HBsAg疫苗冲击树突状细胞联合乙肝免疫球蛋白对慢性乙型肝炎及HBV携带者的疗效

目的:观察HBsAg疫苗冲击的树突状细胞与乙肝免疫球蛋白联合应用对慢性乙型肝炎的疗效．方法:慢乙肝患者66例每月1次注射HBsAg疫苗(106／次)冲击的树突状细胞,乙肝免疫球蛋白200 U／次,6次为1疗程,共2疗程,治疗结束后检查肝功能,HBV DNA定量及乙肝标志．结果:HBeAg阳性慢乙肝27例治疗后有7例显示完全应答,14例显示部分应答．HBeAg阴性的慢乙肝15例中有4例出现完全应答,8例显示部分应答．慢性HBV携带者13例中5例显示完全应答,2例表现为部分应答．11例非活动性 HBsAg携带者中2例出现完全应答．结论:HBsAg疫苗冲击的树突状细胞与乙肝免疫球蛋白联合可试用于慢乙肝的治疗．     

恩替卡韦对慢性乙型肝炎患者树突状细胞功能的体外影响

目的:研究恩替卡韦(ETV)对慢性乙型肝炎(CHB)患者外周血树突状细胞(dendritic cell,DC)功能的影响.方法:体外常规分离CHB患者及健康人外周血单个核细胞,诱导扩增后常规培养.第4天将其与一定浓度的恩替卡韦共培养,第8天收获DC进行细胞表型、同种异体混合淋巴细胞反应等相关检测.结果:细胞培养8 d时DC形态分化健康对照组优于CHB ETV处理组,CHB ETV处理组优于CHB组;CHB组CD1a(35.73±3.12 vs 62.31±5.22,P<0.01),CD80(28.19±1.64 vs 45.38±3.10,P<0.01),CD83(22.24±2.14 vs 40.63±7.21,P<0.01)及HLA-DR(36.74±0.98 vs 56.05±3.89,P<0.01)表达明显低于健康对照组,而ETV处理组与CHB组相比CD83 (27.41±9.23 vs 22.24±2.14,P<0.05),CD80(32.67±7.82 vs 28.19±1.64,P<0.05)及HLA-DR(40.84±5.57 vs 36.74±0.98,P<0.01)显著高表达;淋巴细胞增殖能力测定ETV处理组DC刺激同种异体T淋巴细胞增殖能力较CHB组增强(1.53±0.09 vs 1.45±0.12,P<0.05).结论:恩替卡韦作为治疗CHB的新一代核苷类药物,除了直接抑制乙肝病毒DNA合成外,也能够增强CHB患者外周血DC的功能,通过调节机体的免疫系统发挥间接抗病毒作用.     

Entecavir for the treatment of lamivudine-refractory chronic hepatitis B patients in China

Purpose This randomized, double-blind, placebo-controlled study was undertaken to evaluate the efficacy and safety of entecavir in Chinese patients with lamivudine-refractory chronic hepatitis B. Methods One hundred forty-five lamivudine-refractory patients with chronic hepatitis B were randomized to double-blind treatment with oral entecavir 1 mg (n = 116) or placebo (n = 29) daily for 12 weeks, followed by 36 weeks of open-label entecavir treatment. The primary efficacy endpoint was the mean change from baseline in serum hepatitis B virus (HBV) DNA by polymerase chain reaction (PCR) assay at week 12. Results At week 12, the mean change from baseline in serum HBV DNA by PCR assay was –4.30 log₁₀ copies/ml for patients on entecavir compared to –0.15 log₁₀ copies/ml for patients on placebo (P < .0001). Among patients with baseline serum alanine aminotransferase (ALT) >1 × upper limit of normal (ULN), a higher proportion of entecavir than placebo patients (68% vs. 6%, respectively) achieved ALT normalization by week 12 (P < .0001). After 48 weeks of entecavir treatment, the mean change in HBV DNA by PCR assay was –5.08 log₁₀ copies/ml, and 85% of patients with baseline ALT >1 × ULN had achieved ALT normalization. The safety profile of entecavir was similar to that of placebo during the first 12 weeks of blinded dosing. Entecavir was also well tolerated during 36 weeks of open-label treatment. Conclusions Lamivudine-refractory chronic hepatitis B patients treated with entecavir demonstrated marked HBV DNA reduction and normalization of ALT in most cases. Entecavir treatment for 48 weeks was well tolerated. This clinical trial was sponsored by Bristol-Myers Squibb Company, 5 Research Parkway, Wallingford, CT, USA.     

