低和深部出血是影响其预后的主要危险因素.     

761例住院儿童EB病毒感染分析

目的：了解儿童EB病毒（EBV）感染情况,并分析其相关疾病谱,从而为EBV感染及相关疾病的防治提供科学的理论依据。方法：采用real-time PCR法检测2010年8月至2011年7月收治的761例（年龄22 d至14岁）疑似EBV感染儿童血浆中EBV-DNA载量,并对EBV-DNA检查结果及相关疾病进行统计学分析。结果：761例血浆标本中EBV-DNA阳性标本109例,阳性率为14.3%;不同年龄组EBV-DNA阳性检出率差异有统计学意义（P<0.05）,其中婴儿组（<1岁）的阳性检出率最低（P<0.05）;不同季节间阳性检出率差异有统计学意义（P<0.05）,其中夏季阳性检出率高于冬季（P<0.05）。109例阳性标本的EBV-DNA载量范围为2.13～6.69,中位数为3.72。对62例EBV-DNA阳性住院患儿最终临床诊断分析得出,呼吸系统疾病占39%,主要为急性支气管炎、急性上呼吸道感染及急性支气管肺炎。结论：不同年龄组及不同季节间EBV-DNA阳性检出率不同;儿童EBV感染相关疾病以呼吸系统疾病为主;Real-time PCR法检测血浆EBV-DNA有助于临床上EBV感染的早期诊断。     

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目的探讨儿童非典型EB病毒(EBV)感染的临床特点。方法2000年3月至2005年4月,深圳市儿童医院在住院患儿中检测出EBVVCAIgM和(或)EBVDNA阳性共690例,其中传染性单核细胞增多症422例,另外268例为非典型EBV感染,对其临床特点进行回顾性分析。结果儿童非典型EBV感染以呼吸道感染最为多见(191/268,71.3%),其它为皮炎13例(4.9%)、腹泻病10例(3.7%)、血小板减少性紫癜10例(3.7%)、肾炎或肾病7例(2.6%)、肠系膜或颈淋巴结炎8例(3.0%)、结膜炎7例(2.6%),川崎病、嗜血细胞综合征、中枢神经系统感染各4例(1.5%),心肌炎、播散性脑脊髓炎各2例(0.7%),再生障碍性贫血、坏死性淋巴结炎、腮腺炎、高IgM血症、嗜酸细胞增多症各1例(0.4%),诊断不明1例(0.4%)。其中嗜血细胞综合征患儿有2例死亡。结论儿童非典型EBV感染症状多样,累及系统多,预后不一。EBVVCAIgM或EBVDNA检测有利于早期诊断,并进行合理治疗。