鹦鹉热衣原体重组主要外膜蛋白诱导小鼠免疫应答的观察

用鹦鹉热衣原体 (Chlamydiapsittaci,Cps)重组主要外膜蛋白 (RecombinantMajorOuter Membraneprotein ,r MOMP)免疫小鼠 ,观察小鼠免疫后对r MOMP和Cps菌体蛋白的免疫应答。r MOMP皮下注射免疫BALB/c小鼠 ,对照组仅注射佐剂。免疫前和第 3次免疫后 10d收集血清 ,以常规ELISA法检测抗体效价 ;MTT方法检测脾淋巴细胞对r MOMP和Cps菌体蛋白的特异性增殖反应。免疫组小鼠在免疫 38d后免疫血清中抗r MOMP的抗体效价可达 1∶2× 10 4,抗Cps菌体蛋白的抗体效价为 1∶4× 10 3 ;脾淋巴细胞对r MOMP和Cps菌体蛋白的增殖指数明显高于佐剂对照组 (p <0 .0 1,具有显著性意义。r MOMP在小鼠体内可诱导Cps特异性的体液免疫和细胞免疫应答 ,说明具有较好的免疫原性     

乙肝病毒RNA复制子疫苗与DNA疫苗对小鼠免疫效率的比较

为建立并获取更有效的乙肝疫苗,本实验通过将所构建的HBV RNA复制子疫苗和DNA疫苗分别免疫小鼠,检测细胞免疫与体液免疫的效果.结果表明,以pSFV为基础构建的疫苗载体免疫小鼠后采集的血清中抗体效价不随免疫剂量的增加而提高,在较低剂量免疫的时候,RNA复制子疫苗所产生的抗体效价优于DNA疫苗.并且RNA复制子疫苗在以较低剂量免疫后脾细胞CTL活性高于DNA疫苗.本研究证明HBV RNA疫苗比DNA疫苗表达效果更好,安全性更高,更具有应用前景.     

抗HBV与HCV免疫乳的实验免疫研究

确定重组HBV表面抗原和HCV核心抗原对奶牛的最适免疫剂量、免疫次数及间隔时间,观察牛奶中抗HBV与抗HCV的效价与消长规律。表达HBV表面抗原和HCV核心抗原的重组菌经IPTG诱导后,目的蛋白以包涵体形式存在。放大发酵工艺,菌体发酵密度达40g／L,目的蛋白表达量为26％～30％。将包涵体变性、复性后,测定蛋白含量,并用SDS-PAGE鉴定。采用不同的剂量和不同的抗原处理方法免疫奶山羊和奶牛,检测HBV抗体、HCV抗体、干扰素与白介素等细胞因子。免疫奶山羊羊奶中的HBV抗体效价最高为1：80,HCV抗体效价最高为1：20;6个月后,HBV抗体效价无明显下降,HCV抗体效价下降明显。对奶牛5个批次的免疫结果显示,免疫次数应多于3次,免疫剂量以每头牛300μg为宜,间隔时间以1个月为宜。免疫牛奶中HBV抗体的阳性率约为66．0％,抗体效价最高为1：160;HCV抗体阳性率约为17．0％,维持时间较短;孕牛比旺奶牛产生抗体的效价高。免疫牛奶中检测到了干扰素和白介素等细胞免疫活性因子。     

银鲫口服嗜水气单胞菌疫苗的免疫和免疫组化研究

室内恒温条件下(水温24±1℃)分别用微胶囊疫苗与全细胞疫苗口灌免疫银鲫,口灌免疫后每周取血和前、中、后肠肠黏膜,间接ELISA检测其抗体效价。结果发现:微胶囊疫苗组鱼血清抗体效价较高,峰值为1∶80,且维持时间较长;各肠段的黏膜抗体中,后肠黏膜抗体效价最高,微胶囊疫苗组峰值达1∶320,前肠的肠黏膜抗体效价也有一定的水平为1∶40;肠道免疫组化试验也得到相似结果:在不同时间对前、中、后肠切片进行免疫组化染色,从光镜下观察到肠黏膜外层和黏膜下层都有阳性着色,以包裹全菌苗为最多,未包裹菌苗较少。同时进行两组疫苗的池塘网箱免疫试验,结果表明:微胶囊疫苗组与全细胞疫苗组的相对百分成活率分别为61.1%和50%,微胶囊疫苗组略高于全细胞疫苗组;口服免疫后两组鱼血清中均能产生较高的抗体效价,在30d左右抗体效价最高达1∶512,能维持较长时间约8—10个月,但两者差异不显著。     

绿幕安的临床应用

绿幕安（宝泰ＰＭ免疫调节剂）是具有广泛交叉免疫和广谱抗感染作用的绿脓杆菌（ＭＳＨＡ）菌毛株菌苗。绿幕安是经现代生物工程培育,由低毒的绿脓杆菌菌体,嫁接上具有广谱免疫的菌毛,利用其广谱、低毒、较强免疫原性进行全身非特异性减敏治疗,刺激机体产生广谱抗体,...     

