劉坤 楊亞萍 刁青 陳璀 孫謝文

·論著·

慢性重型乙型肝炎患者外周血DNT細(xì)胞及T細(xì)胞亞群的表達(dá)及意義

劉坤 楊亞萍 刁青 陳璀 孫謝文

目的研究外周血CD+3CD-4CD-8(DNT)細(xì)胞及T細(xì)胞亞群在慢性重型乙型肝炎患者的表達(dá)及意義。方法正常對(duì)照者32例、慢性乙型肝炎(CHB)40例、慢性重型乙型肝炎(CSHB)35例患者為研究對(duì)象，采用流式細(xì)胞儀檢測(cè)各組外周血DNT細(xì)胞及T細(xì)胞亞群。結(jié)果CHB組和CSHB組DNT細(xì)胞占T淋巴細(xì)胞的百分比與對(duì)照組比較明顯增高(P<0.05)；而CSHB組增高程度更為顯著，與CHB組比較差異亦有統(tǒng)計(jì)學(xué)意義(P<0.05)。HBV DNA載量與DNT細(xì)胞占T淋巴細(xì)胞的百分比呈正相關(guān)(r=0.742，P<0.05)。CHB組與對(duì)照組比較，僅CD+4亞群明顯降低，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；CSHB組和CHB組比較，CD+4亞群及CD+4/CD+8明顯降低，CD+8亞群明顯升高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。結(jié)論慢性重型乙型肝炎患者DNT細(xì)胞顯著增高及T細(xì)胞亞群比例異常可能是慢性乙型肝炎發(fā)生發(fā)展的重要因素。

慢性重型乙型肝炎；DNT細(xì)胞；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月期間我院收治的慢性HBV感染者75例，其中，男55例，女20例；年齡16～73歲，中位年齡40.8歲；CHB患者40例和CSHB患者35例。均符合2010年《慢性乙型肝炎防治指南》對(duì)慢性乙型肝炎的診斷標(biāo)準(zhǔn)[5]。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(yáng)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(duì)照組，其中男20例，女12例；年齡21～68歲，中位年齡39.8歲；均經(jīng)查體證實(shí)HBsAg陰性，肝功能正常。

1.2 檢測(cè)方法

1.2.2 HBV DNA載量測(cè)定：使用美國(guó)ABI7300型基因擴(kuò)增儀，采用實(shí)時(shí)熒光定量PCR法檢測(cè)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

2.1 3組DNT細(xì)胞分布比較 CHB組和CSHB組DNT細(xì)胞絕對(duì)數(shù)與對(duì)照組比較差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)。DNT細(xì)胞占T淋巴細(xì)胞的百分比與對(duì)照組比較明顯增高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)；而CSHB組增高程度更為顯著，與CHB組比較差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。見(jiàn)表1。

表1各組DNT細(xì)胞分布比較

組別DNT細(xì)胞絕對(duì)數(shù)(個(gè)/mm3)DNT(%)1對(duì)照組(n=32)118±596±4 CHB組(n=40)112±558±4*CSHB組(n=35)124±6211±5*#

注：與對(duì)照組比較，*P<0.05；與CHB組比較，#P<0.05

2.2 不同HBV DNA載量與HBV患者DNT細(xì)胞分布比較 陰性組與對(duì)照組比較，DNT細(xì)胞占T淋巴細(xì)胞的百分比差異無(wú)統(tǒng)計(jì)學(xué)意義(P>0.05)；隨著HBV DNA載量增高，DNT細(xì)胞百分比逐漸升高，差異有統(tǒng)計(jì)學(xué)意義(P<0.05)。見(jiàn)表2。

表2不同HBV DNA載量與HBV患者DNT細(xì)胞分布比較

組別DNT對(duì)照組(n=32)6±4陰性組(n=8)6±4低載量組(n=12) 8±4*#中載量組(n=32) 10±5*#△高載量組(n=23) 11±5*#☆

注：與對(duì)照組比較，*P<0.05；與陰性組比較，#P<0.05；與低載量組比較，△P<0.05；與中載量組比較，☆P<0.05

2.3 HBV DNA載量與HBV患者DNT細(xì)胞分布的相關(guān)性 HBV DNA載量的對(duì)數(shù)值與DNT細(xì)胞占T淋巴細(xì)胞的百分比呈正相關(guān)(r=0.742，P<0.05)。

組別CD+4CD+8CD+4/CD+8對(duì)照組(n=32)42±4 25±31.7±0.4CHB組(n=40)38±4*25±31.5±0.4CSHB組(n=35)30±4*# 33±4*#0.9±0.3*#

注：與對(duì)照組比較，*P<0.05；與CHB組比較，#P<0.05

3 討論

慢性乙型肝炎患者由于病毒持續(xù)復(fù)制可誘導(dǎo)機(jī)體產(chǎn)生各種細(xì)胞因子、炎性介質(zhì)及宿主免疫病理反應(yīng)，這些復(fù)雜作用最終造成大塊肝組織壞死進(jìn)而發(fā)生為慢性重型乙型肝炎、肝衰竭甚至多臟器功能衰竭，患者病死率高達(dá)40%～60%。研究表明，淋巴細(xì)胞介導(dǎo)的細(xì)胞免疫功能紊亂是影響慢性乙型肝炎病變發(fā)展及預(yù)后的重要因素，其中免疫調(diào)節(jié)細(xì)胞在這一病理?yè)p傷過(guò)程中發(fā)揮重要作用[6]。

總之，慢性重型乙型肝炎患者存在DNT細(xì)胞高表達(dá)及T淋巴細(xì)胞比例失衡，這是導(dǎo)致機(jī)體免疫功能低下，感染慢性化的主要原因。

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ExpressionandsignificanceofdoublenegativeTcellsandlymphocytesubtypeinpatientswithchronicseverehepatitisB

LIUKun,YANGYaping,DIAOQing,etal.DepartmentofLiverDiseases,TheThirdHospitalofQinhuangdaoCity,Hebei,Qinhuangdao066001,China

ObjectiveTo investigate the expression and significance of CD+3CD-4CD-8(DNT)cells and lymphocyte subtype in patients with chronic severe hepatitis B.MethodsThe serum samples from 32 healthy subjects,40 patients with chronic hepatitis(CHB)and 35 patients with chronic severe hepatitis B (CSHB) were collected in this study.The DNT cells and lymphocyte subtype in peripheral blood were detected by flow cytometry.ResultsThe percent of DNT cells in CHB and CSHB groups [(8.44±4.25)% and (10.78±5.33)%,respectively] were significantly higher than those in control group [(6.23±3.77)%,P<0.05],moreover,the increase extent in CSHB group was more obvious than that in CHB group (P<0.05).The HBV DNA load was positively correlated to DNT cells (r=0.742,P<0.05).The CD+4+ T lymphocytes in CHB group were significantly decreased,as compared with those in control group (P<0.05).The percentages of CD+4T lymphocytes and the ratio of CD+4/CD+8in CSHB group were significantly decreased,however,CD+8T lymphocytes were significantly increased,as compared with those in CHB group (P<0.05).ConclusionThe obvious increase of DNT cells and the abnormality of T lymphocyte subset percentage in patients with chronic severe hepatitis B may be one of important reasons for the pathogenesis and development of chronic hepatitis B．

chronic severe hepatitis B;double negative T cells;T-lymphocytes subsets

10.3969/j.issn.1002-7386.2014.15.002

066001 河北省秦皇島市第三醫(yī)院肝病科

R 512.62

A

1002-7386(2014)15-2248-03

2014-01-07)