CD4^+CXCR5^+ T cells activate CD27^+IgG^+ B cells via IL-21 in patients with hepatitis C virus infection

BACKGROUND： Chronic hepatitis C virus （HCV） infection causes the skewing and activation of B cell subsets, but the characteristics of IgG＋ B cells in patients with chronic hepa- titis C （CHC） infection have not been thoroughly elucidated. CD4＋CXCR5＋ follicular helper T （Tfh） cells, via interleukin （IL）-21 secretion, activate B cells. However, the role of CD4＋CXCR5＋ T cells in the activation ofIgG＋ B cells in CHC patients is not clear. METHODS： The frequency of IgG＋ B cells, including CD27-IgG＋ B and CD27＋IgG＋ B cells, the expression of the activation markers （CD86 and CD95） in IgG＋ B cells, and the percentage of circu- lating CD4＋CXCR5＋ T cells were detected by flow cytometry in CHC patients （n=70） and healthy controls （n=25）. The con- centrations of serum IL-21 were analyzed using ELISA. The role of CD4＋CXCR5＋ T cells in the activation of IgG＋ B cells was investigated using a co-culture system. RESULTS： A significantly lower proportion of CD27＋IgG＋ B cells with increased expression of CD86 and CD95 was observed in CHC patients. The expression of CD95 was negatively correlated with the percentage of CD27＋IgG＋ B cells, and it contributed to CD27＋IgG＋ B cell apoptosis. Circulating CD4＋CXCR5＋ T cells and serum IL-21 were significantly increased in CHC patients. Moreover, circulating CD4＋CXCR5＋ T cells from CHC patients induced higher expressions of CD86 and CD95 in CD27＋IgG＋ B cells in a co-culture system; the blockade of the IL-21 decreased the expression levels of CD86 and CD95 in CD27＋IgG＋ B cells.CONCLUSIONS： HCV infection increased the frequency of CD4＋CXCR5＋ T cells and decreased the frequency of CD27＋IgG＋ B cells. CD4＋CXCR5＋ T cells activated CD27＋IgG＋ B cells via the secretion of IL-21.

CD4+ T cells and natural killer cells: Biomarkers for hepatic fibrosis in human immunodeficiency virus/hepatitis C virus-coinfected patients

AIM To characterize peripheral blood natural killer(NK) cells phenotypes by flow cytometry as potential biomarker of liver fibrosis in human immunodeficiency virus(HIV)/hepatitis C virus(HCV) coinfected patients.METHODS Peripheral mononuclear cells from 24 HIV/HCV(HBVnegative) coinfected and 5 HIV/HCV/HBV seronegative individuals were evaluated. HIV/HCV coinfected patients were divided in to groups: G1, patients with METAVIR F0-F2 and G2, patients with METAVIR F3-F4. NK surface cell staining was performed with: AntiCD3(APC/Cy7), anti-CD56(PE/Cy5), anti-CD57(APC), anti-CD25(PE), anti-CD69(FITC), anti-NKp30(PE), antiNKp46(PE/Cy7), anti-NKG2D(APC), anti-DNAM(FITC); anti-CD62L(PE/Cy7), anti-CCR7(PE), anti-TRAIL(PE), anti-Fas L(PE), anti CD94(FITC). Flow cytometry data acquisition was performed on BD FACSCanto, analyzed using Flow Jo software. Frequency of fluorescence was analyzed for all single markers. Clinical records were reviewed, and epidemiological and clinical data were obtained.RESULTS Samples from 11 patients were included in G1 and from 13 in G2. All patients were on ARV, with undetectable HIV viral load. Liver fibrosis was evaluated by transient elastography in 90% of the patients and with biopsy in 10% of the patients. Mean HCV viral load was(6.18 ± 0.7 log10). Even though, no major significant differences were observed between G1 and G2 regarding NK surface markers, it was found that patients with higher liver fibrosis presented statistically lower percentage of NK cells than individual with low to mild fibrosis and healthy controls(G2: 5.4% ± 2.3%, G1: 12.6% ± 8.2%, P = 0.002 and healthy controls 12.2% ± 2.7%, P = 0.008). It was also found that individuals with higher liver fibrosis presented lower CD4 LT count than those from G1(G2: 521 ± 312 cells/μL, G1: 770 ± 205 cells/μL; P = 0.035).CONCLUSION Higher levels of liver fibrosis were associated with lower percentage of NK cells and LTCD4+ count; and they may serve as noninvasive biomarkers of liver damage.

