胆汁酸升高的慢性乙型肝炎患者T细胞免疫衰老及肝细胞癌风险升高

发布时间：2018-01-15 22:18

本文关键词：胆汁酸升高的慢性乙型肝炎患者T细胞免疫衰老及肝细胞癌风险升高　出处：《第三军医大学》2016年博士论文　论文类型：学位论文

【摘要】：随着年龄的增加,天然免疫与获得性免疫系统功能进行性的退化,最终导致免疫力的降低,这个现象通常被称为免疫衰老。免疫衰老通常被认为与老年人对疫苗免疫反应减低、易于遭受病毒或细菌感染、易于发生癌症相关。作为获得性免疫系统的主要组成,免疫衰老过程中T细胞的功能退化有着最多的关注。T细胞免疫衰老通常表型为T细胞免疫系统多个层面上的退行性变化,其中包括:naive T细胞数量下降,TCR(T-cell Receptor,T细胞受体)repertoire多样性降低,T细胞端粒缩短,memory T细胞功能下降等。目前研究结果表明慢性病原体感染对于机体免疫系统有着负面的影响。在本研究中,我们分析了是否CHB(Chronic Hepatitis B,慢性乙型肝炎)患者其T细胞免疫系统受损,同时我们也分析了是否CHB患者T细胞免疫系统受损与其发生HCC(Hepatocellular Carcinoma,肝细胞癌)相关。我们比较了24例CHB患者和15例健康人其naive T细胞库的TCR repertoire。与以前研究报道结果一致,健康人40岁以前其naive T细胞库的TCR repertoire多样性没有明显降低保持在高水平。与健康人不同,CHB患者40岁以前其naive T细胞库的TCR repertoire多样性随着年龄的增加显著降低。考虑到这24例CHB患者都是通过垂直传播感染的HBV(Hepatitis B Virus,乙型肝炎病毒),我们认为其naive T细胞库的TCR repertoire多样性的降低是由于HBV病毒与T细胞免疫系统长期作用的结果而不是由年龄增加造成的。和健康人naive T细胞库一样,没有表达特定TCR克隆型的T细胞克隆在naive T细胞库中占据主导地位,这表明CHB患者其naive T细胞库的TCR repertoire多样性的降低不是由某些特定TCR克隆型的T细胞克隆优势性的扩增导致,而很可能反映了CHB患者其naive T细胞库T细胞克隆均匀性的丢失。我们这部分研究的发现表明慢性CHB感染能够加速CHB患者的T细胞免疫衰老。Naive T细胞库的TCR repertoire多样性的降低意味着CHB患者外周T细胞库的维持失衡。成年人外周T细胞库的维持主要通过T细胞的缓慢增殖来实现。首先,我们分析了146例CHB患者及53例健康人外周T细胞的增殖情况。与健康人相比,CHB患者外周t细胞增殖水平显著升高。部分chb患者外周t细胞的增殖水平升高到了hiv-1(humanimmunodeficiencyvirustype1,人免疫缺陷病毒1型)感染的水平。chb患者血清tba(totalbildacid,总胆汁酸)水平与其外周t细胞增殖水平升高显著相关,这主要体现在:从定性的角度分析,血清tba升高的chb患者其所有t细胞亚群(包括naivet细胞和memoryt细胞)的增殖频率相对于血清tba正常的chb患者及健康人都显著升高;从定量的角度分析,血清tba升高的chb患者其外周t细胞的增殖频率与其血清tba显著正相关。我们也分析了chb患者其外周t细胞增殖水平升高与其它chb相关的临床指标的相关性,没有发现类似的显著相关性。随后,我们分析了11例血清tba持续升高的chb患者两个不同时间点的t细胞增殖水平,发现其t细胞在前后两个时间点都保持高水平增殖。这表明对于血清tba持续升高的chb患者,其外周t细胞很可能一直保持在过度增殖状态。进一步,我们分析了12例血清tba升高的chb患者其外周t细胞的活化情况,同时比较了这12例chb患者与3例健康人外周增殖的t细胞与静息的t细胞其tcrvβ家族的分布。血清tba升高的chb患者其增殖的t细胞上调cd38表达,显示其免疫活化。尽管增殖水平差异巨大,血清tba升高的chb患者与健康人其增殖的t细胞与静息的t细胞的tcrvβ家族的分布都高度类似。这表明抗原非特异性的旁路活化(by-standeractivation)机制主导了血清tba升高的chb患者其外周t细胞增殖水平的升高。最后,我们分析了5例健康人、21例有4年以上跟踪的chb患者、4例hbv相关的hcc患者其naivet细胞的端粒长度。相对于健康人及跟踪期内血清tba无升高或者仅一过性升高的chb患者,跟踪期内血清tba持续升高的chb患者其naivet细胞的端粒长度显著缩短,缩短到了与hbv相关的hcc患者类似的水平。我们这部分的研究表明chb患者的t细胞免疫衰老与chb患者血清tba持续升高有紧密的相关性,很可能由血清tba持续升高的chb患者其外周t细胞长期保持异常活化增殖导致。为了研究血清tba持续升高对于chb患者发生hcc的影响,我们做了一个回顾性的观察性的队列研究。回顾性研究的数据来源于hbs(hepatitisbiobankatsouthwesthospital,西南医院肝炎样本库)项目2001年至2014年的数据,该样本库包含了到西南医院传染科就诊的患者其人口统计学方面及临床指标方面的一系列记录。按照我们的筛选标准,一共有3021位chb患者进入最终队列,其中3018(99.%)位在跟踪期间都有定期的通过干扰素或者核苷类似物的抗病毒治疗。我们利用coxproportionalhazardmodels、kaplan-meieranalyses、以及propensityscoreanalyses来分析血清TBA持续升高与CHB患者发生HCC的相关性。我们的分析选择了性别、年龄、肝硬化、血清ALT(Alanine Aminotransferase,丙氨酸氨基转移酶)、血清HBV DNA、以及血清HBe Ag(Hepatitis B e Antigen,乙型肝炎e抗原)这几个公认的CHB患者HCC发生的危险因素与血清TBA一起作为协变量(covariate)。CHB患者跟踪期间血清TBA升高的持续程度由其血清TBA升高的跟踪时间占其总跟踪时间的比值决定。根据这个比值,我们把CHB患者其跟踪期间血清TBA长期变化模式按照升高的持续程度分为3群:血清TBA无升高或一过性升高、血清TBA中等水平的持续升高、血清TBA高水平的持续升高。回顾性研究总跟踪期为20,813.7 person-years,一共有81例CHB患者跟踪期内发生了HCC。在3021例队列CHB患者中,血清TBA无升高或一过性升高的CHB患者为2139(70.8%)例,血清TBA中等水平的持续升高的CHB患者为520(17.2%)例,血清TBA高水平的持续升高的CHB患者为362(12.0%)例。整个跟踪期内,血清TBA长期变化模式为无升高或一过性升高、中等水平的持续升高、及高水平的持续升高的CHB患者其HCC累积发病率分别为2.2%、6.3%、及19.3%。相对于血清TBA长期变化模式为无升高或一过性升高的CHB患者,血清TBA长期变化模式为中等水平的持续升高、及高水平的持续升高的CHB患者其多因素校正(multivariate adjusted)之后的HRs(Hazard Ratios,风险比)及95%CI(Confidence Interval,置信区间)分别为1.70(0.88-3.26)、及3.08(1.69-5.60)。基于该HRs值计算出来血清TBA长期变化模式为中等水平的持续升高、及高水平的持续升高对于HCC发生的PARs(Population Attributable Risks,人群归因危险度)分别为7.8%、及29.0%。基于propensity score models的分析也验证了CHB患者其跟踪期间血清TBA升高的持续程度与HCC发生的这种剂量-反应关系(Dose-response Relationship)。我们这部分的研究表明,血清TBA的持续升高是定期接受抗病毒治疗的CHB患者人群发生HCC的主要的、独立的风险因素。综上所述,我们的研究表明血清TBA的持续升高的CHB患者其HCC发生的风险升高,这群CHB患者升高的HCC风险很可能是由其T细胞免疫衰老导致。实际临床应用上,我们的发现表明对血清TBA水平的持续升高的CHB患者执行定期的HCC筛查能够保证在该HCC高风险患者群体及时地发现早期HCC,有利于后期的治疗。理论层面上,我们的发现对于从免疫学的角度来理解HCC的发生提供了一个很好的切入点。
