慢性 HBV 感染者自发清除 HBsAg 后免疫功能的观察

目的：了解慢性 HBV 感染者自发清除 HBsAg 后,外周血细胞免疫和体液免疫功能变化。方法流式检测67例慢性 HBV 感染者自发清除 HBsAg 后外周全血细胞亚群,包括：T 淋巴细胞亚群比值、B 淋巴细胞计数比值和 NK 细胞计数比值。同时检测血清 IgG、IgA 及 IgM 和 ALT、AST。以23例 HBeAg 阳性慢性 HBV 感染者、323例 HBeAg 阴性慢性 HBV 感染者和43例乙肝血清学标志物全阴健康人（排除其他疾患）作对照。结果HBsAg 自发清除组,总 T 淋巴细胞计数比值为69．05％±8．32％,CD4＋ T 淋巴细胞计数比值为38．95％±6．93％, CD8＋ T 淋巴细胞计数比值为26．97％±7．86％,CD4＋/CD8＋比值为1．61％±0．70％,B 淋巴细胞计数比值为12．27％±4．23％和 NK 细胞计数比值为18．15％±8．67％。HBsAg 自发清除组 CD4＋与 CD4＋/CD8＋比值均高于其余各组（P ＜0．05）,而总 T 淋巴细胞计数比值、CD8＋ T 淋巴细胞计数比值、B 淋巴细胞计数比值和 NK 细胞计数比值在各组间差异不明显（P ＞0．05）。HBeAg 阳性 HBV 感染组与健康人对照组比较,CD4＋ T 淋巴细胞计数比值明显降低（P ＝0．034＜0．05）,而 CD4＋/CD8＋比值降低差异无统计学意义（P ＞0．05）。HBeAg 阴性 HBV 感染组细胞免疫功能与健康人对照组比较差异无统计学意义（P ＞0．05）。四组人群血清免疫球蛋白检测结果组间比较差异无统计学意义（P ＞0．05）。结论慢性 HBV 感染者自发清除 HBsAg 后,CD4＋ T 淋巴细胞计数比值和 CD4＋/CD8＋比值明显增高,说明 CD4＋ T 淋巴细胞可能是 HBsAg 自发清除过程中最为关键的因素。     

流式细胞仪测定肝病患者T淋巴细胞亚群及NK细胞

目的：研究慢性乙型肝炎、肝硬化及原发性肝癌（PHC）患者的免疫状态,观察T细胞亚群以及自然杀伤（NK）细胞活性在不同肝病中的变化情况。方法：应用流式细胞仪测定了共147例肝病患者血中T细胞亚群和NK细胞活性。结果：与正常组相比较,各病例组CD3＋及CD4＋细胞数减少,CD8＋细胞数则高（P＜0．05）,使CD4＋／CD8＋比值明显降低（P＜0．01）,其中,肝硬化上PHC组变化更为明显,PHC组的上述指标甚至与慢性乙直炎组之间也存在着差异（P＜0．0）。同时PHC组NK细胞活性明显低于正常对照组,甚至低于慢性乙型肝炎和肝硬化组（P＜0．05）,但其他三组间无显著差别（P＞0．05）,结论：用流式细胞仪检测肝病虱外周血中T淋巴细胞亚群及NK细胞活性具有快速、灵敏、准确等特点,其检验结果,对了解肝病患者的免疫状态态,指导肝病的治疗及判断预后具有重要价值。     

消化系肿瘤T细胞亚群NK B细胞表面抗原检测的意义

目的 了解消化系统肿瘤患者的免疫状态,探讨T细胞亚群、NK细胞、B细胞表面抗原与消化系统肿瘤的关系。方法 采用流式细胞仪分别检测34例消化系统肿瘤患者外周血T细胞亚群：CD3＋、CD4＋、CD8＋,以及CD19＋、CD16＋CD56＋的淋巴细胞百分数,其中食道癌10例,胃癌8例,肝癌6例,大肠癌10例,并与20例政党对照比较。结果 消化系统肿瘤患者CD3＋、CD4＋、CD16＋CD56＋均较正常对照组低（P＜0．05）,CD8＋高于对照组（P＜0．05）,CD19＋与正常对照无显著差异（P＞0．05）,各免疫指标在消化系统肿瘤之间无显著差异（P＞0．05）。结论 消化系统肿瘤患者细胞免疫功能普遍低下,T细胞亚群、NK细胞、B细胞表面抗原检测可作为消化系统肿瘤患者免疫功能的监测指标。     

