CAJ | 학술논문

目的研究慢性乙型肝炎病毒（HBV）感染患者外周血特异性CD8＋ T淋巴细胞（CTL）频率的变化及临床价值。方法采用乙型肝炎病毒核心抗原（HBcAg）18-27表位肽-人类白细胞抗原（HLA）-A*0201五聚体及CD8单克隆抗体，设计流式细胞技术检测 HLA-A2＋6例、HLA-A2-12例非HBV感染者、HLA-A2＋49例和HLA-A2-42例 HBV 感染者外周血中针对该肽段的特异性 CTL 数量，占总计数 CD8＋细胞数百分比表示。结果 HLA-A2-HBV感染未抗病毒治疗者特异性CTL（0．30％～14．40％，中位数1．13％，n＝20）与 HLA-A2-非HBV感染者（0．33％～3．90％，中位数1．04％，n＝12），差异无统计学意义（P＞0．05），但显著低于 HLA-A2-抗病毒治疗者（0．25％～20．30％，中位数2．11％，n＝22，P＜0．05）；HLA-A2-抗病毒治疗者特异性CTL和 HLA-A2＋未抗病毒治疗者特异性CTL（0．20％～29．90％，中位数2．22％，n＝20）均显著高于HLA-A2-非HBV感染者（P＜0．01）；HLA-A2＋抗病毒治疗者特异性CTL（0．14％～39．22％，中位数1．33％，n＝29）显著高于 HLA-A2-非 HBV感染者（P＜0．05）。HLA-A2＋未抗病毒组中血清 HBV DNA＜103 copy/mL、丙氨酸氨基转移酶小于40 U/L者特异性CTL频率增高（P＜0．05、0．01）。结论 HBV抗原肽-HLA-A*0201五聚体流式细胞技术能在体外直接检测外周血HBV特异性CTL频率的变化，其水平一定程度反映不同临床感染状态慢性 HBV感染患者 T淋巴细胞对特异性抗原表位免疫应答的差异。
목적연구만성을형간염병독（HBV）감염환자외주혈특이성CD8＋ T림파세포（CTL）빈솔적변화급림상개치。방법채용을형간염병독핵심항원（HBcAg）18-27표위태-인류백세포항원（HLA）-A*0201오취체급CD8단극륭항체，설계류식세포기술검측 HLA-A2＋6례、HLA-A2-12례비HBV감염자、HLA-A2＋49례화HLA-A2-42례 HBV 감염자외주혈중침대해태단적특이성 CTL 수량，점총계수 CD8＋세포수백분비표시。결과 HLA-A2-HBV감염미항병독치료자특이성CTL（0．30％～14．40％，중위수1．13％，n＝20）여 HLA-A2-비HBV감염자（0．33％～3．90％，중위수1．04％，n＝12），차이무통계학의의（P＞0．05），단현저저우 HLA-A2-항병독치료자（0．25％～20．30％，중위수2．11％，n＝22，P＜0．05）；HLA-A2-항병독치료자특이성CTL화 HLA-A2＋미항병독치료자특이성CTL（0．20％～29．90％，중위수2．22％，n＝20）균현저고우HLA-A2-비HBV감염자（P＜0．01）；HLA-A2＋항병독치료자특이성CTL（0．14％～39．22％，중위수1．33％，n＝29）현저고우 HLA-A2-비 HBV감염자（P＜0．05）。HLA-A2＋미항병독조중혈청 HBV DNA＜103 copy/mL、병안산안기전이매소우40 U/L자특이성CTL빈솔증고（P＜0．05、0．01）。결론 HBV항원태-HLA-A*0201오취체류식세포기술능재체외직접검측외주혈HBV특이성CTL빈솔적변화，기수평일정정도반영불동림상감염상태만성 HBV감염환자 T림파세포대특이성항원표위면역응답적차이。
Objective To investigate frequency change of peripheral blood specific CD 8+ T lymphocytes (CTL) in patients with chronic HBV infection and to evaluate its clinical value .Methods Total of 109 patients were enrolled ,including 6 cases of HLA-A2 + ,12 cases of HLA-A2 - without HBV infection ,49 cases of HLA-A2 + and 42 cases of HLA-A2 - with HBV infection .HBcAg 18-27 epitope peptide- HLA-A *0201 pentamers and CD8 mono-clonal antibody were designed to detect the number of specific CTL in peripheral blood by using flow cytometry .Re-sults The detected result of specific CTL in HLA-A2 - HBV-infected patients with no antiviral treatment (0 .30% -14 .40% ,median 1 .13% ,n=20) was not statistically different with HLA-A2 -non-HBV infected patients (0 .33% -3 .90% ,median 1 .04% ,n=12 ,P>0 .05) ,but was significantly lower than HLA-A2 - HBV-infected patients with an-tiviral treatment (0 .25% -20 .30% ,median 2 .11% ,n=22 ,P<0 .05) .The specific CTL both in HLA-A2 - HBV-in-fected patients with antiviral treatment and HLA-A2 + HBV-infected patients with no antiviral treatment (0 .20% -29 .90% ,median 2 .22% ,n=20) were significantly higher than control group (P<0 .01) .The detected result of spe-cific CTL in HLA-A2 + HBV-infected patients with antiviral treatment (0 .14% ~39 .22% ,median 1 .33% ,n= 29) was significantly higher than control group (P< 0 .05) .The number of specific CTL was increased in HLA-A2 +without antiviral therapy patients with serum HBV DNA less than 103 copy/mL and ALT less than 40 IU/L(P<0 .05 ,P< 0 .01) .Conclusion HBV antigen peptide- HLA-A *0201 pentamers could directly detect the frequency change of peripheral blood specific CTL by using flow cytometry ,and its level could reflect differences in epitope spe-cificity of the immune response of T lymphocytes in patients with different clinical status of chronic HBV infection .