风疹疫苗对人群免疫效应的研究

风疹是由病毒引起的急性出疹性呼吸道传染病,多见于儿童,成人也可发病。妇女在妊娠早期被感染,可通过胎盘引起胎儿感染,导致发生先天性风疹综合征(CRS),使胎儿缺损和先天性畸形,主要有先天性心脏病、白内障、耳聋和精神障碍等。接种风疹活疫苗是预防风疹的唯一有效措施。风     

HPV16 E6/E7重组DNA免疫对BALB/C小鼠细胞免疫的影响

目的：研究人乳头状瘤病毒16型(HPV16)E6／E7与结核杆菌热休克蛋白70(HSP70)重组DNA免疫对小鼠细胞免疫的影响。方法：采用肌肉注射DNA的方法免疫小鼠，用ELISA检测细胞免疫因子的表达水平，用RT-PCR的方法检测细胞中细胞免疫因子mRNA的水平。结果：以HPV16 E6／E7为基础的DNA免疫小鼠后，检测到的细胞免疫因子IL—2、IFNγ的含量明显升高；与结核杆茵HSP70重组后，IL—2、IFNγ的含量也较重组前明显升高。结论：以HPV16 E5／E7为基础的DNA免疫能增强小鼠的细胞免疫反应，与结核杆菌HSP70重组后更能提高DNA免疫的效果。     

DNA疫苗对感染结核分枝杆菌耐药株小鼠治疗效果的病理学评价

目的观察HSP65DNA、Ag85ADNA、Ag85ADNA联合利福平、Ag85A/ESAT-6嵌合DNA、Ag85A/ESAT-6嵌合DNA联合利福平对感染结核分枝杆菌耐药株小鼠治疗后的肺、肝、脾脏病理组织学变化,研究和评价该DNA疫苗治疗效果。方法将用结核分枝杆菌高耐利福平低耐异烟肼临床分离株HB361尾静脉注射17～19g的6～8周龄雌性BALB/c小鼠随机分为8组,每组10只。感染后3d开始,分别用生理盐水、pVAX1空载体、利福平、HSP65DNA、Ag85ADNA、Ag85ADNA联合利福平、Ag85A/ESAT-6嵌合DNA、Ag85A/ESAT-6嵌合DNA联合利福平治疗60d。治疗结束后3周,分别取肺、肝、脾脏组织观察病理改变,抗酸染色计数脾脏残余菌量。结果生理盐水组、pVAX1空载体组及利福平组小鼠肺、肝、脾脏组织病变严重,范围广泛,各DNA疫苗治疗组小鼠的肺、肝、脾脏组织病变得到不同程度改善,以Ag85ADNA、Ag85ADNA联合利福平组最为显著,该组小鼠脾脏组织中残余菌数也最少。结论根据小鼠肺、肝、脾脏组织病理学指标变化,能够客观且有效地评价DNA疫苗对结核小鼠的治疗效果,结果表明Ag85ADNA疫苗治疗结核分枝杆菌耐药株感染小鼠效果最优。     

不同佐剂在ESAT-6亚单位结核基因疫苗中的研究

目的 结核分支杆菌ESAT-6抗原是在人或动物结核病早期细胞介导免疫的主要作用靶点，本研究旨在评价不同佐剂在ESAT-6作为试验性结核疫苗方向的潜能。方法 将溴化二甲基双十八烷基铵（dimethyl dioctadecyl ammonium bromide，DDA）和单磷酰脂质体A（monophosphoryl lipid A，MPL）联合应用作为佐剂，用酶联免疫吸附试验（enzyme linked immunosorbent assay，ELISA）分析干扰素γ（IFN-γ）和免疫球蛋白G（IgG）含量，并比较2种佐剂的免疫效果。结果 将ESAT-6为基础的实验性疫苗与含有已知的保护性抗原成分的抗原85B复合体（antigen 85B，Ag85B）和短期培养滤液（ST-CF）等的疫苗进行平行比较，以DDA为佐剂免疫小鼠可以对Ag85B和ST-CF产生强的保护性免疫应答，而DDA佐剂用于ESAT-6时则作用轻微，未见细菌负荷的明显下降。采用DDA和MPL联合应用效果比单用DDA或单用MPL效果显著（P〈0．05）。结论 DDA和MPL的联合应用使得ESAT-6疫苗的接种成为可能，作为诊断性工具鉴别结核感染和接种免疫等方面具有强大潜能，为结核控制和预防提出了新的思路。     

