肝炎患者血清IL—2、IFN—γ和IL—10检测的意义

目的：探讨检测IL－2、IFN－γ和IL－10对病毒性肝炎患者的临床意义。方法：利用ELISA法检测97例病毒性肝炎患者血清IL－12、IFN－γ和IL－10水平，动态观察45例接受免疫增强剂治疗的慢性肝炎患者上述细胞因子的变化，结果：急性肝炎患者血清IL－12及IFN－γ水平平均明显升高（P＜0．01）；慢性肝炎、肝硬化患者血清中IL－10明显高于正常对照组（P＜0．01）。免疫增强剂治疗获得完全应答反应的患者治疗期间血清IL－12、IFN－γ水平明显上升（P＜0．01），IL－10水平下降（P＜0．05）。无应答者治疗过程中上述细胞因子无明显变化。结论：Th1型免疫应答对机体清除病毒起关键作用。Th2型免疫应答与感染慢性化及疾病持续发展有关，免疫治疗可使部分Th2型免疫应答占优势的慢性肝炎患者转化为Th1型占优势。     

ITP血小板特异性抗体和T淋巴细胞亚群及NK细胞变化的意义探讨

目的：检测特发性血小板减少性紫癜（ITP）患者抗血小板膜糖蛋白（GPⅡb／Ⅲa、GPⅠb／Ⅸ）特异性抗体表达、T淋巴细胞亚群及NK细胞的变化,探讨相关因素在ITP发病机制中的作用。方法：应用改良血小板抗原单克隆抗体固相化检测技术（MAIPA）、流式细胞术分别检测52例ITP和24例正常对照组抗血小板膜糖蛋白（GPⅡb／Ⅲa、GPⅠb／Ⅸ）特异性抗体表达、T淋巴细胞亚群及NK细胞变化。结果：ITP组的血小板计数明显低于正常对照组（P〈0．05）;抗GPⅡb／Ⅲa及GPⅠb／Ⅸ抗体A值均高于正常对照组（P〈0．05）;相关分析表明ITP组血小板计数与两种特异性抗体水平均呈负相关关系;在T淋巴细胞亚群变化中,ITP组CD3^＋T淋巴细胞百分比、CD4^＋T淋巴细胞百分比及CD4^＋／CD8^＋的比值均明显低于正常对照组（P〈0．05）,CD8^＋T淋巴细胞百分比则显著高于正常对照组（P〈0．05）;NK细胞百分比明显低于正常对照组（P〈0．05）。结论：血小板特异性抗体及T淋巴细胞亚群的变化可较好地反映ITP这一病理过程,对提高诊断水平及指导临床有一定的实用价值。     

慢性乙型肝炎中医证型与IL—2、IL—10、IL—12、IFN—γ水平的关系初探

目的：探讨慢性乙型肝炎（以下简称慢乙肝）中医证型与血清白细胞介素－2（IL－2）、白细胞介素－10（IL－10）、白细胞介素－12（IL－12）、干扰素－γ（IFN－γ）含量的关系。方法：将50例慢乙肝患者按照中医辨证分型标准分为湿热中阻、肝郁脾虚、肝肾阴虚、瘀血阻络、脾肾阳虚5型。采用双抗体夹心ELISA方法检测血清IL－2、IL－10、IL－12、IFN－γ水平。同时与20例体检健康者的检测水平相比较。结果：除脾肾阳虚型的IL－10和瘀血阻络型的IL－12低于正常对照组外,其余的证型其检测结果均高于正常对照组,P＜0．05;中医辨证分型各组总体差异明显,P＜0．05）;中医证型各组间比较,湿热中阻、肝郁脾虚、瘀血阻铬及脾肾阳虚的IL－2差异明显,P＜0．05,IL－10在各组间无明显差异,肝郁脾虚的IL－12与湿热中阻和肝肾阴虚差异明显,P＜0．05）,肝郁脾虚的IFN－γ与肝肾阴虚、瘀血阻络、脾肾阳虚差异明显,P＜0．05。结论：慢乙肝细胞因子水平与其中医辨认分型有一定的相关性,可作为慢乙肝中医辨证分型的客观依据之一。     

