人类肠道病毒71型亚单位疫苗壳聚糖佐剂作用的研究

为了解人类肠道病毒71型(EV71)重组保护性衣壳蛋白(rVP1)亚单位疫苗壳聚糖佐剂的作用,本研究利用rVP1蛋白与壳聚糖佐剂制备的疫苗口服免疫新西兰白兔,通过ELISA方法检测兔血清、粘膜洗液(小肠、鼻腔和肺)及粪便中特异性IgG、IgA抗体水平,中和试验检测血清中和抗体滴度,并通过抗原刺激体外培养的兔脾淋巴细胞分泌细胞因子(IFN-γ、IL-4)的水平评价rVP1口服免疫效果。结果显示,单纯口服rVP1蛋白仅可诱导产生较低水平的血清IgG和粘膜IgA抗体,而含佐剂疫苗组与单纯抗原免疫组差异显著,并可诱导产生中和抗体。细胞免疫检测结果显示rVP1含佐剂组兔脾淋巴细胞分泌IFN-γ和IL-4水平明显高于未加佐剂组。本研究为研制EV71VP1口服疫苗奠定了基础。     

超抗原SEA增强小鼠对HBV DNA 疫苗的免疫反应

观察超抗原SEA(D227A)的真核表达载体(pmSEA),对HBVDNA疫苗诱导Balbc小鼠(H2d)免疫应答的调节作用。肌内注射空载体pcDNA3、HBVDNA疫苗加pmSEA佐剂(pHBVS2S+pmSEA)或不加佐剂(pHBVS2S);ELISA法测定血清抗HBs;ELISPOT检测分泌IFNγ的脾淋巴细胞;4h51Cr释放法检测小鼠脾细胞CTL活性。HBVDNA佐剂组免疫小鼠抗HBsAg抗体滴度明显高于不加佐剂组,其IgG1IgG2a的比例不同于多肽免疫组,二者分别为0.282与10。HBVDNA佐剂组均能增强IgG1和IgG2a的产生,是不加佐剂组的1.36、1.73倍。佐剂组小鼠脾淋巴细胞IFNγ的分泌量是不加佐剂组2～3倍。CTL细胞杀伤活性(E:T=100)佐剂组与不加佐剂组分别为:69.77%±7.5%、42.81%±7.7%,差异显著(P<0.05)。HBVDNA疫苗具有较强的免疫原性,能够诱导机体产生特异性的抗体及CTL反应;pmSEA佐剂能够提高小鼠对DNA疫苗的免疫应答,有望成为DNA疫苗的免疫佐剂。     

双歧杆菌处理体对弱抗原免疫应答的影响

目的：探讨双歧杆菌菌处理体在疫苗免疫中的载体／佐剂作用及对诱导免疫应答的影响。方法：将双歧杆菌菌处理体偶联乙肝病毒基因工程表面抗原(HBsAg)成拟菌颗粒免疫C57 BL／6小鼠,二次免疫后5w,ELISA方法测定小鼠血清抗-HBS的阳性率及抗体效价;MTT方法测定免疫小鼠NK的杀伤活性;RT—PCR方法测定免疫小鼠脾淋巴细胞中γ干扰素(IFN-γ)的表达;生物活性方法测定该小鼠腹腔巨噬细胞α干扰素(IFN—α)分泌,并与抗原对照组、商业乙肝疫苗组、完全福氏佐剂组(CFA)进行比较,评定菌处理体佐剂疫苗的效果。初步探讨菌处理体用于疫苗的机制。结果：菌处理体疫苗组抗-HBS的阳性率与效价、NK杀伤活性、IFN—α体内诱生水平与商业疫苗对照组比较差异有显著性(P＜0．05),与CFA比较差异无显著性(P＞0．05),在菌处理组及CFA组IFN—γ表达阳性,其他两组为阴性。结论：菌处理体佐剂具有明显增强弱抗原疫苗诱导免疫应答的作用。     