[Abstract]:With the increase of age, innate and acquired immune system functional degradation, resulting in decreased immunity, this phenomenon is often called immunosenescence. Immunosenescence is usually considered with the elderly on immune responses to vaccines reduced, prone to viral or bacterial infection, prone to cancer. As a major component of gain of the immune system, T cell immune function degradation during senescence is concerned.T cell immune senescence most often phenotypes of degenerative changes, multiple levels of T cell immune system including: the number of naive T cells decreased, TCR (T-cell Receptor, T cell receptor) repertoire diversity decreased telomere T cell shortening, function of memory T cells decreased. The results of the present study showed that chronic infection has a negative impact on the immune system. In this study, we analyzed is No CHB (Chronic Hepatitis B, chronic hepatitis B) in patients with impaired T cell immune system at the same time, we also analyzed whether CHB T cells in patients with impaired immune systems and HCC (Hepatocellular Carcinoma, HCC). We compared 24 patients with CHB and 15 healthy people in the naive T Library of TCR cells repertoire. reported results consistent with previous studies, 40 healthy people before the age of the naive T cell library TCR repertoire diversity did not significantly decrease remained at a high level. Different from healthy people, patients aged 40 years before CHB naive T cell library TCR repertoire diversity increased with age significantly reduced. Considering the 24 cases of CHB patients are infected by vertical transmission of HBV (Hepatitis B Virus, hepatitis B virus), we believe that to reduce the naive T cell library TCR repertoire diversity is due to the HBV virus and T cell immune system The result of long-term effect and not by age increases. Like healthy human naive T cell library, T cell clones did not express specific TCR clone type occupies the dominant position in the naive T cell library, which indicates that CHB patients with naive T TCR repertoire cell library diversity reduction was not caused by T cells the advantages of cloning some specific TCR amplification and cloning of the type, is likely to reflect the loss of CHB in the naive of patients with T cell T cell clone library uniformity. We found this part of the study showed that chronic CHB infection can accelerate T cell immunity in patients with CHB.Naive T cells senescence library TCR repertoire diversity reduction means maintain the imbalance of T cell library CHB patients. Adult peripheral T cells maintain the library mainly through the slow proliferation of T cells to achieve. Firstly, we analyzed 146 cases of CHB patients and 53 cases of healthy human peripheral T cells The proliferation of CHB. Compared with healthy people, patients with peripheral T cell proliferation was significantly increased. CHB patients with peripheral T cell proliferation level was increased to HIV-1 (humanimmunodeficiencyvirustype1, human immunodeficiency virus type 1 infection) level of