从化市幼儿园学龄前儿童乙型肝炎疫苗免疫效果评价

<正>乙型肝炎是一种严重威胁人类健康的传染性疾病,据不完全统计,约有25%的乙肝携带者可以发展为肝硬化或者肝癌。为了解从化市学龄前儿童乙肝病毒的感染现状,为今后乙肝的防治工作提供指导性依据,我们总结分析了我市从1995年至2005年间共19 883例学龄前儿童的HBV检测资料,并将当年来我院体检的农村幼儿园学龄前儿童资料作为对照资料并进行比较。现将结果报道如下。     

不同免疫类型的柯萨奇病毒DNA疫苗免疫效果研究

柯萨奇病毒B组3型（CoxsackievimsgroupBtype3，CVB3）是引起人类病毒性心肌炎的主要病原体，目前还没有特异性的防治措施。我们将巨噬细胞源趋化因子（macrophage—derivedchemokine，MIC）基因和志贺毒素（Shigatoxin，STx）B亚单位基因分别与CVB3VP1基因融合构建DNA疫苗pcDNA3／MDC-VPI和pcDNA3／SYxB—VPI，免疫小鼠，用7LD30CVB3致死攻击，比较两种疫苗的免疫效果，为柯萨奇病毒疫苗的研制奠定试验基础。     

抗—HBs不同水平者对HB疫苗的免疫应答

广东省是乙型肝炎的高发区。近年来,对乙型肝炎疫苗(HB疫苗)的预防接种,已广泛开展。我们单位于1989年春,对HBV易感职工初次接种HB疫苗。3年后,义对抗-HBs不同水平的职工及幼儿园采取不同方案进行HB疫苗追加免疫或全程免疫,并通过血清学的观察,对HB疫苗应答状况进行了初探。材料和方法观察对象:本报社记者、编辑及其它工作人员,年龄为20～62岁。男215人,平均年龄为50岁;女101人,平均年龄为43岁;幼儿园儿童<5岁60名,     

狂犬疫苗免疫后的效果观察

被犬、猫等动物咬、抓伤后 ,绝大多数的人都能主动进行狂犬疫苗免疫预防。为了解注射狂犬疫苗后的抗体产生情况 ,对 1 0 85例前来我站全程注射狂犬疫苗者进行了血清学检测。现报告如下。材料与方法1 疫苗注射 :按说明书全程注射人用狂犬疫苗 (浓缩 )。疫苗均由省、市卫生防疫站逐级供应 ,有效期内使用。2 检测对象 :1 998年 1月至 1 999年 1 2月因被犬、猫等动物咬、抓伤来我站全程接种狂犬疫苗者 ,于最末一针两周后采血 ,当日检测。3 检测方法 :采用酶联免疫吸附试验(ＥＬＩＳＡ)。试剂盒系卫生部兰州生物制品研究所生产。有效期内使用 ,…     

增强核酸疫苗免疫应答的策略及方法

核酸疫苗又称DNA疫苗,是指将编码某种目的抗原蛋白的外源基因插入真核细胞表达质粒,直接导入动物体细胞内,并通过宿主细胞的表达系统合成抗原蛋白,诱导宿主产生对该抗原蛋白的免疫应答,从而达到预防和治疗疾病的目的.核酸疫苗不仅能刺激机体产生特异性的体液免疫,还能诱导产生特异性的具有细胞毒杀伤性功能的CTL效应（细胞免疫）,并可直接清除带有目的抗原的靶细胞,因此对病毒、细胞内寄生的细菌和寄生虫所引起的传染病具有广阔的治疗前景[1-6].     

