慢性HBV感染者PBMC分泌IFN-α功能初探及慢乙肝患者长期核苷（酸）药物抗病毒疗效
发布时间：2018-07-26 17:55
【摘要】：背景 乙型肝炎病毒(hepatitis B virus, HBV)感染是一种严重影响公众健康的传染病。我国HBV感染者有9300多万人,其中3000多万为慢性乙型肝炎(chronic hepatitis B, CHB)患者。目前HBV慢性感染的免疫机制尚未明确,但越来越多的研究显示,天然免疫在其中起到重要作用。其中,树突状细胞(dendritic cell, DC)和单核细胞(monocyte, MO)作为重要的天然免疫细胞,其在HBV感染慢性化机制中发挥的作用值得探究。干扰素a (interferon-alpha, IFN-a)是人体抵御病毒感染的重要屏障,CHB患者体内是否存在IFN-a分泌下降具有很重要的临床意义。人体95%以上的IFN-a由浆细胞样树突状细胞(plasmacytoid dendric cell, pDC)分泌。单核细胞(monocyte)亦是分泌IFN-a的重要来源,单核细胞是否存在IFN-a的分泌下降未见报道。 CHB患者最主要的治疗方法为抗病毒治疗。核苷(酸)类似物(nucleos(t)ide analogue, NUCs)药物的问世掀开了抗病毒治疗新的篇章。国内外对CHB患者抗病毒治疗做了大量研究,但多来源于Ⅲ期临床试验结果,国内尚需进一步丰富拉米夫定(1amivudine, LAM)、替比夫定(telbivudine, LdT)及恩替卡韦(entecavir, ETV)在临床真实世界中抗病毒治疗效果的长期随访资料。研究目的 本研究第一部分旨在检测慢性HBV感染者外周血单个核细胞(peripheral blood mononuclear cell, PBMC)分泌IFN-a的情况,以探究其外周血pDC和单核细胞功能是否存在缺陷。 本研究第二部分旨在探究真实世界中CHB患者长期口服NUCs药物抗病毒治疗的疗效,并比较LAM, LdT,及ETV的疗效差异。 方法 本研究第一部分前瞻性纳入2012年7月至2013年3月北京协和医院肝炎门诊就诊的慢性乙型肝炎患者16例,与之年龄相匹配的HBV携带者16例。健康对照组来源于北京协和医院健康体检中心,与之年龄相匹配的健康人18例。采集血样,分离PBMC,分别与CPG寡聚脱氧核苷酸2216(cytosine guanine dinucleotides oligodeoxy nucleotides, CPG ODN2216)、聚肌苷酸胞苷酸(Polyinosinic-Polycytidylicacid,poly I：C)刺激共培养,并用酶联免疫吸附法(enzyme linked immunosorbent assay,Elisa)的方法测量其分泌IFN-a量。 本研究第二部分回顾性分析2008年6月至2013年4月于北京协和医院肝炎门诊就诊的CHB患者87例,最长随访时间为192周,每3个月评估其丙氨酸氨基转氨酶(alanine aminotransferase, ALT)水平、乙型肝炎病毒脱氧核糖核酸(hepatitisB virus Deoxyribonucleicacid, HBV DNA)和乙型肝炎病毒表面标志物的变化情况,探究LAM,LdT,及ETV单药治疗的病毒学应答,生化学应答,e抗原转换及耐药情况。 结果 第一部分 经ODN2216刺激后,CHB组患者PBMC分泌IFN-α量平均为31.20(7.33～44.04)pg/ml, HBV携带者组PBMC分泌IFN-α量平均为109.91(13.74～240.27)pg/ml,健康对照组PBMC分泌IFN-α量平均为107.95(48.59～227.33)。CHB患者组明显低于健康对照组,具有统计学差异(P=0.007)。CHB患者组明显低于HBV携带者组,具有统计学差异(P=0.027) 经poly(I：C)刺激后,CHB患者组PBMC分泌IFN-α量平均为37.50(4.40～44.45)pg/ml,HBV携带者组PBMC分泌IFN-a量平均为10.44(3.46～36.08)pg/ml,健康对照组PBMC分泌IFN-a量平均为7.06(2.89～22.80)pg/ml,三组之间无明显差异(P=0.427)。 第二部分 1.LAM组患者治疗24周、48周、96周和144周HBV DNA抑制率为68.8%,63.2%,80.0%,81.8%。ALT复常率为72.2%,80.0%,90.9%,83.3%。e抗原血清学转换率分别为11.1%,18.2%,33.3%,37.5%。 2.LdT组患者治疗24周、48周、96周和144周HBV DNA抑制率为90.9%,90.9%,66.7%,80.0%。ALT复常率为63.6%,88.9%,100.0%,80.0%。e抗原血清学转换率分别为12.5%,14.3%,40.0%,50.0%。 3.ETV组患者治疗24周、48周、96周和144周HBV DNA抑制率为84.8%,83.3%,100%,85.7%,ALT复常率为80.6%,88.9%,70.0%,75.0%。