CpG寡脱氧核苷酸增强免疫抑制小鼠对乙肝疫苗的免疫应答

目的 :探讨CpG寡脱氧核苷酸 (ODN)增强免疫抑制小鼠对乙肝疫苗的免疫应答效果。方法 :选用环磷酰胺 (CTX)所致免疫抑制模型小鼠 ,以乙肝疫苗和 2 0 μgCpGODN联合或单独左胫前肌肌注免疫C5 7BL/6小鼠。 2wk后以同样剂量加强免疫 1次 ,再过 3wk后摘除眼球取血 ,用ELISA法检测抗 HBsIgG抗体和IL 12的水平。同时 ,无菌取脾脏做HE染色 ,观察脾脏淋巴细胞的数量和细胞核的变化。结果 :CpGODN与疫苗联合注射组产生的绝对抗体量比单独注射疫苗组提高 1倍 ;注射组产生IL 12的水平较单独注射疫苗组也有明显升高。光镜下各组脾脏淋巴细胞的变化如下 :正常对照组脾脏可见大量的淋巴细胞 ;与正常对照组相比较 ,CTX组的淋巴细胞明显稀少 ;而CTX加CpG组的淋巴细胞数明显增多 ,细胞核也明显增大。结论 :CpGODN能增强免疫抑制小鼠对乙肝疫苗的免疫应答     

铝佐剂吸附CpG ODN增强重组乙肝疫苗免疫效应

目的探索新佐剂CpG ODN在重组乙肝疫苗中的免疫效应及吸附状态。方法 CpG ODN佐剂与重组乙肝疫苗混合免疫小鼠,收集小鼠血清检测。通过免疫小鼠后的体液和细胞免疫应答水平来评价双佐剂乙肝疫苗对重组乙肝疫苗的增强作用。CpG ODN佐剂与乙肝疫苗在不同温度条件下混合吸附,计算在不同吸附时间下CpG ODN佐剂在乙肝疫苗中的吸附率。结果CpG ODN佐剂可以提高重组乙肝疫苗在体液及细胞中的免疫效果。CpG ODN佐剂在室温条件下1 h即可在乙肝疫苗中达到吸附平衡,于2~8℃放置5 d的条件下其吸附率约在50%左右。结论 CpG ODN能够增加乙肝疫苗的免疫应答,其免疫效果可提高5倍以上。     

重组IL-12对乙肝疫苗在小鼠诱导免疫应答的作用

目的:观察重组IL-12对乙肝疫苗在小鼠诱导应答的强度与性质的作用,探讨将重组IL-12用作乙肝治疗性疫苗分子佐剂的可能性.方法:将乙肝疫苗联合重组IL-12肌注免疫小鼠,检测小鼠产生的抗乙肝表面抗原特异性体液和细胞免疫应答.结果:乙肝疫苗联合重组IL-12能明显增强小鼠T淋巴细胞的增殖活性、促进分泌细胞因子IFN-γ和IL-2并提高IgG2a抗体水平.结论:重组IL-12可显著增强乙肝疫苗诱导的细胞免疫应答,并使免疫应答转向Th1型.     

CpG ODN的体外免疫活性及其作为乙肝疫苗佐剂的研究新进展

为了增强乙肝疫苗的免疫效力,开发研制新型疫苗佐剂是当前国内外学者研究的热点之一。近年来的研究表明：含CpG基序的寡脱氧核苷酸（CpG ODN）在体外实验中,对人和鼠的多种免疫细胞有活化作用;在动物实验中,CpG ODN不仅可增强乙肝疫苗的体液免疫应答,还可增强其细胞免疫应答,这正是铝佐剂所达不到的,且CpG ODN与铝佐剂具有协同效应,今后可考虑双佐剂联合乙肝疫苗接种,能提高接种人群的应答率,刺激机体产生更有效的保护作用;动物实验还显示,CpG ODN能增强HBV转基因小鼠对乙肝疫苗的免疫应答和抗病毒效应,说明乙肝疫苗辅以CpG ODN可望作为临床上免疫治疗慢性HBV感染的可行性途径;Ⅰ期临床试验表明CpG ODN对人体较安全。     