CD4+ T cell responses in hepatitis C virus infection

Hepatitis C virus (HCV) infection is a major cause of liver damage, with virus-induced end-stage disease such as liver cirrhosis and hepatocellular carcinoma resulting in a high rate of morbidity and mortality worldwide. Evidence that CD4+ T cell responses to HCV play an important role in the outcome of acute infection has been shown in several studies. However, the mechanisms behind viral persistence and the failure of CD4+ T cell responses to contain virus are poorly understood. During chronic HCV infection, HCV-specific CD4+ T cell responses are rela- tively weak or absent whereas in resolved infection these responses are vigorous and multispecific. Persons with a T-helper type I profile, which promotes cellular effec- tor mechanisms are thought to be more likely to experi- ence viral clearance, but the overall role of these cells in the immunopathogenesis of chronic liver disease is not known. To define this, much more data is required on the function and specificity of virus-specific CD4+ T cells, especially in the early phases of acute disease and in the liver during chronic infection. The role and possible mechanisms of action of CD4+ T cell responses in deter- mining the outcome of acute and chronic HCV infection will be discussed in this review.

Natural course of chronic hepatitis B is characterized by changing patterns of programmed death type-1 of CD8-positive T cells

AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection were enrolled in this study.PD-1 expression in total T cells was detected by flow cytometry.Levels of total CD8+T cell responses and proliferation in relation to PD-1 expression levels were analyzed with intracellular staining and PD-1/ PD-L1 blockage. RESULTS:The PD-1 expression in T cells was dynamically changed during the natural course of chronic HBV infection,did not significantly increase in the immune tolerance phase,and returned to normal in the inactive virus carrier stage.Blockage of the PD-1/PD-L1 pathway could not affect the T-cell response in the immune tolerance and inactive virus carrier stages of chronic HBV infection.However,it could significantly restore the T-cell response in the immune clearance stage of chronic HBV infection.Furthermore,the PD-1 expression level in T cells was associated with the alanine aminotransferase level during the immune clearance stage of chronic HBV infection. CONCLUSION:The PD-l/PD-L1 pathway plays a different role in T-cell response during the natural course of chronic HBV infection.

Ribavirin and IFN-α combination therapy induces CD4+ T-cell proliferation and Th1 cytokine secretion in patients with chronic hepatitis B

AIM: To investigate the anti-viral mechanism of combination therapy of interferon (IFN)-α and ribavirin in patients with chronic hepatitis B. METHODS: Twenty patients were assigned to receive either IFN-α plus ribavirin (group A,n = 14) or no treatment as a control (group B,n = 6). Patients were analyzed for T-cell proliferative responses specific for hepatitis B virus (HBV)-antigen and cytokine production by peripheral blood mononuclear cells (PBMCs). RESULTS: Combination therapy induced HBV-antigen specific CD4+ T-cell proliferative responses in four patients (28.6%). Production of high levels of HBV-specific IFN-γ,tumor necrosis factor (TNF)-α,interleukin (IL)-12 by PBMCs was found in five patients (35.7%),who showed significantly lower HBV DNA levels in serum at 12 mo after treatment ended (P = 0.038) and at 24 mo of follow-up (P = 0.004) than those without high levels of cytokine production. CONCLUSION: HBV-antigen specific CD4+ T cells may directly control HBV replication and secretion of anti-viral T helper 1 (Th1) cytokines by PBMCs during combination therapy of chronic hepatitis B with ribavirin and IFN-α.