人微小病毒B19感染与小儿ITP的关系

采用PCR检测41例特发性血小板减少性紫瘢（ITP）患儿和22例健康儿童的血清人微小病毒B19（HPVB19）DNA，并用酶联免疫法（ELISA）检测血小板相关抗体。结果ITP组患儿血清中HPVB19DNA阳性率39．0％（16／41）显著高于对照组；ITP组中急性型HPVB19DNA阳性率50．0％（14／28）显著高于慢性型15．4％（2／13）；病毒感染阳性患儿的血小板相关抗体PAIgG、PAIgA明显高于病毒感染阴性患儿。ITP患儿血清中HPVB19DNA阳性率高，病毒感染可导致血小板相关抗体升高而致血小板减少。     

T淋巴细胞PKC在急性ITP发病中的作用

用非同位素标记法检测急性特发性血小板减少性紫癜（ITP）患儿及健康儿童的T淋巴细胞蛋白激酶C（PKC）活性变化，ELISA法检测其血清可溶性白介素2受体（sIL-2R）及PLT计数。结果与健康儿童比较，ITP患儿的T细胞PKC活性及sIL-2R表达显著升高（P〈0．05），PLT计数明显减少（P〈0．01）。PKC活性与sIL-2R表达呈正相关，与PIJT计数呈负相关，sIL-2R表达与PIJT计数呈负相关（P均〈0．05）。提示T淋巴细胞PKC信号转导通路在ITP发生中起重要作用。     

系统性红斑狼疮患者外周血单个核细胞分泌白细胞介素-10 干扰素-γ水平的研究

目的：通过检测系统性红斑狼疮（SLE）患者外周血单个核细胞（PBMC）分泌细胞因子白细胞介素－10（IL－10）、干扰素－γ（IFN－γ）水平，探讨了T淋巴细胞亚群与SLE发病关系。方法：采用酶联免疫吸附法（ELISA）测定30例SLE患者和10名健康志愿者PBMC培养上清IL－10和IFN－γ水平。结论：①SLE活动组PBMC自发分泌IL－10水平明显升高，与非活动组、对照组比较差异有显著性（P＜0．001）；非活动组明显高于对照组（P＜0．01）。②PBMC分泌IFN－γ水平三组（SLE活动组、非活动组和对照组）比较差异无显著性（P＞0．05）。结论：IL－10、IFN－γ作为区分CD4T细胞亚群的指标之一，间接反映了Th1、Th2细胞活化状态。本文研究结果显示，SLE患者PBMC分泌IL－10水平明显增高，且增高程度与病情活动性相关，IFN－γ分泌水平无明显变化。提出Th2型细胞因子介导的免疫应答在SLE的发病机制中占主导地位，即“Th2优势应答”。我们认为PBMC分泌IL－10水平对SLE诊断和病情活动性监测均有一定临床意义。寻找有效方法调节SLE患者体内IL－10水平，从而调节Th1/Th2平衡，将为治疗SLE开辟一条新途径。     

日本血吸虫SjFABPc重组质粒裸DNA免疫小鼠的研究

目的：裸DNA重组质粒pCD-SjFABPc经肌肉注射、皮内途径免疫小鼠,观察其在小鼠体内所诱生的细胞及体液免疫应答。方法：碱裂解法大量制备重组质粒pCD-SjF加强免疫1次。分别于末次免疫后的第20、50、65天共3次用MTT法检测脾脏T淋巴细胞增殖与NK细胞活性,并用双夹心ELISA测定血清细胞因子IL－2、IFN－γ及IL－10含量;ELISA法测定免疫鼠血清IgG抗体。结果：MTT法3次动态测定NK细胞及脾淋巴细胞增殖,不同途径pCD-SjFABPc质粒免疫组均明显高于生理盐水（NS)极空质粒对照组（P＜0．05）。不同途径pCD-SjFABPc免疫组IL－2血清含量的平均值均显著高于NS与空质粒对照组（P＜0．01）;IFN－γ的值也均显著高于NS对照组（P＜0．01）,但与空质粒对照组的差异无显著性（P＞0．05）;不同途径pCD-SjFABPc免疫组IL－10值与NS极空质粒对照组的比较则差异无显著性（P＞0．05）。重组质粒免疫后20d、50d检测,抗体滴度无明显增高;65d检测,pCD-SjFABPc免疫组A值明显高于对照组（P＜0．01）。结论：重组质粒免疫后可诱导小鼠产生有效的细胞与体液免疫应答,且能诱导产生同样类型的细胞因子。     