金线莲多糖对免疫抑制小鼠脾淋巴细胞体外增殖、分泌NO及细胞因子的影响

本试验旨在研究金线莲多糖(ARP)对免疫抑制小鼠脾淋巴细胞体外增殖、NO及细胞因子IL-2、IL-6、IFN-γ分泌水平的影响。MTT法检测小鼠脾淋巴细胞体外增殖;Griess法检测NO分泌水平;ELISA法检测细胞因子IL-2、IL-6、IFN-γ的含量。结果显示,与对照组比较,ARP在50~400μg/m L可明显促进免疫抑制小鼠脾淋巴细胞体外增殖(P0.01),促进NO分泌(P0.01),促进细胞因子IL-2、IL-6和IFN-γ分泌(P0.05,P0.01)。以上结果提示ARP能提高免疫抑制小鼠脾淋巴细胞体外免疫活性,其作用机制可能与促进免疫抑制小鼠脾淋巴细胞增殖,促进NO产生以及提高IL-2、IL-6、IFN-γ的分泌水平有关。     

双歧杆菌的完整肽聚糖对巨噬细胞产生IL-18的影响

目的探讨分叉双歧杆菌的完整肽聚糖对巨噬细胞功能的调节作用.方法以完整肽聚糖注射于小鼠腹腔,用ELISA法测定小鼠腹腔巨噬细胞产生的IL-18的含量.结果完整肽聚糖注射组小鼠腹腔巨噬细胞产生的IL-18的含量显著高于对照组(P＜0.01).结论分叉双歧杆菌的完整肽聚糖能激活巨噬细胞,并使之分泌多量的IL-18.     

不同佐剂肾综合征出血热纯化疫苗体液免疫效果的研究

在肾综合征出血热(HFRS)纯化疫苗原液中分别加入Al(OH)3、IL-2、GM-CSF、CFA等佐剂的一种或两种,以不加佐剂的纯化疫苗原液作为对照,分别免疫BALB/c小鼠,定期进行眼眶采血,用ELISA法检测血清中抗HFRS病毒的抗体水平。用不同佐剂的HFRS纯化疫苗免疫BALB/c小鼠后,其抗体产生的时间和滴度均不同。佐剂组与对照组相比抗体产生早且(或)滴度高,IL-2佐剂组在免后第3天就可以检测到抗体;联合使用两种佐剂组与单独使用一种佐剂组相比,抗体产生早、滴度高。结果表明,上述各种佐剂对HFRS纯化疫苗诱导BALB/c小鼠产生的免疫应答均有一定的增强作用;IL-2对早期免疫应答、早期抗体的产生有显著意义;联合应用两种佐剂疫苗的免疫效果优于只加其中一种佐剂的疫苗。     

SDF-1基因对HIV-1核酸疫苗诱导的免疫应答

研究SDF-1基因对HIV-1核酸疫苗诱导免疫应答的影响,以探求治疗性HIV-1核酸疫苗的新策略。将pCIneoGAG联合SDF1基因或者pCIneoGAG单独免疫Balb/c小鼠,采用ELISA检测免疫小鼠的特异性抗体和IFNγ水平,用MTT比色法检测免疫小鼠脾淋巴细胞的增殖,用乳酸脱氢酶(LDH)试验检测小鼠特异性细胞毒性T淋巴细胞(CTL)的应答。研究结果提示:与pCIneoGAG免疫组比较,pCIneoGAG联合SDF-1基因免疫组小鼠血清的抗HIV-1p24抗体滴度降低,有显著性差异(p<0.01);而与pCIneoGAG免疫组比较,pCIneoGAG联合SDF-1基因免疫组小鼠血清的IFNγ升高,差异显著(p<0.01);pCIneoGAG联合SDF-1基因免疫组小鼠的脾淋巴细胞增殖实验刺激指数(SI)以及特异性CTL活性均高于pCIneoGAG免疫组,有显著性差异(p<0.01)。因此,SDF-1基因联合HIV-1核酸疫苗免疫小鼠,可能增强特异性Th1细胞和CTL反应,SDF-1基因对体液免疫有抑制作用。SDF-1基因对于治疗性HIV-1核酸疫苗是具有较好应用前景的免疫佐剂。     