CpG ODN联合乙型肝炎疫苗与乙型肝炎疫苗免疫效果对比研究

目的探讨CpG ODN乙型肝炎疫苗与市售乙型肝炎疫苗对Balb/c小鼠的免疫作用效果。方法将乙肝疫苗、乙肝疫苗+100μgCpG ODN分别肌肉注射到4～6周龄,16～18g Balb/c小鼠体内,于第1次免疫后28d以同样剂量加强免疫1次。分别于第1次免疫后28、42、63 d收集小鼠血清,用ELISA方法检测抗HBs IgG抗体。结果实验组产生的抗HBs IgG与对照组HBsIgG相比P<0.05,具有显著性差异。28、42、63 d抗HBs IgG分别是对照疫苗的114倍、4.73倍、5.41倍以上。结论 CpG ODN能够显著增强小鼠对乙肝疫苗的免疫应答,在较短时间内显著的提高乙肝抗体的产生水平,免疫效果是乙肝疫苗的5倍以上。     

CpG ODN对rHBsAg免疫小鼠淋巴细胞增殖反应的影响

目的：进一步探讨以合成含CpG基序的寡核苷酸（CpG ODN）作为免疫增强剂与重组乙型肝炎表面抗原（rHBsAg）联合免疫小鼠,观察其淋巴细胞增殖反应效应.方法：经后腿胫骨前肌初次免疫BALB/c小鼠两周后加强免疫1次;MTT和3H-TdR掺入法分别检测免疫小鼠胸腺及脾脏淋巴细胞增殖反应.结果：加CpG ODN组与单纯注射抗原及疫苗组相比,淋巴细胞增殖反应显著增强.而且,除胸腺特异性增殖中的CpG加疫苗组和疫苗组外,其他各实验组与对照组相比,淋巴细胞增殖反应也都显著增强.结论：CpG ODN能够明显刺激小鼠胸腺及脾脏淋巴细胞增殖反应,显示出强而有效的免疫增强效应,有望成为一种新型的免疫佐剂.     

CpG ODN对ZP~(121-140)表位合成肽诱导的免疫应答和免疫避孕效应影响

目的:探讨CpG ODN对透明带(四)抗原表位ZP~(121-140)合成肽诱导的免疫应答和免疫避孕效应影响.方法:应用人工合成ZP2~(121-140)表位肽,与20μg CpG ODN或等量完全弗氏佐剂(CFA)混悬后左胫前肌免疫雌性BALB/c小鼠,初次免疫后2、4、6周各加强免疫一次,共免疫四次.在每次加强免疫前和末次免疫后两周断尾取血,分离血清,分析特异性lgG抗体及非特异性细胞因子IFN-γ、TNF-Ⅱ,IL-10水平;收集小鼠阴道冲洗液,离心取上清,分析特异性IgA抗体水平.免疫小鼠生育实验结束后,取卵巢组织行病理学分析.结果:CpG ODN组诱导的特异性IgG和IgA水平较CFA组有增高趋势,但无显著性差异(P＞O.05).CpG ODN组诱导的非特异性IFN-γ和TNF-水平明显高于CFA组(P＜0.05);而CpG ODN组诱导的IL-10水平显著低于CFA 组(P＜0.05).两种佐剂对小鼠的受孕率影响没有显著性差异,但CpG ODN组孕鼠每胎产仔数明显低于CFA组(P＜O.05).病理学分析显示实验小鼠卵巢组织无病理性变化.结论:CpG ODN对Zp~(121-140)合成肽诱导的免疫应答和免疫避孕效应略优于CFA,更适合用于避孕疫苗佐剂研究.     