树突状细胞治疗HBeAg阴性慢性乙型肝炎患者的临床观察

目的:观察树突状细胞疫苗对HBeAg(-)慢性乙型肝炎的治疗效果.方法:HBV抗原肽致敏的树突状细胞.1次/mo sc于17例HBeAg阴性慢性乙型肝炎患者,6次后改为HBV抗原肽和乙肝免疫球蛋白各注射1次/mo,连用6 mo.每6月复查肝功能,乙肝标志,HBV DNA定量及B超.结果:HBV DNA转阴5例(29.4%),HBV DNA下降2例(11.8%),肝功能复常4例(23.5%).结论:HBV抗原肽冲击的树突状细胞疫苗可应用于HBeAg阴性慢性乙型肝炎的治疗.     

亚硒酸钠对慢性乙型肝炎患者外周血树突状细胞的作用及机制

目的:探讨慢性肝炎患者经过亚硒酸钠作用前后树突状细胞(DC)功能的改变及机制,寻求改善DC功能的途径.方法:慢性乙肝患者28例外周血,每份血分成2份,不加亚硒酸钠处理的为A组,加亚硒酸钠者为B组.健康献血者12例为对照组(C).采用MTT法通过MLR检测DC对同种异体淋巴细胞的增殖能力及ELISA法检测细胞因子IL-12的分泌.用比色法检测3组DC内GSH-Px活性、MDA含量和膜流动性.结果:A组与C组相比,DC分泌IL-12水平(12.46±0.17 ng/L vs 21.43±0.43 ng/L,P＜0.05)和刺激自身淋巴细胞反应能力明显降低;B组比A组相比显著升高(16.93±0.32 ng/Lvs 12.46±0.17 ng/L,P＜0.05).DC细胞内GSH-Px活性A组明显低于C组(65.35±5.37 U/106细胞 vs 94.73±4.81 U/106细胞,P＜0.05),B组高于A组和C组(107.13±3.42 U/106细胞 vs 65.35±5.37,94.73±4.81 U/106细胞,P＜0.05).MDA含量A组明显高于B组和C组(1.75±0.21 U/106细胞 vs 1.09±0.17,0.82±0.13 U/106细胞,P＜0.05).B组膜流动性与A组相比保持良好.结论:体外经亚硒酸钠作用后的乙肝患者的DC可有效的刺激淋巴细胞增殖反应,并可提高IL-12分泌水平,保持膜流动性,增强对自由基损伤的抵抗能力.     

Effects of lamivudine on the function of dendritic cells derived from patients with chronic hepatitis B virus infection

AIM: To investigate if the nucleoside analogue lamivudine (LAM), a potent inhibitor of HBV replication, could restore the function of dendritic cells derived from patients with chronic hepatitis B (CHB) in an Asian population. METHODS: Dendritic cells (DCs) derived from mononuclearcytes of patients with chronic HBV infection were cultured in the presence of IL-4, granulocyte-macrophage colony-stimulating factors (GM-CSF) and gradient concentrations of LAM (0-2 mmol/L). Cell morphology was observed under light microscopy. Cell surface molecules, including HLA-DR, CD80, CD83, and CD1α, were analyzed with flow cytometry. The concentrations of IL-6 and IL-12 in the supernatant were assayed by ELISA. T cell proliferation was assayed by methyl thiazolyl tetrazolium (MTT). RESULTS: The expression of CD1α on DC treated with 0.5 mmol/L LAM (LAM-DC 0.5 mmol/L) was significantly higher than that of DC untreated with LAM (54.1 ± 4.21 vs 33.57 ± 3.14, P < 0.05), and so was the expression of CD83 (20.24 ± 2.51 vs 12.83 ± 2.12, P < 0.05) as well as the expression of HLA-DR (74.5 ± 5.16 vs 52.8 ± 2.51, P < 0.05). Compared with control group, LAM-DC group (0.5 mmol/L) secreted significantly more IL-12 (910 ± 91.5 vs 268 ± 34.3 pg/mL, P < 0.05), had lower levels of IL-6 in the culture supernatant (28 ± 2.6 vs 55 ± 7.36 pg/mL, P < 0.05), markedly enhanced the stimulatory capacity in the allogeneic mixed leukocyte reaction (MLR) (1.87 ± 0.6 vs 1.24 ± 0.51, P < 0.05).CONCLUSION: The lower expression of phenotypic molecules and impaired allogeneic mixed lymphocyte reaction function of dendritic cells derived from patients with HBV infection could be restored in vitro by incubation with LAM.     