汉坦病毒嵌合基因 G2S0.7在昆虫细胞中融合表达产物的免疫学研究

在前期工作基础上,对汉坦病毒糖蛋白(GP)和核蛋白(NP)的嵌合基因G2S0．7表达产物的免疫学特性进行进一步的研究。将汉坦病毒含有G2S0．7嵌合基因的重组杆状病毒在昆虫细胞中融合表达并免疫BALB／c小鼠,用ELISA、微量细胞培养中和实验及淋巴细胞增殖实验检测免疫应答效果,以研究嵌合基因的免疫效果。结果表明,用该融合蛋白免疫小鼠,可诱导产生抗汉坦病毒NP及GP特异性的抗体,抗体效价分别为1：3200及1：200;同时融合蛋白还可刺激机体产生低水平的中和抗体和明确的淋巴细胞增殖反应。说明汉坦病毒G2S0．7嵌合基因表达的融合蛋白既可刺激机体产生特异性的抗汉坦病毒体液免疫应答,也可刺激机体产生特异性的细胞免疫应答,为进一步进行汉坦病毒基因工程疫苗的研究奠定了实验基础。     

抗体跨越血脑屏障去攻击病毒

约翰·霍普金斯大学医学院(Baltimofe,MD)声称采用单克隆抗体阻止了病毒在大鼠神经元培养中的繁殖。该校的Diane Griffin及其同事说,这些结果证实了如下结论:是体液免疫,而不是细胞免疫,阻止了病毒在SCID患鼠脑中的感染。这就提示可以用单抗治疗脑部的病毒感染。机体抗病毒感染有两条途径:体液免疫(抗体与病毒蛋白的结合)及细胞免疫(T细胞攻击受感染的组织细胞)。体液免疫对于组织外的体液很重要,但人们认为,要从组织(包括脑)中清除病毒,则需要细胞免疫。     

基因枪法和肌内注射法免疫HCV NS3/4A诱导小鼠免疫应答的比较

目的比较肌内注射法和基因枪法对免疫效果的影响。方法将携带HCV NS3/4A基因的质粒pcDNA3.1-NS3/4A,分别肌内注射和基因枪免疫BALB/c小鼠。基因枪免疫采用2μg/只和10μg/只两种剂量,肌内注射免疫的剂量为100μg/只,共免疫3次。分别于第3次免疫后10 d和20 d,收集小鼠血清、分离脾细胞;ELISA法检测小鼠血清中特异性抗体水平;ELIspot检测小鼠IFN-γI、L-2和IL-4;非放射性细胞毒性试验检测其对肿瘤细胞的杀伤作用,从而比较肌内注射和基因枪法激发小鼠产生体液免疫和细胞免疫应答的影响。结果免疫后第10 d检测结果表明,2μg/只和10μg/只基因枪法免疫产生的抗体效价分别为1∶500和1∶1 000,100μg/只肌内注射免疫产生的抗体效价为1∶1 000。第20 d再次检测,基因枪法产生的抗体效价仍为1∶500和1∶1 000,肌内注射降低为1∶500。2μg/只和10μg/只基因枪免疫产生的IFN-γ和IL-2均高于100μg/只肌内注射免疫。但这两种免疫方法均未激发IL-4的水平。在不同的效靶比情况下,10μg/只和2μg/只基因枪免疫激发的CTL细胞杀伤活性均高于100μg/只肌内注射免疫。结论基因枪法免疫小鼠诱导的体液免疫和细胞免疫应答均显著高于肌内注射免疫。     

