HBV相关疾病患者血清IL-6、T淋巴细胞亚群、HBV DNA的变化及临床意义

目的探讨HBV相关疾病患者体内细胞因子、细胞免疫功能、病毒载量以及肝功能的变化。方法选取70例HBV相关疾病患者（慢性乙肝43例,肝硬化15例,慢性重型肝炎12例）和健康查体者18例（对照组）。应用ELISA法检测血清IL-6,流式细胞仪检测T淋巴细胞亚群,FQ-PCR检测HBV DNA,全自动生化仪检测肝功能。结果 HBV相关性疾病患者血清IL-6、ALT、CD4＋、CD8＋水平与对照组比较差异有统计学意义（P均〈0.05）,HBV DNA阳性组与HBV DNA阴性组比较I,L-6、CD4＋、CD8＋均有统计学差异（P均〈0.05）。结论 HBV相关疾病患者存在免疫功能紊乱I,L-6可作为判断病情、治疗效果和评估预后的重要指标。     

慢性乙型肝炎患者T细胞亚群，Mil-2R，sIL-2R，IL-6，IL-8，TNF-alpha变化及意义

目的探讨病毒性肝炎(乙肝)患者T细胞亚群,mIL-2R,sIL-2R,IL-6,IL-8,TNF-α与乙肝发病机制的关系.方法采用APAAP技术和ELISA法检测92例慢性乙型肝炎患者T细胞亚群,mIL-2R和血清sIL-2R,IL-6,IL-8,TNF-α水平.结果慢性乙型肝炎患者CD4+细胞数较正常低(P＜0.05),对照组和实验组(慢性乙肝轻度、慢性乙肝中重度、肝炎后肝硬变、重症肝炎)的CD4+数值分别为:(45.6±3.6)%,(39.7±10.2)%,(40.6±11.0)%,(39.0±6.5)%,(31.2±8.9)%.CD9+细胞数显著上升(P＜0.05或P＜0.01),对照组和实验各组依次为:(28.7±3.2)%,(35.4±9.5)%,(38.7±7.6)%,(42.2±9.4)%,以致CD4+/CD8+比值下降;mIL-2R显著低于正常对照组(P＜0.05),在PHA激活后与正常接近,但均较PHA激活前显著增高(P＜0.01);PHA激活前的mIL-2R对照组与实验组分别为:(15.69±4.32)%,(3.98±5.36)%,(9.37±5.48)%,(9.77±5.76)%,(9.58±5.45)%.PHA激活后mIL-2R对照组与实验组分别为:(69.82±5.36)%,(67.98±3.69)%,(69.11±2.19)%,(63.93±1.32)%,(66.32±5.26)%.慢性乙肝患者血清中sIL-2R,IL-6,IL-8,TNF-α分别为:(798.9±69.0)ng/L,(2806.2±211.7)ng/L,(480.6±32.4)ng/L.正常对照组＜100ng/L,且在慢性肝炎、肝硬变活动期与稳定期之间、重型肝炎的肝坏死与恢复期之间,血清IL-6,IL-8,TNF-α三项指标的差异有显著性,均P＜0.001.结论乙肝患者存在免疫调节紊乱,而免疫异常至少有部分原因是细胞因子的作用.     