两种方法检测接种流行性出血热沙鼠肾细胞灭活疫苗后人体的中和抗体

本文应用空斑减少中和试验(PRNT)和细胞病变中和试验(cPENT)两种方法对出血热沙鼠肾细胞灭活疫苗扩大人体免疫后的血清进行中和抗体水平检测。根据两种方法对总计74人份的免疫后血清检测比较结果,两种方法检测的抗体阳转率和抗体水平(GMT)。CPENT法均高于PRNT法,经统计学处理均有显著性差异。不同免疫组的中和抗体水平比较结果,注射三针的阳转率(n=10,100％)高于两针组(n=10,20—30％);接种加氢氧化铝佐剂疫苗(n=13)较接种不加佐剂的两种疫苗(n=26)的抗体水平高,阳转率为92％—100％GMT为22—69;皮下途径(n=15)和肌肉途径(n=13)注射无明显差别,阳转率分别为87—93％和92—100％,GMT分别为29—46和22—61。以上结果进一步肯定沙鼠肾细胞疫苗的人体免疫性     

超抗原SEA增强小鼠对HBV DNA 疫苗的免疫反应

观察超抗原SEA(D227A)的真核表达载体（pmSEA）, 对HBV DNA 疫苗诱导Balb/c 小鼠(H2d）免疫应答的调节作用。 肌内注射空载体pcDNA3、HBV DNA 疫苗加pmSEA佐剂（pHBVS2S+pmSEA）或不加佐剂（pHBVS2S）; ELISA 法测定血清抗HBs; ELISPOT检测分泌IFN-γ的脾淋巴细胞;4 h51Cr 释放法检测小鼠脾细胞CTL 活性。HBV DNA佐剂组免疫小鼠抗HBsAg抗体滴度明显高于不加佐剂组,其IgG1/IgG2a的比例不同于多肽免疫组,二者分别为0.282与10。HBV DNA佐剂组均能增强IgG1和IgG2a的产生,是不加佐剂组的1.36、1.73倍。佐剂组小鼠脾淋巴细胞IFN-γ的分泌量是不加佐剂组2～3倍。CTL 细胞杀伤活性（E:T=100）佐剂组与不加佐剂组分别为：69.77%±7.5%、 42.81%±7.7%,差异显著(P<0.05)。HBV DNA 疫苗具有较强的免疫原性, 能够诱导机体产生特异性的抗体及CTL反应;pmSEA佐剂能够提高小鼠对DNA 疫苗的免疫应答,有望成为DNA 疫苗的免疫佐剂。     

新型结核病疫苗AEH/Al/IC在小鼠中的免疫原性研究

目的检测重组结核分枝杆菌(Mycobacterium tuberculosis,Mtb)融合蛋白Ag85B-ESAT6(Antigen 85 B,6-kDa early secretory antigenic target,AE)和热休克蛋白X(heat shock protein X,Hsp X)与氢氧化铝和聚肌胞苷酸(Polyinosinic-polycytidylic acid,poly I∶C)构建的新型结核病亚单位疫苗AEH/Al/IC在小鼠中的免疫原性。方法用疫苗(AEH/Al/IC)和佐剂(Al/Poly I∶C)分别免疫雌性BALB/c小鼠3剂次,每剂次间隔10 d。末次免疫后第10天,经ELISA检测小鼠血清中抗原特异性Ig G、Ig G1和Ig G2a的抗体效价,经酶联免疫斑点试验(enzyme-linked immunospot,ELISPOT)检测小鼠脾细胞分泌抗原特异性IFN-γ和IL-2的细胞频数,经ELISA检测小鼠脾细胞抗原特异性IFN-γ、IL-2和TNF-α的分泌量。结果与佐剂组相比,疫苗组诱导小鼠产生了高水平的AE和Hsp X特异性抗体;疫苗组诱导小鼠产生的AE和Hsp X特异性IFN-γ和IL-2阳性脾淋巴细胞(斑点形成细胞spot forming cells,SFC)均高于佐剂组,差异有统计学意义(IFN-γ:P0.05; IL-2:P0.05);疫苗组小鼠脾细胞AE特异性IFN-γ、IL-2和TNF-α分泌量明显较佐剂组高,差异均有统计学意义(P0.05),Hsp X特异性IFN-γ和TNF-α分泌量比佐剂组稍高,但差异均无统计学意义(P0.05)。结论结核病亚单位疫苗AEH/Al/IC具有良好的免疫原性,有希望成为一种新型的结核病预防候选疫苗。     

Recent advances in the study of Kaposi's sarcoma-associated herpesvirus replication and pathogenesis

It has now been over twenty years since a novel herpesviral genome was identified in Kaposi's sarcoma biopsies. Since then, the cumulative research effort by molecular biologists, virologists, clinicians, and epidemiologists alike has led to the extensive characterization of this tumor virus, Kaposi's sarcoma-associated herpesvirus(KSHV; also known as human herpesvirus 8(HHV-8)), and its associated diseases. Here we review the current knowledge of KSHV biology and pathogenesis, with a particular emphasis on new and exciting advances in the field of epigenetics. We also discuss the development and practicality of various cell culture and animal model systems to study KSHV replication and pathogenesis.     