FMDV VP1基因与HSP70基因在毕赤酵母中的融合表达及免疫特性

利用毕赤酵母系统对O型口蹄疫病毒VP1基因与结核杆菌HSP70基因进行融合表达,并检测此融合蛋白对小鼠细胞免疫和体液免疫的影响。将人工合成的O型口蹄疫病毒VP1基因与结核杆菌HSP70基因克隆入酵母表达载体pPICZαA中,以电穿孔法转化酵母菌X-33,用Zeocin YPDS平板筛选重组子,经甲醇诱导表达后,SDS-PAGE和免疫印迹分析表达产物。以皮下接种的方式给小鼠进行3次免疫,同时设两组对照,分别免疫PBS和常规灭活疫苗,然后通过MTT法和ELISA分别检测淋巴细胞的增殖情况和抗体水平。结果表明融合蛋白既能诱导细胞免疫应答又能诱导体液免疫应答,其诱导产生的抗体水平略低于常规灭活疫苗,而细胞免疫水平则高于后者。     

O139霍乱灭活全菌体菌苗免疫后人群抗体水平测定

Ｏ１３９霍乱弧菌经甲醛灭活制备而成的全菌体菌苗在广西田阳县进行临床考核。每组５０名中学生经肌肉途径接种４５亿菌和９０亿菌两种剂量,一个月后８０％接种者血清中杀弧菌抗体升高,最高可达１∶１６０,免疫后３个月开始缓慢下降,但仍有６８－７２％接种者抗体效价在１∶２０以上。高剂量组半数接种者的杀弧菌抗体可维持存在６个月。血清中凝集抗体升高明显,但３个月后下降也明显。与对照组Ｏ１菌苗相比,两种抗体水平升高与下降趋势一致。采用ＥＬＩＳＡ法测定抗毒抗体,Ｏ１３９菌苗组与Ｏ１菌苗组结果相似,均有一定上升。上述结果提示Ｏ１３９霍乱灭活菌苗可作为短效应急菌苗使用     

Immunosuppressive effect of cyclosporin A on insect humoral immune response

Cyclosporin A suppressed humoral immune response of Galleria mellonella larvae. Insects were immunized with LPS Pseudomonas aeruginosa and then injected with cyclosporin A. Immunosuppressive effects were expressed both, in larvae treated with cyclosporin A at the initial phase of immune response and at the effector phase of antibacterial immunity. Cyclosporin A moderately decreased lysozyme activity and significantly decreased antibacterial activity peptides against Escherichia coli. Immunosuppressive effects of cyclosporin A were observed after immunoblotting with antibodies anti-G. mellonella lysozyme. Tricine SDS/PAGE shown that synthesis of antibacterial peptides of larvae treated with cyclosporin A was considerably inhibited. Insects of impaired immune response by cyclosporin A action lost protective immunity to insect bacterial pathogen P. aeruginosa.     

不同气候条件下慢性阻塞性肺疾病的病原学和免疫学研究

目的：探讨COPD患者不同气候条件下（雷暴、阴雨、台风）的病原学和免疫功能。方法：细菌鉴定采用VITEK全自动微生物鉴定系统。K．B纸片扩散法测定药敏。T淋巴细胞亚群测定采用单克隆抗体免疫组化荧光染色法。血清免疫球蛋白（IgG,IgA,IgM）用琼脂单向扩散法。结果：台风、雷暴、阴雨、天气气候条件60例患者痰培养阳性率分别为45％、40％、30％,占革兰阴性菌前2位分别为肺炎克雷伯杆菌、流感嗜血杆菌和肺炎克雷伯杆菌、大肠埃希杆菌及流感嗜血杆菌、铜绿假单胞菌。革兰阳性菌中分别以肺炎链球菌、金黄色葡萄球菌为主及肺炎链球菌、金黄色葡萄球菌为主和金黄色葡萄球菌、表皮葡萄球菌为主。铜绿假单胞菌、肺炎克雷伯杆菌、流感嗜血杆菌对常用抗生素具有较高的耐药性。台风、雷暴、阴雨气候条件下3组AECOPD患者CD3＋ CD4＋、CD4＋／CD8＋、IgG、IgA、IgM均明显低于健康对照组（P〈0．05）,CD8＋高于健康对照组（P〈0．05）,而发作期和缓解期无显著差异（P〉0．05）。三种气候条件下相互之间比较的细胞和体液免疫指标比较无显著差剐（P〉0．05）。结论：雷暴、阴雨、台风条件下AECOPD的患者存在病原学分布差异,对常用抗生素具有较高的耐药性,细胞和体液免疫功能进一步下降。但三者相互之间的免疫功能比较无明显差异。     

Characteristics of nonspecific resistance in children with lung diseases due to opportunistic microflora

Acute destructive pneumonia in children was found to be complicated by acute pleural empyema (APE) on days 3-21 of the disease. The time of the development of this complication depended on the state of the nonspecific resistance of the body: the greater was the degree of deficiency as manifested by cell-mediated and humoral immunity indices, the earlier developed APE. Staphylococci and Pseudomonas aeruginosa infected the pleural cavity of children under the conditions of essentially decreased phagocytic activity, phagocytic index, C'H50 and one of the classes of immunoglobulin. The reaction of the body to staphylococci and P. aeruginosa took its course after the type of primary or secondary immune response, depending on the time of infection.     