慢性乙型肝炎患者T细胞亚群,mIL-2R,sIL-2R,IL-6,IL-8,TNF-α变化及意义

目的探讨病毒性肝炎(乙肝)患者T细胞亚群,mIL-2R,sIL-2R,IL-6,IL-8,TNF-α与乙肝发病机制的关系.方法采用APAAP技术和ELISA法检测92例慢性乙型肝炎患者T细胞亚群,mIL-2R和血清sIL-2R,IL-6,IL-8,TNF-α水平.结果慢性乙型肝炎患者CD4+细胞数较正常低(P＜0.05),对照组和实验组(慢性乙肝轻度、慢性乙肝中重度、肝炎后肝硬变、重症肝炎)的CD4+数值分别为:(45.6±3.6)%,(39.7±10.2)%,(40.6±11.0)%,(39.0±6.5)%,(31.2±8.9)%.CD9+细胞数显著上升(P＜0.05或P＜0.01),对照组和实验各组依次为:(28.7±3.2)%,(35.4±9.5)%,(38.7±7.6)%,(42.2±9.4)%,以致CD4+/CD8+比值下降;mIL-2R显著低于正常对照组(P＜0.05),在PHA激活后与正常接近,但均较PHA激活前显著增高(P＜0.01);PHA激活前的mIL-2R对照组与实验组分别为:(15.69±4.32)%,(3.98±5.36)%,(9.37±5.48)%,(9.77±5.76)%,(9.58±5.45)%.PHA激活后mIL-2R对照组与实验组分别为:(69.82±5.36)%,(67.98±3.69)%,(69.11±2.19)%,(63.93±1.32)%,(66.32±5.26)%.慢性乙肝患者血清中sIL-2R,IL-6,IL-8,TNF-α分别为:(798.9±69.0)ng/L,(2806.2±211.7)ng/L,(480.6±32.4)ng/L.正常对照组＜100ng/L,且在慢性肝炎、肝硬变活动期与稳定期之间、重型肝炎的肝坏死与恢复期之间,血清IL-6,IL-8,TNF-α三项指标的差异有显著性,均P＜0.001.结论乙肝患者存在免疫调节紊乱,而免疫异常至少有部分原因是细胞因子的作用.     

毛细支气管炎患儿外周血T细胞亚群、IL-2及IL-6的检测及意义

目的探讨毛细支气管炎患儿外周血T细胞亚群、IL-2及IL-6的变化及意义。方法选择73例急性期呼吸道合胞病毒（RSV）毛细支气管炎患儿（毛细支气管炎组）和40例健康体检儿童（健康对照组）,采用免疫荧光法检测两组外周血T细胞亚群,ELISA法测定血清IL-2和IL-6。结果与健康对照组相比,毛细支气管炎组CD4＋水平和CD4＋/CD8＋比值增高,血清IL-2水平和IL-2/IL-6比值降低,IL-6水平增高。结论婴幼儿RSV毛细支气管炎与哮喘时T细胞亚群和Th1/Th2功能失调的表现类似。     

乙型肝炎患者外周血T细胞亚群和可溶性IL_2R的研究

目的探讨急慢性乙型肝炎患者外周血Ｔ细胞亚群及可溶性ＩＬ＿２Ｒ水平的变化，为诊断病情演变和预后判定寻找可靠的依据．方法应用ＡＰＡＡＰ桥联酶标法和双抗体夹心ＥＬＩＳＡ法检测正常人３０例和各型乙型肝炎患者１７５例的Ｔ细胞亚群变化和血清可溶性ＩＬ＿２Ｒ（ｓＩＬ＿２Ｒ）水平．结果各组乙肝患者ＣＤ３，ＣＤ４，ＣＤ４／ＣＤ８比值明显降低，ＣＤ８则明显升高．乙肝患者血清ｓＩＬ＿２Ｒ水平均显著升高，其中慢重肝、肝硬变、急性乙肝和慢性肝炎增高最为显著，与正常对照组相比Ｐ均＜００１，且与ＨＢＶ复制有关．结论乙型肝炎患者存在Ｔ细胞比例失衡，血清ｓＩＬ＿２Ｒ水平均明显增高，其水平与机体免疫功能状态及肝细胞损伤程度相关．     

老年肺癌患者IL-6、sIL-2R和T细胞亚群检测的临床意义

目的　探讨老年肺癌患者外周血 IL - 6、s IL- 2 R和 T细胞亚群活性的变化。方法　对老年肺癌患者化疗前、后及老年健康对照组应用流式细胞分析法测定 T细胞亚群 ,双抗体夹心法 (EL ISA)测定 IL - 6及 s IL- 2 R水平。结果　肺癌组 s IL - 2 R、IL- 6及 CD8高于对照组 (P<0 .0 5) ,而CD3 及 CD4水平低于对照组 ,化疗后 s IL - 2 R、IL- 6及 CD8水平明显下降 ,但仍高于对照组 ,CD3 及 CD4水平略升高 ,仍低于对照组。结论　 IL - 6、s IL-2 R及 T细胞亚群联合测定可以反映肺癌病人机体免疫功能状态 ,可作为判断疾病的预后和免疫调节治疗的客观指标。     