e抗原血清学转换率分别为0%,10.0%,22.2%,28.6%。 4.病毒学耐药方面,LAM累积病毒学突破率为40.9%,LdT组累积病毒学突破率为15.4%,ETV组累积病毒学突破率为5.0%。 结论 第一部分： 1.HBV携带者和健康对照PBMC中树突状细胞分泌IFN-α的量显著高于单核细胞。 2.HBV携带者与健康对照相比,PBMC中树突细胞分泌IFN-α无显著差异。 3.CHB患者相比HBV携带者、健康对照,PBMC中树突状细胞分泌IFN-α明显下降。 4.CHB患者、HBV携带者相比健康对照,PBMC中单核细胞分泌IFN-α无显著差异。 第二部分： 1.病毒学应答方面,随访至144周,三组药物治疗无统计学差异。ETV组获得较高的HBV DNA抑制率,其次为LAM组和LdT组。 2.生化学应答方面,随访至144周,三组药物治疗无统计学差异。LAM组获得了较高的ALT复常率,其次为LdT组和ETV组。 3.e抗原血清学转换方面,随访至144周,三组药物治疗无统计学差异。LdT组获得了较高的e抗原血清学转换率,其次为ETV组和LAM组。 4.病毒学耐药方面,LAM累积病毒学突破率远远高于其余两组,LdT次之,ETV组最低。
[Abstract]:background
Hepatitis B virus (hepatitis B virus, HBV) infection is an infectious disease that seriously affects public health. There are about 93000000 people with HBV infection in China, of which about 30000000 are chronic hepatitis B (chronic hepatitis B, CHB). The immune mechanism of HBV chronic infection is not clear, but more and more studies show that the natural immune system is in it. It plays an important role. Among them, dendritic cell (DC) and mononuclear cells (monocyte, MO) are important natural immune cells, and their role in the chronicity mechanism of HBV infection is worth exploring. Interferon a (interferon-alpha, IFN-a) is an important barrier to the human body against virus infection, and IFN-a in CHB patients is in the presence of IFN-a. Secretory decline is of great clinical significance. More than 95% of IFN-a is secreted by plasma cell like dendritic cells (plasmacytoid dendric cell, pDC). Monocyte (monocyte) is also an important source of secretion of IFN-a, and the presence of IFN-a in mononuclear cells has not been reported.
The most important treatment for CHB patients is antiviral therapy. The advent of nucleoside (acid) (acid) analogues (nucleos (T) ide analogue, NUCs) has opened a new chapter in antiviral therapy. A large number of studies have been made on antiviral therapy for CHB patients at home and abroad, but many of them are derived from phase III clinical test results, and further enrichment of lamivudine (1amivudine) is needed at home. (LAM), telbivudine (LdT) and Cave (entecavir, ETV) in the clinical real world antiviral treatment of long-term follow-up data.