CpG ODN佐剂对乙肝疫苗抗原免疫原性的影响

目的探究CpG ODN佐剂对不同抗原含量乙肝疫苗免疫效果的影响。方法在体液免疫方面不同抗原浓度的CpG ODN与Al(OH)3佐剂乙肝疫苗免疫Balb/c小鼠,于免疫后第2、4、6、8、10周收集血清,检测小鼠体内相对效力ED50和血清中的anti-HBs抗体水平;在细胞免疫方面测定诱导产生IFN-γ水平及IgG2a应答水平。结果 CpG ODN与Al(OH)3佐剂可以有效协同HBs Ag诱导机体产生的抗体滴度达49 427 mIU/ml,抗体效价随时间延长而增加。乙肝抗原减半后双佐剂乙肝疫苗诱导机体产生的特异性抗体滴度可达41 225 mIU/ml,高于无CpG ODN佐剂的铝佐剂疫苗。在诱导细胞分泌方面,对照组诱导产生IFN-γ水平及IgG2a应答水平明显低于所有双佐剂疫苗组。结论 CpG ODN对HBsAg具有良好佐剂活性,并与铝佐剂有协同作用,二者联合应用可以降低乙肝抗原用量并提高疫苗免疫原性。     

多房棘球绦虫ELP重组蛋白疫苗免疫Balb/c小鼠引起的免疫应答

目的：观察多房棘球绦虫ELP重组蛋白疫苗免疫Balb／c小鼠引起的体液和细胞免疫应答。方法：在成功构建多房棘球绦虫elp基因原核表达载体(pQ-ELP)的基础上，以亲和层析法纯化重组蛋白，SDS-PAGE鉴定，Bradford法进行定量。用ELP重组蛋白(A组)及ELP重组蛋白加弗氏佐剂(B组)免疫Balb／c小鼠(10只／组)，以生理盐水(NS组)作对照。ELISA方法检测免疫后不同时间产生的特异性IgG1，IgG2a和IgG2b水平。双抗体夹心ELISA试剂盒检测免疫鼠脾脏单个核细胞(PMNC)在受到特异抗原刺激后，产生IL-4、IL-12和IFN-γ的能力，^3H-TdR掺入法检测脾淋巴细胞的增殖能力。结果：本研究成功获得高纯度重组蛋白ELP。首次免疫后2周，A、B组小鼠均产生了大量的特异IgGl抗体，B组还产生了少量的特异IgG2b抗体。除B组小鼠脾PMNC产生了微量IFN-γ外，A组、NS组IFN-γ及各组IL-4和IL-12均未检出。A、B组小鼠脾淋巴细胞增殖能力增高，其淋巴细胞CPM值分别为NS组的2．8和10．8倍，受到多房棘球绦虫抗原或刀豆素刺激时，A、B组淋巴细胞增殖更明显。结论：多房棘球绦虫ELP重组蛋白疫苗可引起很强的体液免疫应答和微弱的细胞免疫应答，如与弗氏佐剂联合应用引起的细胞免疫应答会进一步提高。     

基因佐剂在DNA疫苗中的应用

基因佐剂可将编码某些具有佐剂作用成分的基因与DNA疫苗共免疫，在体内表达相应的蛋白，增强T、B细胞对疫苗成份的体液及细胞免疫应答。目前研究较多的基因佐剂有：细胞因子基因佐剂如IL-12、IL-2、IFN—γ、GM-CSF等；共刺激分子基因佐剂如B7—2、CD40L等；免疫刺激DNA序列基因佐剂如CpG、ODN等。基因佐剂在体内与疫苗基因共表达，改善抗原的周边微环境，提高疫苗的免疫效果。因此，基因佐剂是目前DNA疫苗研究中的热点问题之一。     

Development in in vitro toxicology

There are three principal pressures driving the development of in vitro toxicology: (1) the need for more efficient testing systems to cope with the large number of xenobiotics currently being developed; (2) public pressure to reduce animal experimentation; and (3) a need for a better understanding of the mechanisms of toxicity. Within this, in vitro toxicology is focused on local, systemic, and target-organ toxicity. It is becoming increasingly apparent that a step or decision-tree approach using input of a variety of experimental data (physicochemical properties, biokinetics, cytotoxicity) provides the most efficient system for predicting toxicity. Examples of the use of in vitro toxicity systems for prediction of systemic toxicity and target-organ (liver) toxicity are presented.Originally presented at ECCP 93.     