慢性乙型肝炎患者在HBV不同抗原负载下树突状细胞功能的变化

目的观察在不同HBV抗原负载下树突状细胞(DC)功能的变化及在核苷(酸)类药物抗病毒治疗后DC的功能变化。方法分离17例慢性乙型肝炎患者外周血单个核细胞(PBMC),并在不同抗原负载下(HBsAg、HBcAg、HBsAg联合HBcAg)诱导DC分化成熟,用流式细胞技术测定DC表面共刺激分子CD80、CD83、CD1a及HLA-Ⅱ类分子HLA-DR表达水平,同时用淋巴细胞增殖试验评估DC功能。结果与负载HBsAg诱导相比,HBcAg、HBsAg联合HBcAg诱导下DC共刺激分子表达明显增强(HLA-DR:85.12±1.55比98.37±1.27比99.21±1.33;CD1a:15.69±2.46比16.25±2.33比42.20±1.10;CD80:71.88±6.38比80.74±3.23比94.70±2.77;CD83:32.64±2.77比42.55±2.88比44.16±1.89)。淋巴细胞增殖试验提示DC功能改善更显著,尤其HBsAg联合HBcAg诱导DC功能改善更加明显(HBV DNA阳性:7.29±0.17比7.99±0.43比8.56±0.31;HBV DNA阴性:7.48±0.30比8.22±0.41比8.78±0.31)。将17例慢性乙型肝炎患者在核苷(酸)类药物抗病毒治疗前后DC功能进行比较,经抗病毒治疗在充分抑制HBV DNA复制的情况下,DC功能的恢复更趋于完善(HLA-DR:99.21±1.33比99.82±2.67;CD1a:42.20±1.10比71.33±5.89;CD80:94.70±2.77比96.42±3.56;CD83:44.16±1.89比68.34±2.11)。结论 HBsAg联合HBcAg诱导DC细胞功能明显得到恢复和增强,HBV DNA阴性时诱导自体DC功能恢复更趋完善。     

CpG-ODN联合HBsAg对慢性乙型肝炎患者外周血树突状细胞表型和功能的影响

目的　研究含非甲基化CpG基序的免疫刺激寡核苷酸 (CpG- ODN)与重组HBsAg对慢性乙型肝炎患者 (CHB)外周血树突状细胞 (dendriticcell ,DC)表型和功能的影响。方法　以重组人GM CSF、IL 4自CHB患者和健康者外周血单个核细胞诱导扩增DC ;以CpG ODN和HBsAg单独或联合刺激DC ,并与TNF α比较 ,评价其对DC表达表面分子HLA DR、CD86、CD1a ,分泌IL 12p70以及刺激同种T细胞增殖能力的影响 ;同时检测血浆TGF- β、IFN- γ含量。 结果　与PBS组比 ,CpG- ODN单用或联合HBsAg均能明显提高CHB患者DC表面分子HLA DR的表达 ,使IL- 12分泌增加 ,刺激同种T细胞增殖的能力亦增强 ,CpG ODN联合HBsAg尚能明显提高CD1a的表达 ;CpG- ODN的上述刺激作用类似于TNF α ;CHB患者血浆TGF- β、IFN -γ含量明显高于正常对照。 结论　CpG -ODN与TNF α一样能够促进CHB患者外周血DC分化和成熟 ;CpG- ODN与HBsAg联合刺激能协同增强DC的特异性抗原递呈作用 ;CHB患者的细胞因子环境可能是DC功能沉默的重要原因。     