The first part of this study was designed to detect the secretion of IFN-a in peripheral blood mononuclear cells (peripheral blood mononuclear cell, PBMC) in patients with chronic HBV infection, in order to explore whether the function of pDC and mononuclear cells in peripheral blood has defects.
The second part of this study aims to explore the efficacy of long term oral NUCs antiviral therapy in CHB patients in the real world, and compare the difference in efficacy between LAM, LdT and ETV.
Method
The first part of the study prospectively included 16 chronic hepatitis B patients in the Peking Union Medical College Hospital hepatitis clinic from July 2012 to March 2013. 16 cases of HBV carriers were matched with age. The healthy control group was derived from the healthy physical examination center of the hospital and 18 healthy people matched with the age. The blood samples were collected and the PBMC was separated. Not with CPG oligo deoxydeoxynucleotides 2216 (cytosine guanine dinucleotides Oligodeoxy nucleotides, CPG ODN2216), and polyoside acid cytidine acid (Polyinosinic-Polycytidylicacid, poly I:C) stimulated co culture, and the method of enzyme linked immunosorbent assay was used to measure its secretion.
The second part of this study analyzed 87 cases of CHB patients in Peking Union Medical College Hospital hepatitis clinic from June 2008 to April 2013. The longest follow-up time was 192 weeks, and the level of alanine aminotransferase (alanine aminotransferase, ALT) and hepatitisB virus Deoxyribonucleicac (hepatitisB virus Deoxyribonucleicac) were evaluated every 3 months. ID, HBV DNA) and the changes in the surface markers of hepatitis B virus (HBV), and explore the virological response, biochemical response, e antigen conversion and drug resistance of LAM, LdT, and ETV single drug treatment.
Result
Part one
After ODN2216 stimulation, the average amount of IFN- alpha secreted by PBMC in group CHB was 31.20 (7.33 to 44.04) pg/ml, and the average of PBMC secreted IFN- alpha in the HBV carrier group was 109.91 (13.74 ~ 240.27) pg/ml, and the average of IFN- alpha in the healthy control group was 107.95 (48.59 to 227.33) and the.CHB patients were significantly lower than those in the healthy control group. The patient group was significantly lower than that of the HBV carrier group (P=0.027).
After poly (I:C) stimulation, the IFN- alpha secretion of PBMC in the CHB group was 37.50 (4.40 to 44.45) pg/ml, and the average of IFN-a in the HBV carrier group was 10.44 (3.46 to 36.08) pg/ml. The average IFN-a volume of the PBMC secretion in the healthy control group was 7.06 (2.89 to 22.80), and there was no significant difference between the three groups.
The second part
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rate of HBV DNA was 68.8%, 63.2%, 80%, and the 81.8%.ALT recurrence rate was 72.2%, 80%, 90.9%, and the serological conversion rate of 83.3%.e antigen was 11.1%, 18.2%, 33.3%, 37.5%., respectively.
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rate of HBV DNA was 90.9%, 90.9%, 66.7%, and the 80.0%.ALT recurrence rate was 63.6%, 88.9%, 100%, and the serological conversion rate of 80.0%.e antigen was 12.5%, 14.3%, 40%, 50.0%., respectively.
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rates of HBV DNA were 84.8%, 83.3%, 100%, 85.7%, and the ALT recurrence rate was 80.6%, 88.9%, 70%, and 75.0%.e antigen serological conversion rates were 0%, 10%, 22.2%, 28.6%., respectively.
4. virology resistance, LAM cumulative virology breakthrough rate was 40.9%, group LdT cumulative virological breakthrough rate was 15.4%, group ETV cumulative virological breakthrough rate was 5.0%.
conclusion
Part one:
The amount of IFN- alpha secreted by dendritic cells from 1.HBV carriers and healthy controls was significantly higher than that of monocytes in PBMC.