Seasonal variations in acute toxoplasmosis in pregnant women in Slovenia

Between December 1999 and December 2004, 40 081 pregnant women were examined for toxoplasmosis with Toxo-IgG, Toxo-IgM enzyme immunoassay. Women with positive results were then retested with the Toxo-IgG avidity assay for recent toxoplasmosis. Recent acute toxoplasmosis in pregnant women was found to be significantly more frequent (p < 0.01) during winter than summer. The incidence of acute toxoplasmosis during winter-spring was also significantly more frequent (p < 0.025) than summer-autumn. This phenomenon should be taken into account when formulating preventive measures for toxoplasmosis, especially for pregnant women.     

Age-related changes in polyamines in memory-associated brain structures in rats

Polyamines putrescine, spermidine and spermine are positively charged aliphatic amines and have important roles in maintaining normal cellular function, regulating neurotransmitter receptors and modulating learning and memory. Recent evidence suggests a role of putrescine in hippocampal neurogenesis, that is significantly impaired during aging. The present study measured the polyamine levels in memory-related brain structures in 24- (aged), 12- (middle-aged) and 4- (young) month-old rats using liquid chromatography/mass spectrometry and high performance liquid chromatography. In the hippocampus, the putrescine levels were significantly decreased in the CA1 and dentate gyrus, and increased in the CA2/3 with age. Significant age-related increases in the spermidine levels were found in the CA1 and CA2/3. There was no difference between groups in spermine in any sub-regions examined. In the parahippocampal region, increased putrescine level with age was observed in the entorhinal cortex, and age did not alter the spermidine levels. The spermine level was significantly decreased in the perirhinal cortex and increased in the postrhinal cortex with age. In the prefrontal cortex, there was age-related decrease in putrescine, and the spermidine and spermine levels were significantly increased with age. This study, for the first time, demonstrates age-related region-specific changes in polyamines in memory-associated structures, suggesting that polyamine system dysfunction may potentially contribute to aged-related impairments in hippocampal neurogenesis and learning and memory.     

休克过程中肾上腺髓质素变化的研究进展

Adrenomedullin (AM) is a new peptidergic regulator of vascular function. AM serves as a hormone, which has many biological properties, plays an important role in the many pathophysiological processes, especially shock. This review will highlight the structure, biological properties of AM and the relationship between AM and shock.     

Secular change in menarche in women in Madrid

The age at menarche was estimated by recollection in 1617 women between the ages of 18 and 60 in Madrid and a nearby suburb, Pinto. The population of Pinto is working-class and the Madrid group, taken from residential neighbourhoods , belongs to the upper middle class. In both groups we found a diminution in average age at menarche, from 14.04 to 13.02 years in Madrid and from 14.55 to 13.16 years from about 1935 to about 1965 in Pinto. These changes have been more intense in the group which is less well-off economically, where living conditions have varied much more drastically.     

Uteroglobin in the developing rabbit conceptus in vivo and in vitro

Summary Uteroglobin (UGL) was measured in day- 4 to day-10 rabbit conceptuses by a competitive ELISA. Levels in blastocyst fluid, tissues, coverings and in the early fetus were determined separately. The total amount of UGL increased from 18.4 ng to 6.8 g per conceptus. The UGL content of individual day-6 blastocysts was studied in vitro. Culturing was carried out up to 60 h in Ham's F10 medium with polyvinylpyrrolidone as macromolecular component, with and without progesterone, and with progesterone plus estradiol. UGL was determined in the blastocyst fluids, tissues with coverings and in the culture media. After labelling with [³⁵S]-methionine, protein patterns of total blastocysts and of culture media were analysed by two-dimensional gel electrophoresis and fluorography. The morphology of cultured blastocysts was examined by electron microscopy. During 60 h of culture, the blastocysts expanded in diameter by 84%, and released 19% of their initial UGL content into the medium, independent of the hormonal substitution. Neither de novo synthesis, nor degradation of UGL was found: the protein remained unlabelled in fluorography, and its total quantity was not significantly different from that of non-cultured controls. Trophoblast, endoderm and embryoblast cells showed well preserved cell organelles and intercellular junctions, while the morphological differentiation of the germ layer was inhibited.