复合干扰素α治疗对慢性乙型肝炎患者浆样树突状细胞及淋巴细胞亚群的影响

目的动态观察慢性乙型肝炎患者重组复合干扰素(CIFN)α治疗过程中PDC、淋巴细胞亚群的变化。方法采用流式细胞分析技术检测23例慢性乙型肝炎患者CIFN治疗过程中外周血浆样树突状细胞(PDC)和淋巴细胞亚群百分比及数量。结果经CIFN治疗后,患者的应答率为43.5%。外周血PDC的百分比和绝对数显著减少(P值均<0.05),有效组与无效组PDC百分比和绝对数平均值在同一时间点无明显差别。同时CD3(?)细胞、CD4(?) T淋巴细胞、B淋巴细胞明显减少(P值均<0.05),CD4/CD8明显下降(P<0.05),但在治疗的24周及随访24周后患者CD8(?)T淋巴细胞、自然杀伤细胞有增加(P<0.05)。结论经CIFN治疗,慢性乙型肝炎患者外周血PDC、CD8(?)T淋巴细胞、自然杀伤细胞数量出现变化,部分患者可以获得持续性病毒学及生化学应答。     

拉米夫定体外对慢性乙型肝炎患者树突状细胞功能的影响

目的:体外研究不同浓度拉米夫定(lamivu- dine,LAM)对慢性乙型肝炎(chronic hepatitis B,CHB)患者树突状细胞(dendritic cell,DC)功能影响．方法:分离慢乙肝患者外周血单核细胞,在含GM-CSF IL-4及不同浓度LAM(0,0．125, 0．25,0．5,1,2 mmol／L)培养条件下制备DC．观察DC形态学变化并用MTT法测定DC刺激同种异体淋巴细胞增殖能力,流式细胞仪(FCM) 测定其细胞表型分子CD1a,CD83,CD80及 HLA-DR的表达,ELISA法检测培养上清中 IL-12和IL-6含量．结果:在LAM 0．5 mmol／L组,DC表型分子 CD83,CD1a,HLA-DR的表达最高,CD80与 LAM未处理组相比无明显差异．与LAM未处理组相比,LAM 0．5 mmol／L处理组DC膜表面分子CD1a,CD83,HLA-DR表达增高(CD1a: 54．0±9．2 vs 33．6±10．1,P<0．05;CD83:20．3 ±6．1 vs 11．8±6．2,P<0．05;HLA-DR:74．5± 7．1 vs 52．9±7．7,P<0．05);其上清液中IL-12 分泌水平增高(810．0±91．5 ng／L vs 268．0± 34．3 ng／L,P<0．05),IL-6则显著降低(28．1± 2．6 ng／L vs 55．3±7．4 ng／L,P<0．05);刺激同种异体淋巴细胞增殖能力(SI)增强(1．9±0．6 vs 1．2±0．5,P<0．05)．结论:LAM体外可增强慢乙肝患者树突状细胞活性．     

恩替卡韦抗病毒治疗4周HBV DNA载量可预测48周病毒学应答

目的评价恩替卡韦抗病毒的疗效并分析其预测因素。方法 61例慢性乙型肝炎患者接受恩替卡韦抗病毒治疗48周,分别于基线、4、12、24和48周检测患者肝功能、HBV血清标志物、HBV DNA定量。结果治疗48周时ALT复常率和HBV DNA转阴率分别达95.1%和91.8%,HBsAg定量较基线显著下降（P〈0.05）;48周时完全病毒学应答率与第4周时HBV DNA载量有显著相关性。结论恩替卡韦抗病毒安全、有效;第4周时HBV DNA载量对48周时完全病毒学应答率有很好的预测作用。     

恩替卡韦联合苦参素治疗HBeAg阳性慢性乙型肝炎的疗效观察

[目的]观察恩替卡韦(ETV)联合苦参素治疗HBeAg阳性慢性乙型肝炎(CHB)的疗效。[方法]92例HBeAg阳性CHB患者随机分为2组,治疗组48例,每天服用ETV 0.5 mg,苦参素片0.6 g;对照组44例,每天服用ETV 0.5 mg。分别观察服药12、24、48周时的治疗效果。[结果]对照组和治疗组在12、24、48周的HBV DNA阴转率分别为29.55%、40.91%、63.64%和43.75%、58.33%、83.33%,2组比较在12、24周时差异无统计学意义(P0.05),而在48周时差异有统计学意义(P0.05)。对照组和治疗组的HBeAg转阴率,在12、24周时分别为36.37%、43.75%和50.00%、64.59%,2组比较P0.05,而在48周时分别为59.09%和81.25%(P0.05)。对照组和治疗组的HBeAg/HBeAb血清转换率,12、24周时分别为29.55%、37.50%和40.91%、52.09%,2组比较P0.05,而在48周时分别为47.73%和68.75%,差异有统计学意义(P0.05)。[结论]ETV联合苦参素治疗CHB可明显提高抗病毒治疗效果。     