Compared with healthy controls, 2.HBV carriers showed no significant difference in secretion of IFN- alpha from dendritic cells in PBMC.
Compared with HBV carriers and healthy controls, the secretion of IFN- alpha in dendritic cells of 3.CHB patients was significantly lower than that in PBMC patients.
In patients with 4.CHB, compared with healthy controls, HBV carriers showed no significant difference in monocyte secretion of IFN- alpha in PBMC.
The second part:
1. virological response, followed up to 144 weeks, there was no significant difference between the three groups of drug treatment,.ETV group received a higher HBV DNA inhibition rate, followed by LAM group and LdT group.
2. in terms of biochemical response, follow-up to 144 weeks, there was no significant difference between the three groups. There was a higher ALT recovery rate in group.LAM, followed by LdT group and ETV group.
The serological conversion of 3.e antigen was followed up to 144 weeks. There was no statistical difference between the three groups. The.LdT group obtained a higher serological conversion rate of e antigen, followed by the ETV and LAM groups.
4. virology resistance, LAM cumulative virology breakthrough rate is much higher than the other two groups, LdT times, ETV group the lowest.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62
本文编号：2146834
[Abstract]:background
Hepatitis B virus (hepatitis B virus, HBV) infection is an infectious disease that seriously affects public health. There are about 93000000 people with HBV infection in China, of which about 30000000 are chronic hepatitis B (chronic hepatitis B, CHB). The immune mechanism of HBV chronic infection is not clear, but more and more studies show that the natural immune system is in it. It plays an important role. Among them, dendritic cell (DC) and mononuclear cells (monocyte, MO) are important natural immune cells, and their role in the chronicity mechanism of HBV infection is worth exploring. Interferon a (interferon-alpha, IFN-a) is an important barrier to the human body against virus infection, and IFN-a in CHB patients is in the presence of IFN-a. Secretory decline is of great clinical significance. More than 95% of IFN-a is secreted by plasma cell like dendritic cells (plasmacytoid dendric cell, pDC). Monocyte (monocyte) is also an important source of secretion of IFN-a, and the presence of IFN-a in mononuclear cells has not been reported.
The most important treatment for CHB patients is antiviral therapy. The advent of nucleoside (acid) (acid) analogues (nucleos (T) ide analogue, NUCs) has opened a new chapter in antiviral therapy. A large number of studies have been made on antiviral therapy for CHB patients at home and abroad, but many of them are derived from phase III clinical test results, and further enrichment of lamivudine (1amivudine) is needed at home. (LAM), telbivudine (LdT) and Cave (entecavir, ETV) in the clinical real world antiviral treatment of long-term follow-up data.
The first part of this study was designed to detect the secretion of IFN-a in peripheral blood mononuclear cells (peripheral blood mononuclear cell, PBMC) in patients with chronic HBV infection, in order to explore whether the function of pDC and mononuclear cells in peripheral blood has defects.
The second part of this study aims to explore the efficacy of long term oral NUCs antiviral therapy in CHB patients in the real world, and compare the difference in efficacy between LAM, LdT and ETV.
Method
The first part of the study prospectively included 16 chronic hepatitis B patients in the Peking Union Medical College Hospital hepatitis clinic from July 2012 to March 2013. 16 cases of HBV carriers were matched with age. The healthy control group was derived from the healthy physical examination center of the hospital and 18 healthy people matched with the age. The blood samples were collected and the PBMC was separated. Not with CPG oligo deoxydeoxynucleotides 2216 (cytosine guanine dinucleotides Oligodeoxy nucleotides, CPG ODN2216), and polyoside acid cytidine acid (Polyinosinic-Polycytidylicacid, poly I:C) stimulated co culture, and the method of enzyme linked immunosorbent assay was used to measure its secretion.