Lead and immunity: II. Suppression of humoral immune response to Hymenolepis nana in mice.

Serum protein changes in mice treated for varying durations with lead nitrate and subsequently infected with 1000 H. nana eggs were compared with their counterpart controls, only treated and only infected groups. Decreased values of beta and gamma globulins in all the experimental groups along with higher worm recoveries indicate suppression of humoral immune response by lead in association with higher worm recoveries indicate suppression of humoral immune response by lead in association with the cellular components since these globulins are known to contain antibodies. Lead treatment for a duration of 45 days proved to be most effective in suppressing the immune response.     

Pretreatment with the Gram-positive bacterial cell wall molecule peptidoglycan improves bacterial clearance and decreases inflammation and mortality in mice challenged with Pseudomonas aeruginosa

The objective of this study was to determine if inflammatory tolerance and enhancement of innate immune function could be induced by the Gram-positive cell wall component peptidoglycan (PGN). Male mice (C57BL6/J or C3H/HeJ, 8-12 weeks of age) were given intraperitoneal injections of 1mg PGN on 2 consecutive days. The mice were then challenged with lipopolysaccharide (LPS) or live Pseudomonas aeruginosa (1 x 10(8) colony-forming units) 2 days after the second pretreatment. Mice pretreated with PGN had diminished plasma concentrations of TNFalpha and IFNgamma and elevated concentrations of IL-10 in response to a subsequent LPS or Pseudomonas challenge when compared to untreated controls. Bacterial clearance was improved in mice pretreated with PGN, and mortality in response to a subsequent Pseudomonas challenge was significantly attenuated. PGN pretreatment of LPS-unresponsive mice (C3H/HeJ) verified that the effect of PGN pretreatment was not due to any LPS contamination. We have previously demonstrated that PGN pretreatment induced resistance to a Gram-positive bacterial challenge. The present study extends those results by showing that exposure to the Gram-positive bacterial cell wall component peptidoglycan also induces cross-tolerance to LPS and non-specifically enhances innate immune function in that PGN-pretreated mice had increased resistance to Gram-negative bacterial challenge.     

Delivery of CFTR by adenoviral vector to cystic fibrosis mouse lung in a model of chronic Pseudomonas aeruginosa lung infection

In cystic fibrosis (CF) there is an excessive inflammatory response to lung infections with Pseudomonas aeruginosa, which causes significant morbidity and mortality. Mice deficient in the cystic fibrosis conductance transmembrane regulator homolog (Cftr) have exaggerated production of proinflammatory cytokines in epithelial lining fluid and increased mortality in response to chronic bronchopulmonary infection with mucoid P. aeruginosa, compared with infected wild-type littermates. Whether delivery of CFTR to CF airways by an adenoviral vector (Ad2/CFTR-16) decreases cytokine production and mortality in response to chronic bronchopulmonary infection with mucoid P. aeruginosa was tested. CF mice [stock Cftrtm1Unc-TgN(FABPCFTR)#Jaw] were anesthetized with isoflurane and inoculated intranasally with either Ad2/CFTR-16, diluent (sucrose), or empty vector (Ad2/EV). Two weeks later, mice were anesthetized with 2.5% Avertin and inoculated transtracheally with P. aeruginosa-laden agarose beads (PA M57-15). The cumulative 10-day survival of mice pretreated with Ad2/CFTR-16 was significantly higher compared with mice pretreated with sucrose but not significantly higher than mice pretreated with Ad2/EV. After adjusting for differences in experiment, we found weight loss at 3 days for mice treated with Ad2/CFTR-16 to be significantly less than for the sucrose- or Ad2/EV-treated groups. However, cytokine responses were similar in all groups 3 days after infection. In conclusion, the observed survival advantage of adenoviral delivery of CFTR to the CF lung may be due either to CFTR expression or possibly to proinflammatory effects of the adenoviral vector, or both.     

