劉坤 楊亞萍 段銀環(huán) 趙俊梅 孫謝文

·論著·

慢性HBV感染者外周血T細(xì)胞亞群與病毒載量、HBeAg的相關(guān)性分析

劉坤 楊亞萍 段銀環(huán) 趙俊梅 孫謝文

目的分析慢性HBV感染者外周血T細(xì)胞亞群與病毒載量、HBeAg的相關(guān)性。方法以40例慢性乙型肝炎(CHB)、35例慢性重型乙型肝炎(CSHB)、30例肝硬化(LC)及32例正常對照者為研究對象，采用流式細(xì)胞儀和熒光定量PCR法分別檢測各組外周血T細(xì)胞亞群、HBV DNA載量。結(jié)果CHB組僅CD+4亞群明顯降低，與對照組比較，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；CSHB及LC組CD+3、CD+4亞群、CD+4/CD+8比值呈逐漸降低趨勢，CD+8亞群呈逐漸增高趨勢，與對照組比較差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。HBV DNA載量與CD+4亞群、CD+4/CD+8比值呈負(fù)相關(guān)(r=-0.638，-0.778，P<0.05)，與CD+8亞群呈正相關(guān)(r=0.647，P<0.05)，與CD+3無關(guān)。與HBeAg陽性患者比較，HBeAg陰性患者外周血CD+3+、CD+4和CD+8亞群顯著降低(P<0.05)。結(jié)論慢性HBV感染者隨著疾病加重細(xì)胞免疫功能進(jìn)行性降低，與病毒復(fù)制水平密切相關(guān)，這是導(dǎo)致疾病慢性化的原因之一。外周血T細(xì)胞亞群變化趨勢可為判斷疾病轉(zhuǎn)歸及預(yù)后提供可靠指標(biāo)。

乙型肝炎病毒；慢性肝炎；T細(xì)胞亞群

1 資料與方法

1.1 一般資料 選取2011年6月至2013年10月期間我院收治的HBsAg陽性患者105例，其中CHB患者40例、CSHB患者35例、LC患者30例。105例中，男78例，女27例;年齡15～76歲，平均年齡(43.5±5.7)歲；均符合2010年《慢性乙型肝炎防治指南》對慢性乙型肝炎及肝硬化的診斷標(biāo)準(zhǔn)[3]。所有患者6個(gè)月內(nèi)均未接受抗病毒治療。排除其他病毒引起的肝炎、HIV陽性者及肝癌、酒精性肝病等。同時(shí)選取我院健康體檢者32例為對照組，男20例，女12例；年齡21～68歲，平均年齡(39.8±4.3)歲；均經(jīng)查體證實(shí)HBsAg陰性，肝功能正常。

1.2 檢測方法

1.2.2 HBV DNA載量測定：使用美國ABI7300型基因擴(kuò)增儀，采用實(shí)時(shí)熒光定量PCR法檢測標(biāo)本，試劑盒由上海生工生物工程技術(shù)有限公司提供。HBV DNA≤103copies/ml為陰性，HBV DNA>103～105copies/ml為低載量，HBV DNA 105～107copies/ml為中載量，HBV DNA>107copies/ml為高載量。

2 結(jié)果

組別CD+3CD+4CD+8CD+4/CD+8對照組(n=32)74.35±4.5642.28±4.3624.65±3.241.72±0.39CHB組(n=40)72.12±4.4737.66±4.21*25.06±3.451.50±0.36CSHB組(n=35)66.35±4.47*#30.21±3.89*#32.68±3.99*#0.92±0.32*#LC組(n=30)61.22±4.13*△20.63±3.65*△37.22±5.13*△0.55±0.21*△

注：與對照組比較，*P<0.05；與CHB組比較，#P<0.05；與CSHB組比較，△P<0.05

表2不同HBV DNA載量與HBV患者T細(xì)胞亞群比較

組別CD+3CD+4CD+8CD+4/CD+8對照組(n=32)74.35±4.5642.28±4.3624.65±3.241.72±0.39陰性組(n=25)73.56±4.3240.76±4.4424.98±3.591.63±0.34低載量組(n=20)67.56±4.55*33.24±3.32*28.12±3.87*1.18±0.26*中載量組(n=30)62.22±4.13*#27.15±3.95*#32.21±5.04*#0.84±0.21*#高載量組(n=30)57.27±4.13*△17.56±4.13*△37.21±5.04*△0.47±0.17*△

注：與對照組和陰性組比較，*P<0.05；與低載量組比較，#P<0.05；與中載量組比較，△P<0.05

表3 HBeAg陽性HBV感染者與HBeAg陰性HBV感染者T細(xì)胞亞群的比較

組別CD+3CD+4CD+8CD+4/CD+8對照組(n=32)74.35±4.5642.28±4.3624.65±3.241.72±0.39HBeAg陽性組(n=45)72.37±4.2440.87±4.4525.13±3.351.63±0.42HBeAg陰性組(n=60)60.23±3.76*24.15±3.34*34.12±4.22*0.71±0.34*

注：與HBeAg陽性組比較，*P<0.05

3 討論

總之，如果我們能夠在慢性乙肝逐步進(jìn)展至慢重肝及肝硬化的早期提高機(jī)體的細(xì)胞免疫功能就有可能成功阻斷HBV的復(fù)制，這也為慢性乙肝的治療、疾病預(yù)測及改善預(yù)后提供了理論依據(jù)。

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ThecorrelationbetweenTlymphocytesubsetsinperipheralbloodandHBVDNAlevelsaswellasHBeAginpatientswithchronichepatitisBvirusinfection

LIUKun,YANGYaping,DUANYinhuan,etal.DepartmentofLiverDiseases,TheThirdHospitalofQinhuangdaoCity,Hebei,Qinhuangdao066000,China

ObjectiveTo investigate the relationship between T-lymphocyte subsets and HBV DNA levels,HBeAg in patients with chronic hepatitis B virus (HBV) infection.MethodsThe 40 cases of chronic hepatitis(CHB),35 cases of chronic severe hepatitis B (CSHB),30 cases of liver cirrhosis (LC) and 32 healthy subjects were enrolled in the study.The T lymphocyte subsets and HBV DNA load in peripheral blood were detected by flow cytometry and fluorescence quantitative PCR for all the patients and healthy subjects.ResultsThe CD+4subset was significantly decreased in CHB group,as compared with that in control group (P<0.05);the CD+3,CD+4subsets,the ratio of CD+4/CD+8in CSHB group and LC group were gradually decreased,however,CD+8subset was gradually increased,as compared with those in control group (P<0.05).HBV DNA load was negatively correlated to CD+4subset,ratio of CD+4/CD+8(r=-0.638,-0.778,P<0.05),however,which was positively correlated to CD+8subset (r=0.647,P<0.05),moreover,which was not related with CD+3subset.As compared with those of patients with positive HBeAg,the CD+3 subset,CD+4subset and CD+8subset in peripheral blood of patients with negative HBeAg were significantly decreased (P<0.05).ConclusionThe cellular immune function in patients with chronic HBV infection is progressively decreased with the aggravation of disease condition,which is closely correlated to the levels of virus replication,and it is one of the reasons that results in chronicity of disease.The changing trend of T lymphocyte subsets in peripheral blood can be regarded as a reliable index for evaluating disease’s prognosis.

hepatitis B virus;chronic hepatitis;T-lymphocytes subsets

10.3969/j.issn.1002-7386.2014.13.006

066000 河北省秦皇島市第三醫(yī)院肝病科

R 512.62

A

1002-7386(2014)13-1940-03

2014-02-09)