The second part of this study analyzed 87 cases of CHB patients in Peking Union Medical College Hospital hepatitis clinic from June 2008 to April 2013. The longest follow-up time was 192 weeks, and the level of alanine aminotransferase (alanine aminotransferase, ALT) and hepatitisB virus Deoxyribonucleicac (hepatitisB virus Deoxyribonucleicac) were evaluated every 3 months. ID, HBV DNA) and the changes in the surface markers of hepatitis B virus (HBV), and explore the virological response, biochemical response, e antigen conversion and drug resistance of LAM, LdT, and ETV single drug treatment.
Result
Part one
After ODN2216 stimulation, the average amount of IFN- alpha secreted by PBMC in group CHB was 31.20 (7.33 to 44.04) pg/ml, and the average of PBMC secreted IFN- alpha in the HBV carrier group was 109.91 (13.74 ~ 240.27) pg/ml, and the average of IFN- alpha in the healthy control group was 107.95 (48.59 to 227.33) and the.CHB patients were significantly lower than those in the healthy control group. The patient group was significantly lower than that of the HBV carrier group (P=0.027).
After poly (I:C) stimulation, the IFN- alpha secretion of PBMC in the CHB group was 37.50 (4.40 to 44.45) pg/ml, and the average of IFN-a in the HBV carrier group was 10.44 (3.46 to 36.08) pg/ml. The average IFN-a volume of the PBMC secretion in the healthy control group was 7.06 (2.89 to 22.80), and there was no significant difference between the three groups.
The second part
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rate of HBV DNA was 68.8%, 63.2%, 80%, and the 81.8%.ALT recurrence rate was 72.2%, 80%, 90.9%, and the serological conversion rate of 83.3%.e antigen was 11.1%, 18.2%, 33.3%, 37.5%., respectively.
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rate of HBV DNA was 90.9%, 90.9%, 66.7%, and the 80.0%.ALT recurrence rate was 63.6%, 88.9%, 100%, and the serological conversion rate of 80.0%.e antigen was 12.5%, 14.3%, 40%, 50.0%., respectively.
At 24 weeks, 48 weeks, 96 and 144 weeks, the inhibition rates of HBV DNA were 84.8%, 83.3%, 100%, 85.7%, and the ALT recurrence rate was 80.6%, 88.9%, 70%, and 75.0%.e antigen serological conversion rates were 0%, 10%, 22.2%, 28.6%., respectively.
4. virology resistance, LAM cumulative virology breakthrough rate was 40.9%, group LdT cumulative virological breakthrough rate was 15.4%, group ETV cumulative virological breakthrough rate was 5.0%.
conclusion
Part one:
The amount of IFN- alpha secreted by dendritic cells from 1.HBV carriers and healthy controls was significantly higher than that of monocytes in PBMC.
Compared with healthy controls, 2.HBV carriers showed no significant difference in secretion of IFN- alpha from dendritic cells in PBMC.
Compared with HBV carriers and healthy controls, the secretion of IFN- alpha in dendritic cells of 3.CHB patients was significantly lower than that in PBMC patients.
In patients with 4.CHB, compared with healthy controls, HBV carriers showed no significant difference in monocyte secretion of IFN- alpha in PBMC.
The second part:
1. virological response, followed up to 144 weeks, there was no significant difference between the three groups of drug treatment,.ETV group received a higher HBV DNA inhibition rate, followed by LAM group and LdT group.
2. in terms of biochemical response, follow-up to 144 weeks, there was no significant difference between the three groups. There was a higher ALT recovery rate in group.LAM, followed by LdT group and ETV group.
The serological conversion of 3.e antigen was followed up to 144 weeks. There was no statistical difference between the three groups. The.LdT group obtained a higher serological conversion rate of e antigen, followed by the ETV and LAM groups.
4. virology resistance, LAM cumulative virology breakthrough rate is much higher than the other two groups, LdT times, ETV group the lowest.
【学位授予单位】：北京协和医学院
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.62
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