Effect of macrophage polarization regulated by miR-29b,B7H3 on CD4^(+)T cell differentiation in asthma

Objective:To explore the mechanism that miR-29b and B7H3 regulate the polarization of macrophages and thus affect the differentiation of CD4^(+)T.Methods:1.PBMC was extracted from peripheral blood mononuclear cells of children with asthma and normal children in the affiliated Children's Hospital of Soochow University,and RNA was extracted and reverse transcribed.The expression of miR-29b and B7H3mRNA was determined by real-time quantitative polymerase chain reaction(Q-PCR).The family history of asthma and history of allergic diseases were collected.2.THP-1 cells were induced into macrophages,miR-29b interference,miR-29b overexpression and normal control were induced by LV526,LV527 and NC virus infection.After 24 hours of culture,the cells were collected to detect the expression of STAT3 and B7H3 genes and proteins.3.It was verified that STAT3 was the target gene of miR-29b:after inoculating THP-1 cells and culturing with PMA with final concentration of 50ng/ml for 6 hours,the macrophages without PMA were cultured for 24 hours,then the macrophages infected by LV528,LV529 and NC virus were induced to form miR-29b interference,miR29b overexpression and normal control group.Luciferase analysis was performed at 48 hours to verify that STAT3 was the target gene of miR-29b.STAT3-3'UTR luciferase reporter gene plasmids were constructed and divided into three groups:"miR-29b+STAT3-3'UTR","miR-29b+STAT3-mut-3'UTR"and"miR-29b+luciferase empty load".4.Macrophages with different treatments were co-cultured with initial T cells for 3 days.The relative expressions of T-bet,GATA3 and ROR-γt were detected by Q-PCR.Result:1.The incidence of allergic disease in the acute attack group(68%)was higher than that in the other two groups(34.8%,33.3%),and the family history of asthma in the normal group(0%)was much lower than that in the other two groups(52%,60.9%).The difference was statistically significant(P<0.05).2.The expression of B7H3 in PBMC in acute attack group was higher than that in non-acute attack group and normal group.The expression of miR-29b in PBMC in normal group was significantly higher than that in non-acute attack group and acute attack group(P<0.0001).The expression of miR-29b in non-acute attack group was significantly higher than that in acute attack group(P=0.007).3.After silencing the expression of miR-29b,IL-4Rα,IL-4,IL-5,IL-13 and CD206 of macrophages increased significantly,while IFN-γdecreased,suggesting that miR-29b can promote the polarization of macrophages to M2.4.The overexpression of miR-29b,STAT3 and B7H3 gene and protein level in macrophages decreased,while the increase of miR-29b,STAT3 and B7H3 gene and protein expression was inhibited.5.There was a significant positive correlation between the expression of STAT3 and B7H3mRNA in macrophages(r=0.9737,P<0.0001).6.STAT3 is the target gene of miR-29b.7.Co-culture of macrophages with CD4^(+)T cells can promote the differentiation of primary T cells,namely Th 0 cells,into Th2,and the promoting effect of macrophages with downregulation of miR-29b is more obvious.Conclusion:The expression of miR-29b in PBMC of children with asthma is lower than that of normal children,while the expression of B7H3 is higher than that of normal children.It is speculated that miR-29b has a protective effect on children with asthma,while B7H3 aggravates the inflammatory response.Down-regulation of miR-29b,in macrophages can promote macrophages to M2 polarization,increase the expression of B7H3 and STAT3 in macrophages,make Th0 cells differentiate into Th2 cells,and aggravate the inflammatory response in patients with asthma.

Parallel decline of CD8+CD38+ lymphocytes and viremia in treated hepatitis B patients

AIM:To assess the peripheral T lymphocyte subsets in chronic hepatitis B virus(HBV) infection,and their dynamics in response to adefovir dipivoxil monotherapy.METHODS:Proportions and absolute counts of peripheral natural killer cells,B cells,CD8+,CD4+,CD8+ CD38+,CD8+CD28+ and CD4+CD28+ T cells were determined using three-color flow cytometry in chronic hepatitis B patients(n = 35),HBV carriers(n = 25) and healthy controls(n = 35).Adefovir dipivoxil was initiated in 17 chronic hepatitis B patients who were regularly followed for 72 wk,during which period the T cell subsets and serum viral load were measured at each follow-up point.RESULTS:The peripheral CD4+ T cell counts and CD8+ T cell counts decreased in chronic HBV infection.In chronic hepatitis B patients,proportions of CD8+CD38+ T cells were 62.0% ± 14.7%,much higher than those of HBV carriers and healthy con-trols.In the 13 hepatitis B patients who were treated and responded to adefovir dipivoxil,proportions of CD8+CD38+ T cells decreased from 53.9% ± 18.4% pre-therapy to 20.1% ± 11.3% by week 72(P < 0.001),concomitant with viral load decline(HBV DNA fell from 7.31 to 3 log copies/mL).CD8+ T cell counts also underwent an average increase of 218 cells/μL by the end of 72-wk treatment.In those who failed the therapy,the CD8+CD38+ T cell population had more fluctuations.CONCLUSION:CD8+ T cells abnormally activated in chronic HBV infection can be partially reversed by antiviral therapy.HBV-associated immune activation may be a crucial part of the pathogenesis and a promising target of treatment.