成人原发性肾病综合征T细胞亚群、NK细胞及B细胞的变化及临床意义

目的探讨成人原发性肾病综合征（primarynephrotiesyndrome,PNS）患者T淋巴细胞亚群、NK细胞及B淋巴细胞的变化及临床意义。方法应用流式细胞仪检测64例成人PNS活动期患者外周血中T淋巴细胞亚群（CD3＋、CD3＋CD4＋、CD3＋CD8＋、CD4＋／CD8＋）、CD16＋CD56＋NK细胞及CD19＋B淋巴细胞水平的变化,同时检测30例同期来院体检的正常人作为对照。结果PNS组与对照组相比：CD3＋、CD3＋CD4＋T淋巴细胞降低．CD3＋CD8＋T淋巴细胞升高,CD4＋／CD8＋比值显著下降且差异均有统计学意义（P均〈0．05）;PNS组CD16＋CD56＋NK细胞较对照组降低,而CD19＋B淋巴细胞显著升高,且差异亦均具有统计学意义（P均〈0．05）。结论成人PNS活动期T细胞亚群的比例结构发生严重失衡,细胞免疫功能降低,而体液免疫功能亢进。     

乙型肝炎病毒感染者在有或无肝损情况下的T细胞亚群情况分析

目的：观察乙型肝炎病毒（HBV）感染者不同肝损情况下的 T 细胞亚群的分布情况,探讨 HBV 携带者肝脏损伤与免疫功能紊乱的关系。方法用实时荧光定量（RT-PCR）方法检测 HBV-DNA,用流失细胞仪直接免疫荧光法检测外周血 T 细胞亚群;用 A2400全自动生化分析仪检测丙氨酸氨基转移酶（AST）。结果 HBV-DNA 阳性且 ALT 正常者与健康体检者比较,CD3＋ T 细胞、CD4＋ T 细胞和 CD4/CD8均下降,CD8＋ T 细胞升高,差异有统计学意义（P ＜0．05）;HBV-DNA 阳性且 ALT 异常者与健康体检者比较,CD3＋ T 细胞、CD4＋ T 细胞和CD4/CD8均下降,CD8＋ T 细胞升高,差异有统计学意义（P ＜0．05）;HBV-DNA 阳性且 ALT 正常者与 HBV-DNA阳性且 ALT 异常者比较,CD3＋ T 细胞和 CD4＋ T 细胞均有下降趋势,但差异无统计学意义（P ＞0．05）;CD8＋ T 细胞显著升高,CD4/CD8显著降低,差异有统计学意义（P ＜0．05）。结论HBV 感染者在有或无肝损情况下,T 淋巴细胞亚群会发生不同程度的紊乱,连续监测 HBV 携带者的 T 淋巴细胞亚群能为临床诊断和治疗提供依据。     

外周血 DNT 细胞及 T细胞亚群变化与乙型肝炎慢性化的相关性研究

目的：探讨外周血中CD3＋CD4－CD8－（ DNT）细胞及T细胞亚群变化与乙型肝炎病毒（ HBV）感染慢性化的关系。方法用流式细胞术测定136例HBV感染者[33例无症状携带者（ ASC ）、28例急性乙型肝炎（AHB）、28例轻度慢性乙型肝炎（CHB）、25例中度CHB、22例重度CHB患者]及39名健康对照（HC）外周血中的DNT细胞及T细胞亚群。结果 ASC组、AHB组、HC组DNT细胞比例分别为5．43％±3．31％、4．75％±2．71％和4．82％±3．43％,3组间差异无统计学意义（P＞0．05）,其中HC组和AHB组均低于CHB各组（由轻到重分别为7．97％±4．12％、8．41％±4．93％、11．36％±5．01％,P均＜0．05）,但ASC组仅显著低于重度CHB组（P＜0．05）,与轻度CHB组和中度CHB组间差异均无统计学意义（P＞0．05）;DNT细胞比例在轻度CHB组与中度CHB组间差异无统计学意义（P＞0．05）,但二组均显著低于重度CHB组（P均＜0．05）。 HC组、AHB组、ASC组间T淋巴细胞亚群分布差异无统计学意义（P均＞0．05）,但随着CHB患者病情加重CD3＋、CD3＋CD4＋CD8－（ CD4＋）细胞比例降低,CD3＋CD4－CD8＋（ CD8＋）细胞比例升高。结论外周血DNT细胞比例的升高与HBV感染者慢性化关系密切,且与CHB患者的疾病进程有关。     