The antibacterial immunity of people under dynamic observation in an altered radiation situation]

The comparative study of the isolation rate, level, antigenic and class specificity of serum antibodies to the causative agents of purulent septic infections (Staphylococcus aureus, Pseudomonas aeruginosa, Proteus mirabilis) and acute enteric infections in healthy adults with different ABO blood groups before (836 persons) and after (1,429 persons) the catastrophe at the Chernobyl nuclear power station was made. The study revealed the fact that the genesis of antibodies directed against different microorganisms can be stimulated without additional antigenic challenge in the form of disease or immunization, which was definitely indicative of the influence of small radiation doses in Kiev on the humoral immunity of the population. The multifactor character of the dependence of antibacterial antibody formation under altered radiation conditions on the specific features of the infective agent and the intensity of its circulation among the population, individual immune responsiveness of the body and concrete radiation conditions was established.     

铜绿假单胞菌的MALDI-TOF-MS检测方法的建立

目的 建立利用基质辅助激光解吸电离飞行时间质谱仪( MALDI-TOF-MS)对铜绿假单胞菌的快速检测方法.方法 通过MALDI-TOF-MS法对铜绿假单胞菌进行检测分析,并与生化鉴定方法相比较.结果 MALDI-TOF-MS对铜绿假单胞菌的检测后得到肽指纹图片及相关质谱数据,建立MALDI-TOF-MS对铜绿假单胞菌的快速检测方法.结论 MALDI-TOF-MS方法检测铜绿假单胞菌准确快速、操作简单等特点,可发展成为食品检验铜绿假单胞菌的重要(辅助)工具.     

Ablation of the complement C3a anaphylatoxin receptor causes enhanced killing of Pseudomonas aeruginosa in a mouse model of pneumonia

The C3a anaphylatoxin is a 77-amino acid peptide that is generated by enzymatic cleavage of C3 during activation of the complement system. C3a mediates numerous biological functions on binding its receptor (C3aR), which is present on both myeloid and nonmyeloid cells. To investigate the biological impact of C3a-mediated effects during acute pneumonia caused by Pseudomonas aeruginosa, we subjected C3aR-deficient mice and matched wild-type (WT) mice to P. aeruginosa pulmonary infection. C3aR-deficient mice exhibited increased killing of P. aeruginosa in the lungs, less dissemination of bacteria into the bloodstream, and a decreased inflammatory response to P. aeruginosa pulmonary infection compared with WT mice. To examine whether the absence of C3aR would impact the humoral immune response to P. aeruginosa, we immunized WT and C3aR-deficient mice via intraperitoneal injection with live P. aeruginosa. Both groups of mice developed similar levels of antibody specific to P. aeruginosa. Immunized C3aR-deficient and WT mice were subjected to P. aeruginosa pulmonary infection, and C3aR-deficient mice again displayed increased killing of P. aeruginosa in the lungs, less dissemination of bacteria into the bloodstream, and a decreased inflammatory response in the lungs. Collectively, these data demonstrate that independently of antibody production, absence of C3aR causes enhanced killing of P. aeruginosa despite a diminished inflammatory response in a mouse model of pneumonia.     

铜绿假单胞菌家兔肺部感染模型的建立与评价

目的:建立铜绿假单胞菌(PA,又名绿脓杆菌)肺部感染动物模型,并从病理学及细菌学、影像学等方面进行评价,研究铜绿假单胞菌对肺组织的致病作用。方法:24只健康清洁级新西兰大白兔随机分成2组,实验组采用经皮气管穿刺法,隔日注入铜绿假单胞菌反复感染家兔,对照组施以生理盐水。接种后隔日一次行胸部CT扫描。比较二组家兔的体温、血象及肺组织匀浆菌落计数;观察病理组织改变及胸部CT影像学变化。结果:注菌组家兔白细胞总数显著升高,肺组织匀浆菌落计数显著高于对照组。CT表现为双侧多发斑片状模糊影,部分可见实变及脓肿灶。大体标本可见家兔肺部结节、脓肿及出血灶,病理学镜下观察发现注菌组家兔肺组织内多个脓肿灶形成,大量中性粒细胞浸润,肺泡腔内大量渗出,部分出血,肺泡壁充血及肺水肿。随着注菌次数增多及时间推移,逐渐出现如肺泡间隔增厚、肉芽肿形成、实变等慢性肺部炎症的病理表现。结论:使用经皮气道穿刺法接种铜绿假单胞菌至新西兰兔肺部,可成功建立兔铜绿假单胞菌肺部感染模型。