从HCV蛋白一级结构预测其C,E,NS5区的CD_4^+ T细胞抗原位点

我们根据ＣＤ４＋Ｔ细胞识别抗原位点的物理化学和生物学特征，设计了一个具有查找两亲性螺旋状结构（ａｍｐｈｉｐａｔｈｉｃｈｅｌｉｘｓｔｒｕｃｔｕｒｅ）肽段功能的计算机程序。用该程序对ＨＣＶ－１型病毒Ｃ、Ｅ（Ｅ１、Ｅ２／ＮＳ１）、ＮＳ５蛋白一级结构进行分析，发现这些蛋白区存在ＣＤ４＋Ｔ细胞识别位点。此结果支持了ＣＤ４＋Ｔ细胞对ＨＣＶＣ，Ｅ，ＮＳ５区可发生增殖反应的结论。提示该程序可作为一种预测ＣＤ４＋Ｔ细胞识别ＨＣＶ抗原位点的方法。

Viral infection parameters not nucleoside analogue itself correlates with host immunity in nucleoside analogue therapy for chronic hepatitis B

AIM: To determine the relationship between host immunity and the characteristics of viral infection or nucleoside analogues (NAs) themselves in patients with chronic hepatitis B (CHB) receiving NA therapy.

CD24 aggravates acute liver injury in autoimmune hepatitis by promoting IFN-γ production by CD4^(+) T cells

The T-cell-mediated immune response is implicated in many clinical hepatic injuries, such as autoimmune hepatitis and acute virus hepatitis. CD24 is widely expressed by different immune cells and plays an important role in the pathogenesis of many autoimmune diseases. However, the role of CD24 in T-cell-mediated liver injury has not been elucidated until now. Here we showed that CD24 deficiency protects mice from concanavalin A (ConA)-induced fulminant liver injury by reducing serum interferon-γ (IFN-γ) levels. CD24 expression by hepatic T cells was markedly increased following ConA challenge. Moreover, decreased IFN-γ production by hepatic CD4^(+) T cells in CD24-deficient mice was detected, which was correlated with downregulated phosphorylation of STAT1 in hepatic tissue. In vitro experiments also supported the conclusion that CD24 deficiency impaired IFN-γ production by CD4^(+) T cells following ConA, CD3/CD28 and phorbol myristate acetate/ionomycin stimulation. Our study suggests that CD24 deficiency confers hepatoprotection by decreasing CD4^(+) T-cell-dependent IFN-γ production in vivo, which suggests that CD24 might be a potential target molecule for reducing clinical hepatitis.