妊娠合并HBV感染者外周血淋巴细胞亚群和NK细胞的改变

目的探讨孕妇妊娠早、中、晚期合并HBV感染者外周血淋巴细胞哑群和NK细胞表达水平的变化。方法流式细胞术检测孕妇怀孕10周、14周和27周HBV感染者外周血CD4^＋．CD8^＋T细胞和NK细胞的表达水平。结果与正常孕期比较，怀孕中期合并HBV感染者其CD4^＋和CD8T细胞表达水平下降（t=29．451。t=16．183．均P〈0．05）。孕晚期合并HBV感染者除上述指标外，CD4^＋/CD8^＋比例和NK细胞表达水平也下降（t=18．962，t=16．372．t=7．662，t=7．662，t=26．696，均P〈0．05）。而孕早期合并HBV感染者组上述指标无明显差异（P〉0．05）。早产2例（2／40），无死胎。结论妇女孕期合并HBV感染者。孕早期外周血淋巴细胞亚群和NK细胞表达水平无明显变化．而怀孕中期和晚期外周血淋巴细胞亚群和NK细胞表达水平有明显变化。     

系统性红斑狼疮患者外周血淋巴细胞亚群分析

目的探讨T淋巴细胞亚群、B淋巴细胞和NK细胞在系统性红斑狼疮fSLE忡的作用。方法利用流式细胞仪对29例活动期、24例非活动期SLE患者及50例健康对照者的外周血中的CD3＋、CD4＋、CD8＋、CD3-CD16＋CD56＋（NK）及CD19＋（B）淋巴细胞进行检测。结果活动期组与非活动期组及健康对照组比较：CD4＋T淋巴细胞、CD4＋／CD8＋比值及NK细胞明显降低（P〈0．05）,B淋巴细胞明显增高（P〈0．05）,CD3＋T和CD8＋T淋巴细胞无统计学意义（P〉0．05）。非活动期组与健康对照组比较．T淋巴细胞各亚群、NK细胞及B淋巴细胞均无统计学意义（P〉0．05）。结论检测外周血淋巴细胞亚群．对判断病情及指导临床治疗具有重要意义。     

免疫治疗联合化疗对多发性骨髓瘤患者外周血T、B淋巴细胞及调节性T细胞水平的影响

【目的】探讨免疫联合化疗对多发性骨髓瘤（MM）患者外周血T、B淋巴细胞及调节性T细胞（Tregs）水平的影响。【方法】本院收治的MM患者60例,采用数字随机对照表分为两组,每组各30例;对照组给予DC-CIK免疫治疗＋化疗,对照组仅给予化疗,评估两组临床疗效,观察两组T淋巴细胞、Tregs水平、B细胞、浆细胞比例水平变化;随访1年,比较两组复发率。【结果】观察组治疗有效率为53．33％（i6／30）显著高于对照组26．67％（8／30）,1年复发率为6．67％（2／30）显著低于对照组30．0％（9／30）,且差异有显著性（P〈0．05）。观察组治疗后CD4＋T淋巴细胞比例、CD4＋／CD8＋T淋巴比值显著高于对照组,CD8＋T淋巴细胞比例显著低于对照组,且差异有显著性（P〈0．05）。观察组治疗后CD4＋CD25＋Tregs、CD4＋CD25 high Tregs显著低于对照组,且差异有显著性（P〈0．05）。观察组治疗后B细胞比例高于对照组,浆细胞比例低于对照组,且差异有显著性（P〈0．05）。【结论】DCcIK免疫治疗＋化疗能够改善MM患者免疫功能紊乱症状,降低浆细胞比例,提高临床疗效。     

