慢性HBV感染者PBMC分泌IFN-α功能初探及慢乙肝患者長期核苷（酸）藥物抗病毒療效
發(fā)布時間：2018-07-26 17:55
【摘要】：背景 乙型肝炎病毒(hepatitis B virus, HBV)感染是一種嚴(yán)重影響公眾健康的傳染病。我國HBV感染者有9300多萬人,其中3000多萬為慢性乙型肝炎(chronic hepatitis B, CHB)患者。目前HBV慢性感染的免疫機(jī)制尚未明確,但越來越多的研究顯示,天然免疫在其中起到重要作用。其中,樹突狀細(xì)胞(dendritic cell, DC)和單核細(xì)胞(monocyte, MO)作為重要的天然免疫細(xì)胞,其在HBV感染慢性化機(jī)制中發(fā)揮的作用值得探究。干擾素a (interferon-alpha, IFN-a)是人體抵御病毒感染的重要屏障,CHB患者體內(nèi)是否存在IFN-a分泌下降具有很重要的臨床意義。人體95%以上的IFN-a由漿細(xì)胞樣樹突狀細(xì)胞(plasmacytoid dendric cell, pDC)分泌。單核細(xì)胞(monocyte)亦是分泌IFN-a的重要來源,單核細(xì)胞是否存在IFN-a的分泌下降未見報道。 CHB患者最主要的治療方法為抗病毒治療。核苷(酸)類似物(nucleos(t)ide analogue, NUCs)藥物的問世掀開了抗病毒治療新的篇章。國內(nèi)外對CHB患者抗病毒治療做了大量研究,但多來源于Ⅲ期臨床試驗(yàn)結(jié)果,國內(nèi)尚需進(jìn)一步豐富拉米夫定(1amivudine, LAM)、替比夫定(telbivudine, LdT)及恩替卡韋(entecavir, ETV)在臨床真實(shí)世界中抗病毒治療效果的長期隨訪資料。研究目的 本研究第一部分旨在檢測慢性HBV感染者外周血單個核細(xì)胞(peripheral blood mononuclear cell, PBMC)分泌IFN-a的情況,以探究其外周血pDC和單核細(xì)胞功能是否存在缺陷。 本研究第二部分旨在探究真實(shí)世界中CHB患者長期口服NUCs藥物抗病毒治療的療效,并比較LAM, LdT,及ETV的療效差異。 方法 本研究第一部分前瞻性納入2012年7月至2013年3月北京協(xié)和醫(yī)院肝炎門診就診的慢性乙型肝炎患者16例,與之年齡相匹配的HBV攜帶者16例。健康對照組來源于北京協(xié)和醫(yī)院健康體檢中心,與之年齡相匹配的健康人18例。采集血樣,分離PBMC,分別與CPG寡聚脫氧核苷酸2216(cytosine guanine dinucleotides oligodeoxy nucleotides, CPG ODN2216)、聚肌苷酸胞苷酸(Polyinosinic-Polycytidylicacid,poly I：C)刺激共培養(yǎng),并用酶聯(lián)免疫吸附法(enzyme linked immunosorbent assay,Elisa)的方法測量其分泌IFN-a量。 本研究第二部分回顧性分析2008年6月至2013年4月于北京協(xié)和醫(yī)院肝炎門診就診的CHB患者87例,最長隨訪時間為192周,每3個月評估其丙氨酸氨基轉(zhuǎn)氨酶(alanine aminotransferase, ALT)水平、乙型肝炎病毒脫氧核糖核酸(hepatitisB virus Deoxyribonucleicacid, HBV DNA)和乙型肝炎病毒表面標(biāo)志物的變化情況,探究LAM,LdT,及ETV單藥治療的病毒學(xué)應(yīng)答,生化學(xué)應(yīng)答,e抗原轉(zhuǎn)換及耐藥情況。 結(jié)果 第一部分 經(jīng)ODN2216刺激后,CHB組患者PBMC分泌IFN-α量平均為31.20(7.33～44.04)pg/ml, HBV攜帶者組PBMC分泌IFN-α量平均為109.91(13.74～240.27)pg/ml,健康對照組PBMC分泌IFN-α量平均為107.95(48.59～227.33)。CHB患者組明顯低于健康對照組,具有統(tǒng)計(jì)學(xué)差異(P=0.007)。CHB患者組明顯低于HBV攜帶者組,具有統(tǒng)計(jì)學(xué)差異(P=0.027) 經(jīng)poly(I：C)刺激后,CHB患者組PBMC分泌IFN-α量平均為37.50(4.40～44.45)pg/ml,HBV攜帶者組PBMC分泌IFN-a量平均為10.44(3.46～36.08)pg/ml,健康對照組PBMC分泌IFN-a量平均為7.06(2.89～22.80)pg/ml,三組之間無明顯差異(P=0.427)。 第二部分 1.LAM組患者治療24周、48周、96周和144周HBV DNA抑制率為68.8%,63.2%,80.0%,81.8%。ALT復(fù)常率為72.2%,80.0%,90.9%,83.3%。e抗原血清學(xué)轉(zhuǎn)換率分別為11.1%,18.2%,33.3%,37.5%。 2.LdT組患者治療24周、48周、96周和144周HBV DNA抑制率為90.9%,90.9%,66.7%,80.0%。ALT復(fù)常率為63.6%,88.9%,100.0%,80.0%。e抗原血清學(xué)轉(zhuǎn)換率分別為12.5%,14.3%,40.0%,50.0%。 3.ETV組患者治療24周、48周、96周和144周HBV DNA抑制率為84.8%,83.3%,100%,85.7%,ALT復(fù)常率為80.6%,88.9%,70.0%,75.0%。e抗原血清學(xué)轉(zhuǎn)換率分別為0%,10.0%,22.2%,28.6%。 4.病毒學(xué)耐藥方面,LAM累積病毒學(xué)突破率為40.9%,LdT組累積病毒學(xué)突破率為15.4%,ETV組累積病毒學(xué)突破率為5.0%。 結(jié)論 第一部分： 1.HBV攜帶者和健康對照PBMC中樹突狀細(xì)胞分泌IFN-α的量顯著高于單核細(xì)胞。 2.HBV攜帶者與健康對照相比,PBMC中樹突細(xì)胞分泌IFN-α無顯著差異。 3.CHB患者相比HBV攜帶者、健康對照,PBMC中樹突狀細(xì)胞分泌IFN-α明顯下降。 4.CHB患者、HBV攜帶者相比健康對照,PBMC中單核細(xì)胞分泌IFN-α無顯著差異。 第二部分： 1.病毒學(xué)應(yīng)答方面,隨訪至144周,三組藥物治療無統(tǒng)計(jì)學(xué)差異。ETV組獲得較高的HBV DNA抑制率,其次為LAM組和LdT組。 2.生化學(xué)應(yīng)答方面,隨訪至144周,三組藥物治療無統(tǒng)計(jì)學(xué)差異。LAM組獲得了較高的ALT復(fù)常率,其次為LdT組和ETV組。 3.e抗原血清學(xué)轉(zhuǎn)換方面,隨訪至144周,三組藥物治療無統(tǒng)計(jì)學(xué)差異。LdT組獲得了較高的e抗原血清學(xué)轉(zhuǎn)換率,其次為ETV組和LAM組。 4.病毒學(xué)耐藥方面,LAM累積病毒學(xué)突破率遠(yuǎn)遠(yuǎn)高于其余兩組,LdT次之,ETV組最低。
[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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R512.62
本文編號：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.
【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2013
【分類號】：R512.62
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