TIM-3和CTLA-4基因多态性在HBV感染和HCC中的作用

目的探讨慢性HBV感染者和肝细胞癌患者细胞毒性T淋巴细胞相关抗原-4(cytotoxic T lymphocyte associated antigen-4,CTLA-4)和T细胞免疫球蛋白及黏蛋白分子-3(T cell immunoglobulin and mucin domain-3,TIM-3)的基因多态性分布,以及二者的联合作用。方法通过病例对照研究方法,提取439例慢性HBV感染者(无症状携带者48例,慢性肝炎154例,肝硬化134例和肝癌103例)和220例健康对照者的基因组DNA,限制性片段长度多态性聚合酶链反应(PCR-RFLP)技术检测所有患者DNA的CTLA-4+49 A/G和TIM-3-1516 G/T基因型分布频率,采用χ^(2)检验分析基因型频率、等位基因频率在HBV感染组与对照组的分布情况以及在肝癌组与非肝癌组的分布情况。结果HBV感染组的CTLA-4+49含A基因型(GA+AA)频率高于对照组(P<0.001,OR=1.924)。HBV感染组的TIM-3-1516含T基因型(GT+TT)频率高于对照组(P=0.002,OR=2.263)。CTLA-4+49 GG/TIM-3-1516 GG基因型组合在HBV感染组中的分布频率低于对照组(P<0.001,OR=0.242)。非肝癌组(无症状携带者、慢性肝炎和肝硬化患者)的CTLA-4+49 GA和AA基因型频率高于肝癌组(分别为P<0.001,OR=2.433和P=0.003,OR=2.798)。非肝癌组的TIM-3-1516 GG基因型频率高于肝癌组(P=0.042,OR=1.725)。CTLA-4+49 GA/TIM-3-1516 GG和CTLA-4+49 AA/TIM-3-1516 GG基因型组合在非肝癌组的分布频率高于肝癌组(分别为P=0.001,OR=3.728和P=0.003,OR=4.032)。结论CTLA-4+49含A基因型和TIM-3-1516含T基因型可能是慢性HBV感染的危险因素,基因型组合增加了慢性HBV感染的发病风险。CTLA-4+49 GG基因型和TIM-3-1516 GT+TT基因型可能与HCC发生相关,基因型组合也增加了HCC发生的风险。CTLA-4和TIM-3基因多态性可能各自并联合地增加慢性HBV感染和肝细胞癌的风险。

Association of T regulatory cells with natural course and response to treatment with interferon-α in patients with chronic hepatitis B infection

Background Regulatory T cell populations, particularly CD4＋CD25＋ T regulatory cells, have been implicated in the persistence of hepatitis B virus （HBV） infection. However, no clear relationship has been established between the frequency of CD4~CD25~ T regulatory cells in the peripheral blood and either the disease phases in the natural history of chronic HBV infection or in the response to interferon-a therapy. Methods In the present study, three different common markers of CD4＋CD25＋ T regulatory cells were used to determine the numbers of T regulatory cells in healthy controls and in patients with chronic HBV infection. Results No significant difference was found when samples were gated for CD25hi and CD25＋FoxP3＋ T cells. A significant correlation was found between the number of CD4＋ Treg cells that gated with CD25＋FoxP3＋ and CD25＋CD127low/- in healthy controls and in patients with chronic hepatitis B （CHB） （r=0.67, 0.59; P 〈0.01）. The percentages of Treg cells were （8.56±2.01）% in asymptomatic carriers （Asc）, （8.74±3.04）% in inactive HBsAg carriers, （10.7±2.93）% in CHB and （7.42±1.28）% in healthy controls （F=-11.1, P 〈0.001）. The percentage of Treg cells in patients with CHB was higher than in asymptomatic HBV patients, inactive HBsAg carriers, or healthy controls （P 〈0.01）. The proportion of CD4＋CD25＋CD127 low/T cells in patients who responded to interferon-（] was （11.9±3.3）%, （9.1±2.4）% and （9.0±2.9）% at baseline, week 12 and week 24 after treatment, respectively （Z=2.42, P〈0.05; Z=2.67, P〈0.01）. Conclusions These results suggest that the proportion of the CD4＋CD25＋ regulatory T cells might be affected by the application of different markers in process to detect T regulatory cells. The frequency of Treg cells was increased in patients with CHB, which might be associated with the disease activity of these patients and contribute to prevention of extensive liver damage. A decline in Treg cells at week 12 of treatment might be associated with a better response to treatment with interferon-α.

Impact of baseline CD4^＋ T cell counts on the efficacy of nevirapinebased highly active antiretroviral therapy in Chinese HIV/AIDS patients： a prospective, multicentric study