小儿重型肝炎患者外周血T淋巴细胞亚群的变化与临床意义

目的了解小儿与成人重型肝炎外周血CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量变化的差别,探讨小儿重型肝炎患者外周血T淋巴细胞亚群数量变化与临床意义。方法对35例小儿及30例成人重型肝炎进行外周血CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量检测。结果小儿重型肝炎成活的患儿外周血CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量明显高于死亡组（P〈0．01）：小儿重型肝炎中1岁以内患儿外周血CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量明显高于2～6岁组（P〈0．01）;小儿重型肝炎组CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量明显高于成人重型肝炎组（P〈0．01）;1岁以内的患儿病死率最高（P〈0．05）。结论／JxJL与成人重型肝炎外周血CD3^＋、CD3^＋CD4^＋及CD3^＋CD8^＋T淋巴细胞数量有明显差异,年龄越小,CD3^＋、CD3^＋CD4＋及CD3^＋CD8^＋T淋巴细胞数量越高;dxJL细胞免疫与成人明显不同,年龄越小,病情越重;病死率越高。     

乙型肝炎患者外周血T细胞亚群的分析及意义

目的研究乙肝患者外周血T细胞亚群的变化,进而了解其细胞免疫状态水平。方法用流式细胞仪对228例不同乙肝患者的外周血T细胞亚群CD+3、CD+4、CD+8绝对细胞计数及CD+4/CD+8比值进行检测,并将检测结果与e抗原、HBV-DNA含量、YMDD变异情况进行比较,以探讨各组之间的关系。结果与正常对照组相比,急性乙型肝炎(AHB)患者外周血T细胞亚群CD+3、CD+4、CD+8绝对细胞数升高(P<0.01);而慢性乙型肝炎(CHB)、肝硬化(LC)、原发性肝癌(PHC)患者依次降低,以LC、PHC两组降低最为显著(P<0.01);CD+4/CD+8比值AHB、CHB、LC、PHC 4组有依次升高的趋势。同时e抗原(-)、HBV-DNA(-)、YMDD变异株乙肝患者外周血T细胞亚群绝对细胞数明显低于e抗原(-)、HBV-DNA(-)、YMDD野生株乙肝患者(P<0.01或P<0.05);而CD+4/CD+8比值均高于对照组。结论不同乙肝患者体内存在T细胞亚群失衡及细胞免疫功能紊乱。     

慢性HBV感染及其相关肝病患者外周血T细胞亚群的变化

目的观察不同临床类型HBV感染者外周血T淋巴细胞亚群的差异,探讨HBV对人体T细胞免疫的影响及其可能机制,不同类型慢性HBV感染者免疫失衡的规律。方法用流式细胞仪技术检测患者外周血T细胞亚群。慢性HBV感染者318例,其中HBV携带者8例,慢性乙型肝炎231例,肝炎后肝硬化61例,原发性肝癌18例,观察患者的T淋巴细胞亚群、HBV DNA等。同时收集22名健康志愿者的新鲜血检测T淋巴细胞亚群。结果不同临床类型HBV感染者外周血CD3＋T、CD4＋T、CD8＋T细胞百分数低于正常对照组,差别均有统计学意义（P〈0.05或P〈0.01）,CD4＋/CD8＋比正常对照组低,但差异均无统计学意义（P〉0.05）。结论慢性HBV感染随着病情进展,由慢性肝炎→肝硬化→肝癌,直至细胞免疫功能逐渐衰退。慢性HBV感染的不同阶段的细胞免疫紊乱各具特点,就不同类型患者应采用不同的免疫调节治疗手段。     

