中华肝脏病杂志
中華肝髒病雜誌
중화간장병잡지
CHINESE JOURNAL OF HEPATOLOGY
2008年
10期
739-742
,共4页
韩亚萍%李军%万玉峰%孔练花%蔡洁%董莉%刘源%陈念%黄祖瑚
韓亞萍%李軍%萬玉峰%孔練花%蔡潔%董莉%劉源%陳唸%黃祖瑚
한아평%리군%만옥봉%공련화%채길%동리%류원%진념%황조호
肝炎e抗原,乙型%单核细胞%Toll样受体2%固有免疫
肝炎e抗原,乙型%單覈細胞%Toll樣受體2%固有免疫
간염e항원,을형%단핵세포%Toll양수체2%고유면역
Hepatitis B e antigens%Menocytes%Toll-like receptor 2%Innate immunity
目的 探讨HBeAg对外周血单核细胞表面Toll样受体2(TLR2)表达的影响.方法 对68例慢性乙型肝炎(CHB)患者(其中HBeAg与HBV DNA刚性37例,HBeAg阴性、HBVDNA阳性14例,HBeAg与HBV DNA阴性17例)和16名健康人的外周血肝素抗凝,加入Pam3CSK4在37℃、5%CO2条件下刺激3 h,用特异性TLR2单克隆抗体标记,经流式细胞仪检测CD14+细胞表面TLR2表达的百分率;比较刺激前后各组之间的差异.定量检测HBV DNA和血清HBV标志物,分析TLR2表达水平与HBeAg、HBV DNA的关系.用HBeAg与健康人及HBeAg阴性CHB患者的抗凝血在37℃、5%CO2条件下共培养6 h,用流式细胞仪定量分析HBeAg刺激后CD14+细胞表面TLR2的表达水平. 结果 HBeAg阳性CHB患者外周血CD14+细胞TLR2的表达率为47.57%±21.40%,明显低于健康对照组(76.51%±7.46%)和HBeAg阴性组(HBV DNA阳性组为73.2%±14.2%、HBV DNA阴性组为75.2%±11.3%,P<0.05);TLR2的表达水平在HBeAg阴性的HBV DNA阳性或阴性CHB患者组与健康对照组的差异无统计学意义.HBeAg阳性CHB患者外周血单核细胞经Pam3CSK4刺激后,TLR2的表达率明显升高,大部分患者TLR2的表达水平能够达到健康人或HBeAg阴性患者未经配体刺激的水平.健康人或HBeAg阴性CHB患者的外周血与HBeAg共孵育6 h后,CD14+细胞表面TLR2表达水平较HBeAg刺激前明显下降(P<0.05).结论 HBeAg阳性的CHB患者外周血CD14+细胞TLR2的表达水平明显低于HBeAg阴性患者和健康人,HBeAg能够抑制CD14+细胞TLR2的表达,提示HBeAg能够引起CHB患者外周血单核细胞表面TLR2表达的下调,这可能是HBV持续感染的重要因素之一;CHB患者HBVDNA水平可能不影响单核细胞表面TLR2的表达;Pam3CSK4可以明显提高HBeAg阳性CHB患者单核细胞表面TLR2的表达水平,TLR2配体有可能成为CHB免疫治疗的一个潜在靶位.
目的 探討HBeAg對外週血單覈細胞錶麵Toll樣受體2(TLR2)錶達的影響.方法 對68例慢性乙型肝炎(CHB)患者(其中HBeAg與HBV DNA剛性37例,HBeAg陰性、HBVDNA暘性14例,HBeAg與HBV DNA陰性17例)和16名健康人的外週血肝素抗凝,加入Pam3CSK4在37℃、5%CO2條件下刺激3 h,用特異性TLR2單剋隆抗體標記,經流式細胞儀檢測CD14+細胞錶麵TLR2錶達的百分率;比較刺激前後各組之間的差異.定量檢測HBV DNA和血清HBV標誌物,分析TLR2錶達水平與HBeAg、HBV DNA的關繫.用HBeAg與健康人及HBeAg陰性CHB患者的抗凝血在37℃、5%CO2條件下共培養6 h,用流式細胞儀定量分析HBeAg刺激後CD14+細胞錶麵TLR2的錶達水平. 結果 HBeAg暘性CHB患者外週血CD14+細胞TLR2的錶達率為47.57%±21.40%,明顯低于健康對照組(76.51%±7.46%)和HBeAg陰性組(HBV DNA暘性組為73.2%±14.2%、HBV DNA陰性組為75.2%±11.3%,P<0.05);TLR2的錶達水平在HBeAg陰性的HBV DNA暘性或陰性CHB患者組與健康對照組的差異無統計學意義.HBeAg暘性CHB患者外週血單覈細胞經Pam3CSK4刺激後,TLR2的錶達率明顯升高,大部分患者TLR2的錶達水平能夠達到健康人或HBeAg陰性患者未經配體刺激的水平.健康人或HBeAg陰性CHB患者的外週血與HBeAg共孵育6 h後,CD14+細胞錶麵TLR2錶達水平較HBeAg刺激前明顯下降(P<0.05).結論 HBeAg暘性的CHB患者外週血CD14+細胞TLR2的錶達水平明顯低于HBeAg陰性患者和健康人,HBeAg能夠抑製CD14+細胞TLR2的錶達,提示HBeAg能夠引起CHB患者外週血單覈細胞錶麵TLR2錶達的下調,這可能是HBV持續感染的重要因素之一;CHB患者HBVDNA水平可能不影響單覈細胞錶麵TLR2的錶達;Pam3CSK4可以明顯提高HBeAg暘性CHB患者單覈細胞錶麵TLR2的錶達水平,TLR2配體有可能成為CHB免疫治療的一箇潛在靶位.