Background CD4^＋T cell counts have been used as the indicator of human immunodeficiency virus type 1 （HIV-1） disease progression and thereby to determine when to start highly active antiretroviral therapy （HAART）. Whether and how the baseline CD4^＋T cell count affects the immunological and viral responses or adverse reactions to nevirapine （NVP）-containing HAART in Chinese HIV-1 infected adults remain to be characterized. Methods One hundred and ninety-eight HIV-seropositive antiretroviral therapy （ART）-naive subjects were enrolled into a prospective study from 2005 to 2007. Data were analyzed by groups based on baseline CD4^＋T cell counts either between 100-200 cells/μl or 201-350 cells/μl. Viral responses, immunologic responses and adverse events were monitored at baseline and at weeks 4, 12, 24, 36, 52, 68, 84, 100. Results Eighty-six and 112 subjects ranged their CD4^＋T cell counts 100-200 cells/μl and 201-350 cells/μl, respectively. The pre-HAART viral load in CD4 201-350 cells/μl group was significantly lower than that in CD4 100-200 cells/μl group （P=0.000）. After treatment, no significant differences were observed between these two groups either in the plasma viral load （pVL） or in the viral response rate calculated as the percentage of pVL less than 50 copies/ml or less than 400 copies/ml. The CD4^＋T cell counts were statistically higher in the 201-350 group during the entire follow-ups （P 〈0.01） though CD4^＋ T cell count increases were similar in these two groups. After 100-week treatment, the median of CD4^＋ T cell counts were increased to 331 cells/μl for CD4 100-200 cells/μl group and to 462 cells/μl for CD4 201-350 cells/μl group. Only a slightly higher incidence of nausea was observed in CD4 201-350 cells/μl group （P=0.05） among all adverse reactions, including rash and liver function abnormality. Conclusions The pVLs and viral response rates are unlikely to be associated with the baseline CD4^＋T cell counts. Initiating HAART in Chinese HIV-1 infected patients with higher baseline CD4^＋T cell counts could result in higher total CD4^＋T cell counts thereby achieve a better immune recovery. These results support current guidelines to start HAART at a threshold of 350 cells/μl.

Th17/Treg比例联合FIB-4和APRI评分对HBV相关肝纤维化的评估价值分析

目的研究辅助性T淋巴细胞17(Th17)/调节性T淋巴细胞(Treg)比例联合基于4因子的纤维化指数(FIB-4)、天门冬氨酸氨基转移酶/血小板比值指数(APRI)评分对乙型肝炎病毒(HBV)相关肝纤维化的评估价值。方法选择2019年6月-2021年6月内蒙古医科大学附属医院收治的160例HBV感染患者为研究对象,按照肝活检结果将其分为非肝纤维化组(S_(0)~S_(1))(n=58)和肝纤维化组(S_(2)~S_(4))(n=102)。比较两组Th17/Treg比例、FIB-4、APRI评分及其与肝纤维化的相关性,并分析三者单独及联合诊断肝纤维化的价值。结果与非肝纤维化组比较,肝纤维化组Th17/Treg比例[(1.24±0.31)vs.(1.61±0.43)]、FIB-4[(1.32±0.38)vs.(2.15±0.59)]、APRI[(0.64±0.20)vs.(1.51±0.48)]升高,差异均有统计学意义(P均<0.05)。经Spearman相关性分析显示,Th17/Treg比例、FIB-4、APRI均与肝纤维化病理分期成正相关(r=0.271、0.523、0.618,P均<0.05)。ROC曲线显示,Th17/Treg比例、FIB-4、APRI评分单独与联合评估HBV相关肝纤维化的敏感度分别为70.59%、82.35%、89.22%、92.16%,特异度分别为82.76%、89.66%、91.38%、96.55%,其中三者联合的评估价值最高。结论Th17/Treg比例联合FIB-4、APRI评分对HBV相关肝纤维化的评估价值较高,值得推广。