慢性乙型肝炎患者不同免疫状态下外周血自然杀伤细胞亚群的特点及其临床意义

背景:外周血自然杀伤细胞(NK细胞)亚群在各种病毒感染性疾病中有不同程度的改变,其在HBV感染不同免疫状态下的变化尚不明确。目的:研究慢性乙型肝炎(CHB)患者不同免疫状态下外周血NK细胞亚群的变化特点及其临床意义。方法:采用流式细胞术检测46例CHB患者(免疫耐受期25例,免疫清除期21例)和10例健康志愿者的外周血NK细胞亚群比例,采用实时荧光定量PCR技术检测血清HBV DNA载量,同时检测血清ALT水平。结果:免疫清除期CHB患者外周血CD56~(bright)NK细胞比例明显高于免疫耐受期和健康对照组,差异均有统计学意义(0.877±0.493对0.647±0.294和0.546±0.173,P0.05);三组间CD56~(dim)NK细胞比例差异无统计学意义。外周血CD56~(bright)NK细胞比例与血清HBV DNA载量在免疫耐受期呈负相关(r=-0.575,P0.05),在免疫清除期则无相关性。各组不同免疫状态下外周血NK细胞亚群比例与血清ALT水平均无相关性。结论:外周血CD56~(bright)NK细胞比例增高可能是反映CHB患者进入免疫清除期的潜在指标之一,NK细胞及其亚群在抗HBV的适应性免疫中发挥重要调节作用。     

炎症性肠病患者T淋巴细胞亚群的改变及临床意义

目的探讨炎症性肠病(IBD)患者外周血T淋巴细胞亚群的变化特点及临床意义.方法对临床确诊的18例溃疡性结肠炎(UC)和9例Crohn病(CD)患者,于疾病的不同时期用荧光单克隆抗体标记-流式细胞仪技术检测其外周血T淋巴细胞亚群的变化,并结合临床和病理资料进行综合分析.结果UC患者缓解期时的CD3+、CD8+和自然杀伤(NK)细胞与对照组比较无显著差异;但活动期时,其CD8+和NK细胞数明显下降(CD8+活动期23.1%±1.0%,对照组31.3%±1.2%,P＜0.05;NK活动期9.8%±1.0%,对照组15.1%±1.1%,P＜0.01),CD4+/CDs+比值上升(活动期1.8±0.1,对照组1.4±0.02,P＜0.05);活动期CD患者的CD8+细胞和CD4+/CD8+比值(43.0%±44%和0.6±0.1)变化却与UC组活动期呈相反趋势(P＜0.01).结论T淋巴细胞亚群改变在IBD的发病机制中可能起重要作用;UC和CD的细胞免疫机制不同;外周血CD4+、CD8+、NK细胞和CD4+/CD8+比值可作为检测UC病情变化和疗效考核的敏感指标.     

miR-197 Expression in Peripheral Blood Mononuclear Cells from Hepatitis B Virus-Infected Patients

Background/Aims

This study aimed to investigate the microRNA (miRNA) expression profiles in peripheral blood mononuclear cell (PBMC) of hepatitis B virus (HBV)-infected patients with different clinical manifestations and to analyze the function of miR-197.

Methods

PBMC miRNA expression profiles in 51 healthy controls, 70 chronic asymptomatic carriers, 107 chronic hepatitis B patients, and 76 HBV-related acute on chronic liver failure patients were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). miR-197 mimic and inhibitor were transfected in THP-1 cells. qRT-PCR and ELISA for interleukin (IL)-18 mRNA and protein levels were performed, respectively.

Results

The microarray analysis revealed that 17 PBMC miRNA expression profiles (12 miRNAs downregulated and five miRNAs upregulated) differed significantly in HBV-induced liver disease patients presenting with various symptoms. The qRT-PCR results suggested that the PBMC miR-197 levels regularly decreased as the severity of liver disease symptoms became aggravated. IL-18, a key regulator in inflammation and immunity, was inversely correlated with miR-197 levels. Bioinformatic analysis indicated that IL-18 was a target of miR-197. Exogenous expression of miR-197 could significantly repress IL-18 expression at both the mRNA and protein levels in THP-1 cells.

Conclusions

We concluded that multiple PBMC miRNAs had differential expression profiles during HBV infection and that miR-197 may play an important role in the reactivation of liver inflammation by targeting IL-18.     

Hepatitis B Splice-Generated Protein Antibodies in Syrian Chronic Hepatitis B Patients: Incidence and Significance

Background:

Previous studies have suggested hepatitis B splice-generated protein (HBSP), when expressed, is involved in the pathogenesis of HBV infection.