慢性HBV感染者的免疫状态与外周血T、B细胞亚群的含量相关性研究

目的通过分析慢性HBV感染者不同免疫状态外周血T、B细胞亚群的占淋巴细胞的百分比探讨其相互关系。方法选取2010年2月至2013年3月50例在该院就诊医治的慢性HBV感染者作为试验组,按照免疫状态的不同将试验组分为21例免疫耐受组和29例免疫清除组,同时选取20例健康成人作为正常对照组,采用流式细胞术检测外周血T、B细胞亚群的百分比,对3组的临床资料进行了对比。结果与对照组相比,免疫耐受组和免疫清除组患者的B细胞、CD8+T细胞的百分比明显增高;而免疫耐受组和免疫清除组患者CD3+T、CD4+T细胞的百分比和CD4+/CD8+的比值均明显降低,不同组间相比较差异有统计学意义(P0.05)。结论外周血T、B细胞亚群的百分比可作为慢性HBV感染者免疫状态的参考指标,常规检测该项目对了解HBV感染者的不同免疫状态以及疾病的治疗和疫苗接种等具有重要的指导作用,值得临床推广应用。     

Hepatitis B virus DNA and hepatitis B surface antigen levels in chronic hepatitis B

Despite universal vaccination, chronic hepatitis B (CHB) continues to be a major health burden worldwide, with an estimated 350–400 million people infected with the virus. Over the past decade, rapid progress has been made with regards to antiviral therapy for CHB, from conventional interferon to pegylated interferon, and with the earliest oral agent lamivudine to the current, more potent drugs such as entecavir and tenofovir. There have also been new developments in the diagnostic and monitoring tools for CHB. Qualitative hepatitis B surface antigen (HBsAg) testing has been used to diagnose patients infected with CHB. More recently, quantitative HBsAg titers have been used to predict treatment outcome when measured at baseline or early into treatment. The progress on the use of hepatitis B virus (HBV) DNA levels has been more rapid. Serum HBV DNA levels have been shown to be important in the natural history of CHB infection, with higher levels being significantly associated with the development of cirrhosis and hepatocellular carcinoma. For patients receiving antiviral therapy, the baseline and early on-treatment HBV DNA levels are important in determining treatment outcomes. Monitoring of HBV DNA levels during therapy will allow for early detection of drug resistance. The end-of-treatment and post-treatment HBV DNA levels have been demonstrated to be important indicators of treatment success and relapse, respectively. With newer and more powerful antiviral agents, and with the development of quantitative assays that are highly sensitive, further studies are needed to optimize the use of these tools and agents in the modern management of CHB.     

Hepatitis B genotypes in chronic hepatitis B and lamivudine therapy

The influence of hepatitis B virus (HBV) genotypes on the natural history and the response to treatment of patients with chronic hepatitis B are of potential interest. Compared to the patients with HBV genotype C, those with genotype B were of a younger age and had a higher cumulative rate of hepatitis B e antigen (HBeAg) seroconversion during the initial 6 years of follow-up. The earlier HBeAg seroconversion in the patients with genotype B, however, did not provide them with a benefit in terms of a reduced risk of developing long-term complications. The response to lamivudine therapy was evaluated in 21 patients infected with HBV genotype B (all of subtype Ba) and 61 with genotype C. There were no differences in the virological response to lamivudine therapy, based on the reduction in median logarithmic HBV DNA titer as well as alanine aminotransferase (ALT) levels, normalization of ALT and the rate of HBeAg seroconversion between the patients with genotypes B and C. No differences were noted either, in the frequency of YMDD mutants at week 52 or the cumulative risk of HBV DNA breakthroughs with YMDD mutations during long-term lamivudine therapy (median 37.5 months). In conclusion, there is no influence of HBV genotypes on the development of long-term complications and lamivudine therapy in Hong Kong.     

B cells and B cell products-helping to restore cellular immunity?

T cells that provide vital protection against tumors, viruses and intracellular bacteria are thought to develop independently of B cells. However, recent discoveries suggest that development of T cells depends on B cells. One way B cells promote T cell development is by providing diverse peptides that may promote positive selection of thymocytes. Diverse peptides and B cells help in diversification of the T cell receptor repertoire and may decrease cross-reactivity in the mature T cell compartment. These new insights may provide the basis for the design of novel therapeutics.     