The structure, reproduction and taxonomy of Vidalia and Osmundaria (Rhodophyta, Rhodomelaceae)

Comprises species occurring mostly in subtidal habitats in tropical, subtropical and warm-temperate areas of the world. An analysis of the type species, V. spiralis (Sonder) Lamouroux ex J. Agardh, a species from Australia, establishes basic characters for distinguishing species in the genus. These characters are (1) branching patterns of thalli, (2) flat blades that may be spiralled on their axis, (3) width of the blade, (4) primary or secondary derivation of sterile and fertile branchlets and (5) position of sterile and fertile branchlets on the thalli. Application of the latter two characters provides an important basic method for separation of species into three major groups. Osmundaria , a genus known only in southern Australia, was studied in relation to Vidalia , and its separation from the Vidalia assemblage is not accepted. Species of Vidalia therefore are transferred to the older genus name, Osmundaria. Two new species, Osmundaria papenfussii and Osmundaria oliveae are described from Natal. Confusion in the usage of the epithet, Vidalia fimbriala Brown ex Turner has been clarified, and Vidalia gregaria Falkenberg, described as an epiphyte on Osmundaria pro/ifera Lamouroux, is revealed to be young branches of the host, Osmundaria prolifera.     

New or rare chromosome counts in Artemisia L. (Asteraceae, Anthemideae) and related genera from Kazakhstan

Fifteen chromosome counts of six Artemisia taxa and one species of each of the genera Brachanthemum, Hippolytia, Kaschgaria, Lepidolopsis and Turaniphytum are reported from Kazakhstan. Three of them are new reports, two are not consistent with previous counts and the remainder are confirmations of very scarce (one to four) earlier records. All the populations studied have the same basic chromosome number, x = 9, with ploidy levels ranging from 2x to 6x. Some correlations between ploidy level, morphological characters and distribution are noted.     

肝癌中HBV和HCV基因和抗原的分布及意义

采用原位分子杂交方法检测HCV RNA及HBV X基因;采用免疫组织化学方法研究HCV核心抗原,非结构区C33c抗原及HBxAg在肝细胞肝癌中的定位及分布.结果表明(1)HCV RNA、HBV X基因在肝细胞肝癌组织检出率分别为40%(55/136)和82%(112/136).HCV RNA定位于癌细胞的胞浆内,阳性细胞呈散在、灶状及弥漫分布三种形式;HBV X基因在肝癌细胞中的分布呈胞浆型、核型及核浆型,阳性细胞也呈上述三种分布形式;(2)HCV C33c抗原、核心抗原在肝细胞肝癌中的阳性率为81%(133/164)及86%(141/164).C33c抗原定位于癌细胞及肝细胞的胞浆内;核心抗原既定位于癌细胞核中,又可定位于胞浆中.C33c抗原阳性细胞以灶状分布为主;而核心抗原阳性细     

Selection with two alleles and complete dominance

For a plant selection model with frequency-independent viabilities, fertilities and selfing rates, it is shown that apart from global fixation, for certain parameter combinations a protected polymorphism and facultative fixation (either allele may become fixed according to initial frequencies) may both occur. Facultative fixation requires different selling rates for the dominant and recessive type. Protection of the polymorphism requires resource allocation for male and female function. In this connection the problem of purely genetically caused population extinction is discussed.
For general frequency dependence and regular segregation, the chances for establishment of a completely recessive gene are compared to those of a completely dominant gene. It is proven that the process of establishment of the recessive gene, despite a fitness advantage, may be considerably endangered by drift effects if random mating prevails. The recessive gene may reach the same effectivity in establishment as a dominant gene, only if the recessive homozygote mates exclusively with its own type during the period of establishment.