系统性红斑狼疮患者血小板膜表面糖蛋白的变化

目的 探讨系统性红斑狼疮（SLE）患者血小板膜表面糖蛋白变化，研究血小板膜糖蛋白的变化在该病中的意义。方法 应用流式细胞仪（FCM）测定51例SLE患者血小板膜表面糖蛋白（GP）Ⅱb/Ⅲa和GMP－140的表达水平，并对GPⅡb-Ⅲa明显降低的SLE患者进行血小板聚集功能的测定。结果 SLE患者血小板膜表面GPⅡb-Ⅲa的阳性率明显低于正常对照组（P＜0．01），SLE患者血小板膜表面GMP－140表达水平高于正常对照组（P＜0．01），3例GPⅡb-Ⅲa明显降低的SLE患者存在血小板聚集不良或异常。结论 SLE患者体内产生抗血小板GPⅡb-Ⅲa特异性抗体，导致血小板表面GPⅡb-Ⅲa表达水平降低，并且可能与出血有关。SLE患者体内部分血小板活化，使血小板膜表面GMP－140表达水平增加，这些活化的分子标记物又参与炎症反应，加重原发病。     

特发性血小板减少性紫癜患者治疗前后Th1／Th2水平变化及其临床意义

目的：研究特发性血小板减少性紫癜（ITP）患者药物治疗前后辅助性T淋巴细胞（Th）1/Th2细胞因子水平的变化情况及其临床意义。方法：应用流式细胞仪检测60例ITP患者药物治疗前后干扰素γ（IFN-y）、白细胞介素（IL）-2、IL-4、IL-10水平,分析Th1/Th2细胞因子水平的变化情况,并结合疗效作相关性分析。以60名健康体检者为对照。结果：药物治疗有效的ITP患者治疗后Th1细胞因子IL-2、IFN-1水平较治疗前及对照组均显著升高（P〈0．01）。而Th2细胞因子IL-4及IL-10水平在3组间均无显著性差异（P＞0．051．Th1/Th2比例升高。ITP患者Th1细胞因子IL-2、IFN．叮水平在治疗显效组表达均明显高于良效及进步组和无效组（P〈0．05）,良效及进步组与治疗无效组比较亦有显著性差异（P〈0．05）;经相关性分析,患者的疗效与IFN-γ、IL-2均呈正相关。结论：ITP患者药物治疗后Th1细胞因子IL-2、IFN-γ表达升高,且Th1与ITP患者疗效密切相关。     

系统性红斑狼疮患者血浆11—去氢—血栓烷B2测定及其临床意义

目的 观察系统性红斑狼疮（SLE）患者体内血小板活化状态。方法 采用酶联免疫吸附法（ELISA）检测32例SLE患者血浆11－去氢－血栓烷B2（DH－TXB2）,同时测定TXB2,并与对照组比较。结果 与对照组比较,SLE患者血浆DH－TXB2、TXB2均显著增高（P＜0．01及P＜0．05）,且DH－TXB2增高程度明显高于TXB2,对对照组无重叠,肾受累者显著高于无肾脏受累者（P＜0．05）,经治疗者显著低于初发未治疗者（P＜0．01）,但仍高于对照组（P＜0．05）。血浆TXB2与DH－TXB2浓度与疾病活动指数均呈正相关（r1＝0．478,P＜0．05及r2＝0．530,P＜0．01）。结论 血浆DH－TXB2水平测定是准确反映血小板活化的指标,SLE患者体内血小板高度活化。     

TH1 and TH2 cell polarization increases with aging and is modulated by zinc supplementation

Elderly subjects suffer from increased levels of activated T cells and a TH1/TH2 imbalance. Zinc deficiency of the aged is correlated with decreased cell-mediated immune responses. The association of age and zinc adjustment with the amounts of TH1 (CCR5+) and TH2 (CCR4+) cell populations in healthy aged old donors enrolled in the European ZINCAGE project was examined. Old and nonagenarian individuals revealed increased TH1, TH2 cell numbers and a decreased TH2/TH1 ratio in comparison to young individuals. The differences between TH2/TH1 ratios of young and old/nonagenarians arose from young females. Adjusted zinc status led to enhanced TH2 and TH1 amounts in fresh whole blood and thawed cells of aged donors whereas increased HLA-DR+ expression and a generally lower CCR5 expression was observed on thawed PBMC. In conclusion, aging is associated with an increase in T helper cell polarization, and changes in TH2/TH1 subsets are more obvious in women than in men. Advanced healthy aging is accompanied by TH cell polarization, too. Moderate zinc supplementation in vivo alters TH proportions. Longer zinc treatment will give more insight into the beneficial effect of zinc on T helper cell modulation.     