Comparative Chemotherapeutic Activity of Amphotericin B and Amphotericin B Methyl Ester

The comparative efficacy of amphotericin B and amphotericin B methyl ester (AME) against experimental histoplasmosis, blastomycosis, cryptococcosis, and candidosis in mice was assessed by determining the effect of daily intraperitoneal therapy on 21-day survival and persistence of organisms in internal organs. AME, like amphotericin B, was effective against each of the experimental infections, but the efficacy was lower than the parent compound. For Histoplasma and Blastomyces infections the mean effective dose (ED(50)) of amphotericin B was 0.3 mg/kg, whereas the corresponding values for AME, respectively, were 2.4 and 2.8 mg/kg. For Cryptococcus infection the ED(50) for amphotericin B was 0.2 mg/kg compared with 2.0 mg/kg for AME. The ED(50) of amphotericin B for Candida infection was lower than 0.05 mg/kg and the value of AME was between 0.5 to 0.05 mg/kg. The colony counts from internal organs of the surviving animals after the therapeutic regimens were compatible with the data on survival.     

Acute toxicity of aflatoxin B1 and rubratoxin B in dogs

The effect of ip administrated aflatoxin B1 and rubratoxin B, singly and in combination, on dogs was determined by serum tests, by observations of clinical signs and survival times, and by evaluation of gross and microscopic lesions. The dog is sensitive to the toxic effects of both mycotoxins. Glutamic-oxaloacetic transaminase, lactic dehydrogenase and alkaline phosphatase activities and survival time varied in relation to dose and to the mycotoxin(s) administered. All three plasma enzymes were elevated regardless of dose with the combination of aflatoxin B1/rubratoxin B at 24 hr after dosing, except LDH, which was within the normal range but only at the lowest dose level. Several serum constituents including BUN, cholesterol, uric acid, and total bilirubin were elevated, whereas serum glucose was depressed in dogs treated with the multiple-toxin regimen; these changes were not seen in dogs given only aflatoxin B1 but were characteristic in rubratoxin-treated animals. In general, gross findings at necropsy were similar in all dogs regardless of the dose regimen. A striking similarity existed in the histologic changes observed between lesions experimentally induced by the mycotoxin combination and those lesions reported for dogs fed toxic feed in laboratory studies or in natural cases of hepatitis X. Of particular similarity were the severe kidney lesions observed in dogs exposed to the mycotoxin combination and kidney lesions reported in natural outbreaks of hepatitis X. There can be little doubt of an association between hepatitis X and aflatoxin B1, although it is apparent that the disease probably involves more than a single toxic factor. Our results suggest that hepatitis X in dogs includes aflatoxin B1 as a primary etiological factor but that rubratoxin B also may be involved.     

乙型肝炎病毒B、C基因型全基因组序列的克隆

目的构建乙型肝炎病毒(HBV)B、C基因型全基因组序列克隆。方法从HBV无症状慢性携带者中筛选B、C基因型,提取病毒核酸,设计引物,应用高保真酶对3 200bp的HBV DNA进行全序列扩增,通过克隆技术构建pGEM-HBV重组质粒,测序后进行序列分析。结果获得HBV B基因型和HBV C基因型重组质粒各1株。结论成功构建HBV B基因型和HBV C基因型的全基因组序列克隆,可为进一步研究HBV分子流行病学和基础研究提供工具。     

Amphotericin B tissue distribution in autopsy material after treatment with liposomal amphotericin B and amphotericin B colloidal dispersion

OBJECTIVES: Tissue concentrations of amphotericin B were determined in autopsy material of patients who had been treated with liposomal amphotericin B or amphotericin B colloidal dispersion (colloidal amphotericin B) for suspected or proven invasive fungal infection. PATIENTS AND METHODS: Amphotericin B tissue levels were measured in liver, spleen, lung, kidney, and myocardial and brain tissue of 20 patients who had been treated with lipid-formulated amphotericin B, before they died from multi-organ failure. Seven patients had been treated with liposomal amphotericin B (AmBisome) and thirteen with colloidal amphotericin B (Amphocil). Tissue samples were obtained during routine autopsy, homogenized and extracted with methanol. Amphotericin B concentrations were measured using HPLC after purification by solid phase extraction. RESULTS: The highest amphotericin B levels were found in liver and spleen, followed by kidney, lung, myocardium and brain. In the lung higher amphotericin B concentrations were found after treatment with amphotericin B colloidal dispersion than after therapy with liposomal amphotericin B. CONCLUSIONS: The choice of lipid formulation may influence amphotericin B penetration into the lung.