HBsAg致敏自体树突状细胞治疗乙肝病毒相关性慢加急性肝衰竭患者的临床效果

目的观察乙型肝炎(乙肝)表面抗原(HbsAg)致敏自体树突状细胞(抗HBV-DCs)治疗乙肝病毒相关性慢加急性肝衰竭(acute-on-chronic liver failure,ACLF)患者临床效应及对肿瘤坏死因子-α(TNF-α)、γ干扰素(IFN-γ)、白细胞介素-4(IL-4)水平的影响。方法选择住院治疗的64例ACLF患者为研究对象,抗HBV-DCs联合恩替卡韦(entecavir,ETV)治疗者为联合治疗组,ETV单独治疗者为单独治疗组,健康志愿者为对照组。观察治疗前后治疗组在外周血树突状细胞数量、频率、T淋巴细胞亚群及TNF-α、IFN-γ、IL-4水平的变化。结果两治疗组患者治疗前树突状细胞数量及频率较对照组水平低(P<0.05);联合治疗组与单独治疗组患者在治疗前树突状细胞数量及频率比较差异无统计学意义(P>0.05)。治疗后联合治疗组较单独治疗组上调明显(P<0.05);两治疗组在治疗前TNF-α、IFN-γ水平较对照组高,IL-4水平较对照组低,差异有统计学意义(P<0.05)。联合治疗组与单独治疗组在治疗前TNF-α、IFN-γ、IL-4水平间差异无统计学意义(P>0.05),治疗后联合治疗组TNF-α、IFN-γ水平下调较单独治疗组明显,联合治疗组IL-4水平上调水平较单独治疗组显著(P<0.05)。两治疗组治疗后TNF-α、IFN-γ水平下降明显,IL-4水平逐渐升高,治疗24周后复常,较治疗前比较差异有统计学意义(P<0.05)。治疗前两治疗组外周血CD3^+T淋巴细胞、CD4^+T淋巴细胞水平低于对照组,CD^8+T淋巴细胞水平高于对照组(P<0.05)。治疗前联合治疗组与单独治疗组间CD3^+T淋巴细胞、CD4^+T淋巴细胞、CD8^+T淋巴细胞水平差异无统计学意义(P>0.05)。治疗后联合治疗组CD3^+T淋巴细胞、CD4^+T淋巴细胞水平较单独治疗组上调明显(P<0.05)。ACLF患者治疗前外周血树突状细胞数量与CD3^+T淋巴细胞、CD4^+T淋巴细胞水平呈正相关(P<0.05),与CD8^+T淋巴细胞水平呈负相关(P>0.05)。结论抗HBV-DCs对调节树突状细胞数量及频率提升、激活T淋巴细胞、调节机体免疫失衡具有一定作用,可能成为HBV相关性肝病治疗的一种新路径。

慢性乙肝患者调节性T细胞的检测及临床价值

目的 通过检测慢性乙型肝炎患者外周血中CD4+CD25nt/hiCD127lo调节性T细胞比例以及替比夫定治疗前后调节性T细胞的变化,探讨慢性乙型肝炎的病理机制.方法 荧光标记的单克隆抗体染色外周血淋巴细胞,利用流式细胞仪检测CD4+CD25nt/hiCD127lo调节性T细胞比例.结果 慢性乙肝患者外周血CD4+CD25nt/hiCD127loTreg细胞占CD4+T细胞的比例为(7.5%±1.7%),较正常对照组显著增多(6.0%±1.4%,P【0.01),替比夫定治疗后则明显下降(6.5%±1.9%,P【0.01).结论 CD4+CD25nt/hiCD127loTreg细胞的增多与乙型肝炎的慢性化有关,替比夫定治疗可有效改善患者的免疫状态.

The Role of Immune Cells in Chronic HBV Infection

Hepatitis B virus(HBV)infection is a major cause of chronic liver diseases that may progress to liver cirrhosis and hepatocellular carcinoma.Host immune responses are important factors that determine whether HBV infection is cleared or persists.After infection,viral replication occurs inside hepatocytes,and the secretion of infectious virions can take place at high rates for decades.Consequently,HBV DNA and viral proteins,like HBV early antigen(HBeAg)and HBV surface antigen(HBsAg),can be easily detected in serum.Chronic infection with HBV is the result of an ineffective antiviral immune response towards the virus.In this review,we discuss the role of immune cells in chronic HBV infection.

Hepatic effector CD8＋ T-cell dynamics

CD8＋ T cells play a critical role in hepatitis B virus （HBV） pathogenesis. During acute, self-limited infections, these cells are instrumental to viral clearance; in chronic settings, they sustain repetitive cycles of hepatocellular necrosis that promote hepatocellular carcinoma development. Both CD8＋ T-cell defensive and destructive functions are mediated by antigen-experienced effector cells and depend on the ability of these cells to migrate to the liver, recognize hepatocellular antigens and perform effector functions. Understanding the signals that modulate the spatiotemporal dynamics of CD8＋ T cells in the liver, particularly in the context of antigen recognition, is therefore critical to gaining insight into the pathogenesis of acute and chronic HBV infection. Here, we highlight recent data on how effector CD8＋ T cells traffic within the liver, and we discuss the potential for novel imaging techniques to shed light on this important aspect of HBV pathogenesis.