serum.Chb in patients with TBA (total bile acid totalbildacid) and elevated levels of peripheral T cells the proliferation was significantly related, which is mainly reflected in: analysis from the qualitative point of view, the level of serum TBA in CHB patients with all its subsets of T cells (including naivet cells and memoryt cells) proliferation frequency with serum TBA in CHB patients and normal healthy people was significantly increased; the analysis from the quantitative point of view is significantly related to serum TBA increased in CHB patients with peripheral T cell proliferation frequency and serum TBA. We also analyzed the correlation between increased in patients with CHB peripheral T cell proliferation level and other clinical indicators related to CHB, There was no significant correlation between similar findings. Then, we analyzed the proliferation level of T cells in 11 cases of CHB patients with persistently elevated serum level of TBA in two different time points, and it is found that the T cells maintain a high level of proliferation at two time points before and after. This showed that the serum TBA elevated CHB patients, peripheral T cells it may keep in excessive proliferation. Further, we analyzed the activation of 12 cases of elevated serum TBA CHB in patients with peripheral T cells, and compare the distribution of T cells and the rest 12 cases of CHB patients and 3 healthy people and Zhou Zengzhi's T cell TCRV beta family. The expression of serum TBA increased the proliferation of T cells in CHB patients showed upregulation of CD38, the proliferation of immune activation. Although there is a huge difference, the distribution of TCRV beta family T cells and resting CHB patients and healthy human serum TBA elevated the proliferation of T cells are highly similar This shows that the bypass. Antigen nonspecific activation (by-standeractivation) mechanism dominates the increase of serum TBA in patients with CHB elevated peripheral T cell proliferation level. Finally, we analyzed 5 cases of healthy people, 21 cases of more than 4 years follow-up of CHB patients, the telomere length in 4 cases of HBV associated HCC in naivet cells compared to healthy people. And the tracking period of serum TBA increased no or only a transient increase in CHB patients, the tracking period of persistently elevated serum level of TBA CHB in naivet cells significantly shortened telomere length, reduced to a similar HBV related with HCC level. This part of our study showed that patients with CHB t cellular senescence and CHB in serum of patients with persistently elevated TBA is associated with CHB in peripheral T cells, possibly by increasing serum TBA long-term abnormal activation and proliferation cause. In order to study the serum TBA increased The effect of CHB in patients with HCC, we conducted a retrospective observational cohort study. A retrospective study of data from the HBS (hepatitisbiobankatsouthwesthospital, Southwest Hospital Sample Library) project in 2001 to 2014 data, the sample library contains a set of records to the Southwest Hospital Department of infectious disease patients and its demographics clinical indicators. According to our selection criteria, a total of 3021 patients with CHB into the final queue, of which 3018 (99%) in the tracking period regularly through