Objectives:

We aimed to evaluate anti-HBSP incidence and association with several HBV infection parameters in a group of Syrian chronic hepatitis B patients.

Patients and Methods:

Eighty treatment-naïve HBsAg-positive adult chronic hepatitis B patients'' sera were included in our prospective targeted study. Liver function, virological and histological tests results were obtained from patients’ medical files. Three variants of a 20-mer HBSP-derived peptide were designed based on HBV genome sequences obtained from Syrian patients'' sera (GenBank Accession No. {"type":"entrez-nucleotide-range","attrs":{"text":"JN257148-JN257217","start_term":"JN257148","end_term":"JN257217","start_term_id":"364505024","end_term_id":"364505559"}}JN257148-JN257217). Microtiter plate wells were coated with the synthetic peptides and used to detect anti-HBSP antibodies by an optimized indirect enzyme-linked immunosorbent assay (ELISA). Samples were considered positive when showed optical density (OD) values higher than the cut-off value for at least one peptide variant.

Results:

Seven out of eighty (9%) CHB patients were positive for anti-HBSP antibodies. Mean OD values were not significantly different between HBeAg-positive and -negative patients (P > 0.05). OD values showed weak positive correlation with ALT and AST values (P < 0.05), and weak to moderate positive correlation with liver biopsy staging ranks (P < 0.05). No significant correlation was revealed with viral load values or liver biopsy grading ranks (P > 0.05).

Conclusions:

We introduced an anti-HBSP antibodies ELISA, designed for locally circulating HBV strains. Correlation observed of Anti-HBSP with liver fibrosis staging regardless of viral replication and liver inflammation suggests anti-HBSP antibodies as possible indicator for HBV-associated liver fibrosis.     

慢性乙型病毒性肝炎患者血浆内毒素、白介素-6和白介素-8水平的研究

目的探讨血浆内毒素、IL-6、IL-8水平与慢性肝病炎症活动、肝细胞损害的关系,揭示内毒素、IL-6、IL-8在慢性乙型肝炎发病机制中的作用。方法采用鲎试剂与合成基质(鲎三肽)的偶氮显色法和ELISA法分别检测患者血浆内毒素、IL-6、IL-8。结果慢性肝病组血浆内毒素、IL-6、IL-8水平均高于正常对照组(P<0.001);相关性分析三者间呈正相关;慢性肝病组按血清丙氨酸氨基转移酶(ALT)、血清门冬氨酸氨基转移酶(AST)高低分组后,血浆内毒素、IL-6、IL-8含量由高到低依次为>200u/L、100～200u/L、<100u/L组。结论血浆内毒素、IL-6、IL-8可导致肝细胞的损害,参与慢性乙型肝炎的发病,且三者密切相关。内毒素、IL-6、IL-8的高低在一定程度上反映了肝细胞的损害程度,为临床估计病情,判断预后,制定治疗方案提供参考指标。     

乙型肝炎患者外周血单个核细胞IL-6 mRNA 表达的定量分析

目的 探讨IL-6参与乙型肝炎的免疫病理操作和病毒清除的机理。方法 采用ELISA法检定了慢性乙型肝炎（chronic hepatitis B.CHB）患者血清IL-6含量,竞争性RT-PCR-ELISA法分析了CHB外周血单个核细胞的IL-6 mRNA表达水平。结果 CHB血清IL-6含量明显升高,竞争性RT-PCR结果表明对照组和患者组IL-6mRNA靶片段表达频率分别为41.7%（5/12）和82.1(23/28）,定量ELISA表明5例对照组IL-6 mRNA拷贝数在1000-10000之间,平均拷贝数的对数为3.65;而患者组均大于10000,平均拷贝数的对数为4.40。 结论 IL-6在乙型肝炎发病中具有重要作用,而IL-6水平升高主要与患者外周血单个核细胞（peripheral blood monocyte cell,PBMC)IL-6 mRNA表达增加有关。     