慢性乙型肝炎树突状细胞表型和功能的变化与免疫耐受

目的 观察慢性乙型肝炎病毒（HBV）感染者树突状细胞（DC）形态和功能的改变。方法 从13例慢性乙型肝炎患者和11例健康人外周血中分离和培养DC,观察DC的形态,用流式细胞仪检测DC表面标记HLA－DR、CD1a、CD80和CD86的表达,用^3H-TdR掺入法测定DC诱导混合性淋巴细胞反应（MLR）的能力。结果 正常DC较慢性乙型肝炎患者在形态上更为典型,不规则,HLA－DR、CD80和CD86分子的表达水平较高（P＜0.05）,诱导MLR的能力较强（P＜0.05）。结论 慢性乙型肝炎患者外周血DC处于不完全成熟状态,其免疫刺激能力较低。     

恩替卡韦治疗藏族 HBeAg 阳性的慢性乙型肝炎患者临床疗效分析

目的：评价恩替卡韦治疗藏族 HBeAg 阳性的慢性乙型肝炎患者的临床疗效。方法60例 HBeAg 阳性的藏族慢性乙型肝炎患者接受口服恩替卡韦0.5 mg/d 治疗48 w。每3个月随访1次,治疗12周及48周时复查肝功能及 HBV DNA,并记录不良反应事件。结果在观察期结束时,有80%患者达到 HBV DNA 阴转,有75.4%患者达到 ALT 复常,有7%患者达到 HBeAg血清学转阴,有3%患者达到 HBeAg 血清学转换。在治疗过程中没有患者出现 HBsAg 转阴。结论恩替卡韦治疗藏族 HBeAg 阳性慢乙型肝炎患者具有持续抑制病毒复制的作用,安全性较好。     

慢性乙型肝炎患者外周血树突状细胞表型及抗原提呈能力与HBV载量的关系

目的探讨慢性乙型肝炎(CHB)患者外周血树突状细胞(DCs)表型及抗原提呈能力与HBV载量的关系.方法采集23例CHB患者和8例健康人的抗凝外周静脉血,分离外周血单个核细胞(PBMCs),在重组人白细胞介素4和重组人粒细胞-巨噬细胞集落刺激因子的作用下培养使DCs增殖、成熟,以间接免疫荧光流式细胞技术分别检测DCs表面CD80、CD86、HLA-DR及ICAM-1的表达;将培养成熟的DCs与HBsAg共同孵育,用丝裂霉素C处理后再与自体PBMCs共同培养,在培养结束前12 h加入^3H-TDR,收集细胞,以β液闪计数仪测定cpm值;同期用定量聚合酶链反应技术测定CHB患者外周血HBV载量.结果患者DCs表面CD86、HLA-DR和ICAM-1的表达水平及DCs的抗原提呈能力均显著低于健康对照组;CD80、CD86、HLA-DR及ICAM-1的表达与HBV载量呈显著负相关(分别为P＜0.01,P＜0.01,P＜0.001和P＜0.001);DCs的抗原提呈能力也与HBV载量呈显著负相关(为P＜0.001).结论CHB患者外周血DCs的成熟和功能存在障碍,DCs的抗原提呈能力与血液中HBV的载量密切相关,并可能对HBV的清除产生重要的影响.     

恩替卡韦治疗e抗原阴性慢性乙型肝炎停药后复发的分析

目的了解HBeAg阴性的慢性乙型肝炎(CHB)患者使用恩替卡韦抗病毒治疗的疗程及停药的标准。方法回顾性分析2006年1月至2015年12月复旦大学附属中山医院肝炎门诊确诊为HBeAg阴性的CHB并予恩替卡韦抗病毒治疗后停药6月以上的患者121例,观察其停药6个月、12个月的HBV DNA水平、肝功能。结果停药6个月后有33%的患者HBV DNA5×10~2拷贝/mL,19.8%患者出现ALT升高。停药12个月后累计有45.5%的患者HBV DNA5×10~2拷贝/mL,30.6%患者出现ALT升高,用药大于5年的患者复发率最低。此外,基线HBV DNA水平2×10~5拷贝/mL的患者停药后6个月、12个月的复发率较基线水平10~7拷贝/mL者低。结论 HBeAg阴性的CHB患者使用恩替卡韦治疗停药复发的比率随着抗病毒疗程的延长而减少,且基线HBV DNA水平与复发率相关。     