antiviral therapy of interferon or nucleoside analogs. We use coxproportionalhazardmodels, kaplan-meieranalyses, and propensityscoreanalyses to analyze the relationship between the serum levels of TBA increased in patients with CHB HCC the analysis. We chose the gender, age, liver cirrhosis, serum ALT (Alanine Aminot Ransferase, alanine aminotransferase), serum HBV DNA, serum HBe and Ag (Hepatitis B e Antigen, hepatitis B e antigen) the risk factors of several recognized CHB HCC patients with serum TBA as covariates (covariate) patients with.CHB ratio during continuous tracking of the level of serum TBA by the serum TBA the tracking time of the total time tracking decision. According to this ratio, we put the CHB in tracking mode according to long-term changes in serum TBA during sustained elevated levels of serum TBA were divided into 3 groups: no increase or a transient increase in serum TBA, medium level continues to rise, high serum levels of TBA were rising. Study on the total follow-up period was 20813.7 person-years, a total of 81 cases of CHB patients with follow-up period occurred in 3021 cases of HCC. queue in CHB patients, serum TBA increased or a transient increase in CHB patients was 2139 (70.8%) Patients, serum TBA level increased in moderate CHB patients was 520 (17.2% cases), high serum levels of TBA increased in CHB patients was 362 (12%) cases. The follow-up period, serum TBA long-term variation pattern with no increase or a transient increase, the medium level continues to rise, and a high level the rising of CHB patients the cumulative incidence of HCC were 2.2%, 6.3%, and 19.3%. relative to the long-term changes in serum TBA pattern with no increase or a transient increase in CHB patients, serum TBA increased for long-term changes in mode of moderate levels of correction and high level increased in CHB patients (multi factor multivariate adjusted HRs (Hazard) after Ratios, the risk ratio) and 95%CI (Confidence Interval CI) were 1.70 (0.88-3.26), and 3.08 (1.69-5.60). The HRs value calculated from the increasing serum TBA of long term changes for intermediate level based on model, The high level and continues to rise for the occurrence of HCC PARs (Population Attributable Risks, the population attributable risk was 7.8%), and 29.0%. analysis of propensity score based on models also verified the relationship between patients with CHB and HCC during the tracking duration of elevated serum TBA occurred in this kind of dose response (Dose-response Relationship) that we. Some studies show that serum TBA increased regularly receive antiviral treatment of patients with CHB who have HCC the main independent risk factors. In summary, our study showed that the increased risk of sustained high level of serum TBA in CHB patients HCC, this group of patients with CHB increased the risk of HCC is likely to be caused by the T cell immunosenescence. Clinical application, we found that the level of serum TBA increased in patients with CHB HCC to ensure the regular screening The HCC high-risk group can find early HCC in time, which is conducive to the later treatment. In theory, our findings provide a good breakthrough point for understanding the occurrence of HCC from the perspective of immunology.

【学位授予单位】：第三军医大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R735.7;R512.62

【相似文献】