幼年小鼠应用抗生素致免疫反应亢进的研究

目的　探讨卡那霉素如何影响 Th1 / Th2 平衡 ,使 Th2 免疫反应占优势。方法　 BAL B/ c雄性小鼠 1周龄、5 0周龄 ,且每组 15只 ,其前未用过抗生素 ,然后分别让 1周龄组 ,5 0周龄组 ,口服硫酸卡那霉素 6 0 0 mg/ d,连用7天 ,在服用硫酸卡那霉素停止后的第 10周将小鼠杀掉 ,取眼球血测 Ig E、Ig G1 、Ig G2 a,并做脾的单个核细胞分离 ,在体外用伴刀豆蛋白 A2 .5 ug/ ml(Sigma公司 )刺激约 96小时后测上清液中的 IL - 4及 IFN- r。结果　 5 0周龄比 1周龄组脾脏单个核细胞分泌的 IL - 4量明显升高 ,IFN- r量明显降低 (P均 <0 .0 5 ) ,并伴随着血清中 Ig E、Ig G1 明显升高 ,Ig G2 a明显降低 (p<0 .0 5 )。结论　该结果显示在幼年时应用抗生素能影响 Th1 / Th2 平衡 ,使 Th2 反应亢进     

Effect of improved zinc status on T helper cell activation and TH1/TH2 ratio in healthy elderly individuals

Mild zinc deficiency is a common condition in healthy elderly individuals leading to impaired cell-mediated immune response. Here we report the effect of improved zinc status on TH1/TH2 balance and on the activation status of T helper cells in 19 healthy elderly subjects aged 69.8 ± 5.1 years. Our investigations revealed a mild zinc deficiency which was adjusted by oral zinc supplementation for seven weeks. Improved serum zinc levels significantly reduced levels of activated T helper cells whereas changes in TH1/TH2 ratio (determined by CCR4 and CCR5 expression) were not observed. These findings suggest that elderly individuals may benefit from moderate zinc supplementation due to improved immune response leading to reduced incidences of autoimmune diseases and infections.Presented at the Zinc Age Conference, Madrid, Feburary 10–13, 2006.     

TH17细胞平衡及免疫微环境在过敏性哮喘发病中的作用

目的探讨辅助性T细胞17(TH17)及其相关细胞因子表达水平在过敏性哮喘发病中的作用机制。 方法选取120例哮喘患者作为本文的研究对象。根据患者的哮喘类型,将120例患者分为过敏性哮喘以及非过敏性哮喘两组,每组60例患者,同时选择同期入院体检的48例健康人群作为对照组。测定三组受试者外周血中Th 17、Treg、Th2比例以及血浆和诱导痰中IL-17A、IL-4、IL-5、IL-10及IL-18水平。此外测定三组患者诱导痰中中性粒细胞、嗜酸粒细胞比例。 结果过敏性哮喘组Th17比例高于对照组(P<0.05);过敏性哮喘组Treg比例低于对照组(P<0.05);过敏性哮喘组Th17比例低于非过敏性哮喘组(P<0.05);三组间Th2比例无统计学意义(P>0.05)。过敏性哮喘组IL-17A、IL-4以及IL-5水平高于对照组(P<0.05);过敏性哮喘组IL-10及IL-18水平低于对照组(P<0.05),过敏性哮喘组IL-17A水平低于非过敏性哮喘组(P<0.05);过敏性哮喘组IL-4水平高于非过敏性哮喘组(P<0.05),过敏性哮喘组IL-17A、IL-4、IL-5、IL-10及IL-18水平均高于对照组。过敏性哮喘组IL-17A以及IL-18水平低于非过敏性哮喘组(P<0.05);过敏性哮喘组IL-4水平高于非过敏性哮喘组(P<0.05);两组间IL-5、IL-10水平无统计学意义(P>0.05)。过敏性哮喘组中性粒细胞、嗜酸粒细胞比例均高于对照组(P<0.05),过敏性哮喘组中性粒细胞比例低于非过敏性哮喘组,且差异存在统计学意义(P<0.05);过敏性哮喘组嗜酸粒细胞比例高于非过敏性哮喘组(P<0.05)。 结论TH17及其相关细胞因子的免疫失衡与过敏性哮喘的发生密切相关。     