恩替卡韦联合苦参素治疗慢性乙型肝炎48周疗效观察

目的：观察恩替卡韦联合苦参素治疗慢性乙型肝炎患者48周的临床疗效。方法选择慢性乙型肝炎患者100例,随机分为两组。治疗组52例应用恩替卡韦片联合苦参素片治疗;对照组48例仅服用恩替卡韦片,观察48周。结果在治疗48周时,联合组患者血清 ALT 复常率、HBeAg 转阴率和 HBV DNA 转阴率分别为80.77%、82.69%和82.67%,均优于对照组的72.92%、58.33%和62.50%,差异均具有统计学意义（P〈0.05）。结论恩替卡韦联合苦参素治疗慢性乙型肝炎具有协同作用,且无不良反应。     

替比夫定与恩替卡韦治疗HBeAg阳性慢性乙型肝炎4年随访观察

目的比较替比夫定(LdT)和恩替卡韦(ETV)治疗HBeAg阳性的CHB,哪一个能较早地实现有限疗程。方法 60例HBeAg阳性的CHB患者随机分为LdT组和ETV组。在服药后第12周、24周及以后每24周复查,直至治疗和随访结束(4年,192周),检测血清HBV DNA定量、HBV血清标志物、肝功能、血清肌酸激酶。观察两组192周治疗结束时应答率及持续应答率以及患者达到停药标准所需的时间及费用。结果 192周时LdT组和ETV组治疗结束时应答率及持续应答率为43.3%(13/30)vs 16.7%(5/30)、36.7%(11/30)vs 10.0%(3/30),差异有统计学意义(P<0.05)。达到停药标准LdT组平均治疗时间为167.2周,费用28 089.6元,而ETV组平均治疗时间为186.4周,费用50 887.2元。结论治疗HBeAg阳性的CHB患者,实现有限疗程LdT更具有优势。     

Long-term treatment outcomes of clevudine in antiviral-naive patients with chronic hepatitis B

AIM: To evaluate the treatment outcomes of clevudine compared with entecavir in antiviral-naive patients with chronic hepatitis B (CHB).METHODS: We retrospectively analyzed the clinical data of CHB patients treated with clevudine 30 mg/d and compared their clinical outcomes with patients treated with entecavir 0.5 mg/d. The biochemical response, as assessed by serum alanine aminotransferase (ALT) activity, virologic response, as assessed by serum hepatitis B virus DNA (HBV DNA) titer, serologic response, as assessed by hepatitis B e antigen (HBeAg) status, and virologic breakthrough with genotypic mutations were assessed.RESULTS: Two-hundred and fifty-four patients [clevudine (n = 118) vs entecavir (n = 136)] were enrolled. In clevudine-treated patients, the cumulative rates of serum ALT normalization were 83.9% at week 48 and 91.5% at week 96 (80.9% and 91.2% in the entecavir group, respectively), the mean titer changes in serum HBV DNA were -6.03 and -6.55 log₁₀ copies/mL (-6.35 and -6.86 log₁₀ copies/mL, respectively, in the entecavir group), and the cumulative non-detection rates of serum HBV DNA were 72.6% and 83.1% (74.4% and 83.8%, respectively, in the entecavir group). These results were similar to those of entecavir-treated patients. The cumulative rates of HBeAg seroconversion were 21.8% at week 48 and 25.0% at week 96 in patients treated with clevudine, which was similar to patients treated with entecavir (22.8% and 27.7%, respectively). The virologic breakthrough in the clevudine group occurred in 9 (7.6%) patients at weeks 48 and 15 (12.7%) patients at week 96, which primarily corresponded to genotypic mutations of rtM204I and/or rtL180M. There was no virologic breakthrough in the entecavir group.CONCLUSION: In antiviral-naive CHB patients, long-term treatment outcomes of clevudine were not inferior to those of entecavir, except for virologic breakthrough.