慢性乙型肝炎患者HBV DNA载量与血清IL-2、IL-6、IL-8、IL-13水平的相关性研究

目的探讨慢性乙型肝炎病毒(HBV)感染患者血浆HBV DNA载量与乙肝血清标志物的关系及与白细胞介素2(IL-2)、白细胞介素6(IL-6)、白细胞介素8(IL-8)、白细胞介素13(IL-13)含量变化的相关性。方法选取60例慢性HBV感染者(疾病组),采用免疫发光法定量检测乙型肝炎标志物;实时荧光定量PCR(FQ-PCR)检测HBV DNA载量,并以5.0×102copies/ml为临界点分为高载量组、低载量组;酶联免疫吸附试验(ELISA)检测IL-2、IL-6、IL-8、IL-13含量;并与20名健康对照者(正常对照组)比较。结果疾病组血浆HBV DNA载量5.0×102copies/ml时,以HBs Ag、抗-HBe、抗-HBc 3项指标升高为主要表现,阳性检出率为95%,IL-2、IL-6、IL-8、IL-13水平显著高于正常对照组(P0.05);血浆HBV DNA5.0×102copies/ml时,以HBs Ag、HBe Ag、抗-HBc 3项指标升高为主要表现,疾病组阳性检出率为70%,抗-HBe阳性检出率为30%;疾病组血清IL-2、IL-6、IL-8、IL-13水平显著高于对照组(P0.05),并显著高于血浆HBV DNA低载量组(P0.05)。结论 HBV DNA载量和细胞因子水平的变化可反应机体免疫活动状况,对临床观察疗效具有重要意义。     

Prevalence of Occult Hepatitis B Virus in Plasma and Peripheral Blood Mononuclear Cell Compartments of Patients With Chronic Hepatitis C Infection in Tehran-Iran

Background

Occult hepatitis B virus (HBV) infection (OBI) is frequently reported in patients with chronic hepatitis C virus (HCV) infection. An association between OBI and more liver damage, cirrhosis, hepatocellular carcinoma, and reduced response to interferon therapy in patients with HCV infection is suggested.

Objectives

The aim of this study was to determine the prevalence of occult HBV, and evaluate its clinical influence on patients with chronic HCV.

Patients and Methods

A cohort study including50 patients with positive results for HCV, and negative results for HBsAg tests was performed. The patients were divided into two groups: one group had positive results for both HCV and occult HBV tests (n = 18), and the other had positive results for HCV, but negative findings for occult HBV (n = 32). All were treated with PEG-IFN alpha-2a and Ribavirin. Presence of HCV RNA was followed in these patients.

Results

HBV-DNA was detected using nested-PCR in 20% of plasma and 32.6% of peripheral blood mononuclear cell (PBMC) compartments. No significant differences were observed between patients with and without occult HBV for sex, age, duration of HCV infection, histological markers, presence of anti-HBc, HCV viral load, and HCV genotype. The response rate was significantly higher in patients with positive results for HBV-DNA test compared to those with negative findings (100% vs. 71.9 %, P < 0.05).

Conclusions

In conclusion, occult HBV was found in 36% of patients with negative results for HBsAg, but positive results for HCV. Detection of HBV-DNA in both PBMCs and plasma together in comparison with plasma alone provided more true identification of OBI.The SVR rate was significantly higher in coinfected patients than mono-infected ones.     

慢性乙型肝炎患者外周血树突细胞的体外扩增及鉴定

目的尝试从慢性乙型肝炎患者外周血扩增培养出大量树突细胞,为进一步临床应用树突细胞治疗慢性乙型肝炎提供物质基础.方法分别取健康人和患者外周血分离培养单核细胞,加入rhGM-CSF、rhIL-4,经7天培养后收获即为不完全成熟树突细胞(DC).以倒置显微镜观察摄影、透射电镜摄影、流式细胞仪测定表面特异性分子的表达、混合淋巴细胞反应测定功能,对DC进行鉴定.结果第七天记数有明显树突状突起的大个细胞或细胞团,产率为0.2～4.5×105/6ml血;透射电镜可见细胞表面伸展的突起;70%以上细胞有DC特异性标志CD1a高表达,而单核细胞特异性标志CD14表达小于4%;能刺激同种异体淋巴细胞强烈增殖,随刺激细胞数目的增加,其促增殖作用增强,P<0.01.结论在本实验条件下,从慢性乙型肝炎患者外周血单核细胞可诱导扩增出大量DC.