Helper (TH1, TH2, TH17) and regulatory cells (Treg, TH3, NKT) in rheumatoid arthritis

The immune response foreign antigens require a perfect coordination of cells that participate in its different phases. The objective of the response is the rapid destruction of the microorganisms with a minimum repercussion on self-cells and tissues. The regulation of this process is carried out fundamentally by T lymphocytes. There are two main types of coordinator cells: helper cells, what organize the initial immune response, and regulatory cells, what avoid immune attack against self and once the infection is controlled, disassemble the response. There are three types of helper cells which coordinate answers to intracellular parasites (TH1), helmints (TH2) and extracellular bacteria and fungi (TH17). The hyperfunction of TH17 cells is associated with diseases as reumatoid arthritis, due to the hypersecretion of the proinflammatory citoquine IL17. The condition of helper or regulatory cell is the current object of review. TH1, TH2 and TH17 cells have helper and also regulatory functions. In addition, regulatory T cells play an important role in the coordination of the first moments of the response to viral infection in a direct and indirect way, inducing differentiation of TH17 cells.     

Detection of T lymphocyte subsets of children with Helicobacter pylori infection

AIM: To study the transformation of T lymphocyte subsets in children with Heliobacter pylori (H pylori) infection. METHODS: The H pylori infection status were determined by a combination of ELISA and Western blot (immunoblot) technique in 98 children and T lymphocyte subsets in peripheral blood were determined by flow cytometrical analysis. RESULTS: There were 75 children positive with H pylori infection and 23 negative in 98 children. Comparing the proportion of peripheral blood T lymphocytic subsets in children with H pylori infection and without H pylori infection, it was found that a higher proportion of CD4 T-cells in infected children (39.02±7.71 VS 34.25±10.73, t=2.246,P<0.05) and higher value of CD4 to CD8 T-cells ratio (1.51±0.52 VS 1.25, t=2.104, P<0.05) were present, but there were not significant differences in CD3 T-cells and CD8 T-cells (73.11±10.02 VS 69.49±17.08, 27.22±6.07 VS 28.27±8.67, P>0.05). CONCLUSION: Th1 cell-mediated immune responses may be induced by H pylori infection in children.     

TH1 and TH2 Cytokine Production Among Asthmatic Children After Immunotherapy

Allergen immunotherapy results in a number of changes in clinical and immunological parameters. To study the kinetic influence of immunotherapy on cytokine production, we evaluated the ratios of Th1/Th2 for patients receiving mite-extract immunotherapy with 3-month intervals. Changes in Th1/Th2 ratio were calculated based on the intracellular cytokine and surface CD4 staining of peripheral blood mononuclear cells (PBMC). No significantly different Th1/Th2 ratios were detected after immunotherapy, although ratios were lower for asthmatic children when compared with normal controls. Cytokine production was also determined on supernatants from mite- or mitogen-activated PBMC cells. A significantly increased production of all cytokines was detected from activated PBMC from patients after immunotherapy. Thus hyposensitization with mite extract for 1 year might improve the cytokine production capacity of asthma patients' PBMC.     

TH1 and TH2 Cytokine Production Among Asthmatic Children After Immunotherapy

Allergen immunotherapy results in a number of changes in clinical and immunological parameters. To study the kinetic influence of immunotherapy on cytokine production, we evaluated the ratios of Th1/Th2 for patients receiving mite-extract immunotherapy with 3-month intervals. Changes in Th1/Th2 ratio were calculated based on the intracellular cytokine and surface CD4 staining of peripheral blood mononuclear cells (PBMC). No significantly different Th1/Th2 ratios were detected after immunotherapy, although ratios were lower for asthmatic children when compared with normal controls. Cytokine production was also determined on supernatants from mite- or mitogen-activated PBMC cells. A significantly increased production of all cytokines was detected from activated PBMC from patients after immunotherapy. Thus hyposensitization with mite extract for 1 year might improve the cytokine production capacity of asthma patients' PBMC.