Analysis of causes for liver function deteriora-tion in patients with HIV/HCV co-infection

BACKGROUND:Co-infection of hepatitis C virus (HCV) and human immunodeficiency virus type 1 ( HIV-1 ) is common in hemophiliacs and drug abusers. To assess the interaction between HIV and HCV disease progression, we examined 82 HIV/HCV co-infection patients and 62 HCV infection patients. METHODS: Liver function, pathological changes, infec- tion duration, immune function and qualitative HCV-RNA and HCV antibody were compared retrospectively between the two groups of patients. RESULTS: Fourty-eight patients (58.5%) in the HIV/ HCV co-infection group and 53 patients (85.5%) in the HCV infection group showed abnormal liver function. No significant difference was observed in inflammation and fi- brosis in the two groups P =0.187, 0.954). However, liver abnormality in the patients with HIV/HCV co-infection appeared 8 years earlier than in those with HCV infection alone (P<0.001). As to immune function, the counts of CD4+T and CD8+ T in the HIV/HCV group were (226.35 ± 173.49)×106/L and (914. 40 ±448. 28)×106/L, whereas in the HCV group they were (752.31±251.69)×l06/L and (529.011170.67)×106/L respectively. The difference in the two groups was highly significant (P<0.001; P<0.001). The ratio of the number of people with both HCV-RNA and HCV antibody positive to the number of HCV-RNA positive and HCV antibody negative in the HIV/HCV group was 52:9, whereas in the HCV group it was 44:1 (P = 0.043). CONCLUSION: HIV/HCV co-infection can accelerate de- terioration of hepatitis C, which may be due to the effect of HIV on cellular immunity and humoral immunity of the body.

慢性HBV感染者不同免疫状态下及抗病毒治疗后外周血T细胞亚群比例和细胞因子水平特点

目的：观察不同免疫状态的慢性乙型肝炎病毒（HBV）感染者及抗病毒治疗后外周血CD4^＋T、CD8^＋T细胞比例、CD4^＋/CD8^＋比值及血清中细胞因子IL-6、IL-10、TNF-α和IFN-γ水平的变化特点及与HBeAg血清学转换的相关性。方法纳入79例处于不同免疫状态的慢性HBV感染者（其中非活动性HBV携带期15例、免疫耐受期20例、免疫活化期44例）及21例健康对照者为研究对象，采用流式细胞术检测其外周血CD4^＋T、CD8^＋T细胞比例。对其中33例免疫活化期患者进行核苷（酸）类似物抗病毒治疗并随访至48周。分别于治疗前（T0期）及治疗后4周（T1期）、8周（T2期）、12周（T3期）、24周（T4期）和48周（T5期）采集外周血，流式细胞术检测各时间点患者外周血CD4^＋T、CD8^＋T细胞比例。同时对部分患者血清标本中的细胞因子水平进行检测。结果在慢性HBV感染者中，免疫活化组患者外周血CD4^＋T细胞比例及CD4^＋/CD8^＋比值显著低于健康对照组（P均＜0.05），而CD8^＋T细胞比例显著高于健康对照组（F =3.610，P ＜0.05）。抗病毒治疗后，△T0～T1、△T0～T2期，HBeAg血清学转换组外周血CD8^＋T细胞比例升高率显著高于未转换组（P均＜0.05）；△T0～T5期，HBeAg血清学转换组外周血CD4^＋T细胞比例升高率显著高于未转换组（Z =-2.200，P ＜0.05）。免疫活化组患者血清IL-10、IFN-γ水平均显著高于对照组和免疫耐受组（P均＜0.05）。免疫活化组患者抗病毒治疗后12周及24周，血清IL-10水平显著下降（t=3.037、3.180，P均＜0.05），与ALT水平、HBV DNA载量呈正相关关系（P均＜0.05）。结论慢性HBV感染者在不同免疫状态下外周血T细胞亚群比例及血清细胞因子水平表现不同，免疫活化期患者与健康对照者比CD4^＋T比例及CD4^＋/CD8^＋下降，CD8^＋T比例升高，血清IL-10水平显著升高。HBeAg阳性患者，核苷（酸）类似物抗病毒治疗过程中外周血CD8^＋T细胞和CD4^＋T细胞的比例升高可能有助于预测HBeAg的血清学转换。