목적 탐토HBeAg대외주혈단핵세포표면Toll양수체2(TLR2)표체적영향.방법 대68례만성을형간염(CHB)환자(기중HBeAg여HBV DNA강성37례,HBeAg음성、HBVDNA양성14례,HBeAg여HBV DNA음성17례)화16명건강인적외주혈간소항응,가입Pam3CSK4재37℃、5%CO2조건하자격3 h,용특이성TLR2단극륭항체표기,경류식세포의검측CD14+세포표면TLR2표체적백분솔;비교자격전후각조지간적차이.정량검측HBV DNA화혈청HBV표지물,분석TLR2표체수평여HBeAg、HBV DNA적관계.용HBeAg여건강인급HBeAg음성CHB환자적항응혈재37℃、5%CO2조건하공배양6 h,용류식세포의정량분석HBeAg자격후CD14+세포표면TLR2적표체수평. 결과 HBeAg양성CHB환자외주혈CD14+세포TLR2적표체솔위47.57%±21.40%,명현저우건강대조조(76.51%±7.46%)화HBeAg음성조(HBV DNA양성조위73.2%±14.2%、HBV DNA음성조위75.2%±11.3%,P<0.05);TLR2적표체수평재HBeAg음성적HBV DNA양성혹음성CHB환자조여건강대조조적차이무통계학의의.HBeAg양성CHB환자외주혈단핵세포경Pam3CSK4자격후,TLR2적표체솔명현승고,대부분환자TLR2적표체수평능구체도건강인혹HBeAg음성환자미경배체자격적수평.건강인혹HBeAg음성CHB환자적외주혈여HBeAg공부육6 h후,CD14+세포표면TLR2표체수평교HBeAg자격전명현하강(P<0.05).결론 HBeAg양성적CHB환자외주혈CD14+세포TLR2적표체수평명현저우HBeAg음성환자화건강인,HBeAg능구억제CD14+세포TLR2적표체,제시HBeAg능구인기CHB환자외주혈단핵세포표면TLR2표체적하조,저가능시HBV지속감염적중요인소지일;CHB환자HBVDNA수평가능불영향단핵세포표면TLR2적표체;Pam3CSK4가이명현제고HBeAg양성CHB환자단핵세포표면TLR2적표체수평,TLR2배체유가능성위CHB면역치료적일개잠재파위.
Objective In order to investigate the relationship among the toll-like receptor 2 (TLR2),hepatitis B e antigen and HBV DNA, the expression levels of TLR2 on peripheral blood monocytes of chronic hepatitis B (CHB) patients as well as on their monocytes stimulated by ligand of TLR2 (Pam3CSK4) and HBeAg were analyzed. Methods Sixty-eight adults with CHB were enrolled, including 37 HBeAg-positive patients, 17 HBeAg-negative and HBV DNA negative patients, and 14 HBeAg-negative and HBV DNA positive patients. Sixteen healthy volunteers were also studied as controls. TLR2 expression levels on their peripheral blood monocytes stimulated with Pam3CSK4 or not stimulated were analyzed by FACS Calibur. The relationship of the expression levels of TLR2, HBeAg and HBV DNA were also analyzed. The level of TLR2 on peripheral blood monocytes of healthy volunteers and HBeAg-negative CHB patients stimulated by HBeAg was examined for six hours. Results The TLR2 expression levels on CD14+ cells were significantly reduced in HBeAg-positive patients (47.57% ± 21.40 %) compared to both healthy volunteers (76.51% ±7.46%) and HBeAg-negative patients (HBV DNA positive group 73.2% ± 14.2%, HBV DNA negative group 75.2% ± 11.3%); but there was no difference between those of the HBeAg-negative patients and the healthy volunteers. Expression levels of TLR2 on monocytes stimulated by TLR2 ligand in HBeAg-positive patients were obviously increased, and reached the basic levels of the healthy volunteers and the HBeAg-negative patients. The expression levels of TLR2 on monocytes stimulated by HBeAg of the healthy volunteers and the HBeAg-negative patients were markedly reduced. Conclusions In the presence of HBeAg, HBV down-regulates the expressions of TLR2 on CD14+ cells from peripheral blood, and there is no correlation between HBV-DNA and TLR2. Pam3CSK4 can boost the TLR2 expression in HBeAg-positive patients. The pro-posed interaction between HBV and TLR2 may provide an important clue to explain the reasons of the establishment of persistent HBV infection.