长期多代饮奶近交BALB/C小鼠的骨代谢模型

背景:现有的研究结果大多是反映短期饮奶或单代饮奶对骨代谢的影响,而反映长期多代饮奶对骨代谢影响的研究较少.目的:观察长期多代饮奶对近交BALB/C小鼠骨代谢的影响.设计、时间及地点:完全随机设计,实验于2005-03/2006-03在郑州大学公共卫生学院实验室完成.材料:取4周龄近交BALB/C小鼠60只,雌雄各半,随机分为实验组和对照组,每组30只.红星牌全脂甜奶粉由黑龙江红星集团股份有限公司生产(批号20050412),每7.43 g加入13 mL烧开的去离子水,还原为浓缩4倍的牛奶.方法:实验组每日给予牛奶灌胃(0.083mL/g体质量),对照组每日给予相同剂量去离了水灌胃,连续灌胃6周.到小鼠发情期(约70 d左右)后,同组小鼠雌雄合笼交配,交配成功后将雌性小鼠分笼喂养,直到产下第2代.第2代小鼠到断奶期时(3周左右)断母乳,各组第2代继续分别用牛奶和去离子水灌胃,重复上述步骤得到第3代,断母乳后继续用牛奶和去离子水灌胃,第3代出生后17周左右实验结束.主要观察指标:每代在其子代出生后1个月,每组取20 只剪尾取血,采用全血干化学免疫浓缩法测定特异性骨碱性磷酸酶活性;3 d后再去眼球取血,离心取血清,采用邻甲酚肽络合酮比色法(终点法)测定血清钙浓度和血清碱性磷酸酶活性.采用磷钼酸比色法测定血清磷质量浓度.采用放射免疫分析法测定血清骨钙素质量浓度.结果:①血清钙、磷水平:实验组各代之间、对照组各代之间、各代中实验组和对照组之间,差异均无显著性意义(P>0.05).②血清碱性磷酸酶活性:第3代实验组碱性磷酸酶活性低于对照组(P<0.05).实验组第3代碱性磷酸酶活性低于前2代(P<0.05).③特异性骨碱性磷酸酶活性:第2代、第3代实验组特异性骨碱性磷酸酶活性低于对照组(P<0.05).实验组第3代的特异性骨碱性磷酸酶活性低于第1代(P<0.05).④血清骨钙素水平:在实验组3代之间有明显升高趋势,每两代之间差异均有显著性意义(P<0.05).第2代、第3代实验组骨钙素质量浓度高于对照组(P<0.05).结论:长期多代饮奶可增强BALB/C小鼠成骨细胞活性,促进骨骼生长发育.     

HBVDNA免疫的实验研究

从中国人血清中克隆出HBsAg adr亚型基因,采用pcDNA 3.1HisC真核细胞表达质粒进行基因重组。重组质粒pcDNA 3.1HisC-HBsAg分别用0.01、0．1、1、10和100μg腿部胫骨肌多点免疫BALB／C小鼠．4周时进行同样剂量加强免疫。初次免疫后4周和8周时分别检测HBsAg和HBsAb。结果表明,1μg以上剂量组加强免疫后．均可诱导小鼠产生特异性抗体。揭示HBV有发展DNA疫苗的可能性。     

Differential expression of an equivalent clonotype among BALB/c and C57BL/6 mice

The primary anti-phosphorylcholine (PC) response in BALB/c, C57BL/6, and congenic and recombinant inbred strains of these parental types has been examined in the splenic focus system. The frequencies of PC-specific precursors were shown to vary among these strains from 2 to 20 precursors per 10(6) splenic B cells. The distribution of these frequencies suggests that elements closely linked to or within the major histocompatibility complex may play a role in the determination of this parameter, although additional experiments are necessary to adequately assess this possibility. Moreover, all strains tested, regardless of immunoglobulin allotype, expressed monoclonal antibodies indistinguishable from the TEPC 15 myeloma protein (T15) clonotype. Further, the frequency of this clonotype in a given strain did not appear related to allotype, since both high and low T15 frequencies were found among strains of either the BALB/c (a(1)) or C57BL/6 (a(2)) allotype. The examination of normal serum for the T15 idiotype, however, revealed that only mice of the BALB/c allotype (a(1)) expressed the T15 idiotype in detectable quantities. After immunization with Diplococcus pneumoniae, sera from mice of the a(1) allotype consistently contained large quantities of the T15 idiotype, whereas sera from mice of the a(2) allotype exhibited various degrees of cross-reactivity with anti-T15 antibody. These results suggest that: (a) the allotype of an individual, although closely related to serum levels of an idiotype, is unrelated to the proportion of the precursor population which expresses that idiotype and; (b) the serum expression of a given idiotype may reflect regulatory processes, which act either during or before antigenic stimulation, rather than the actual clonotype representation in the repertoire. These findings indicate that distinctions must be made between the expression of idiotypic determinants within precursor B-cell populations and elements which regulate the subsequent appearance of those idiotypes in serum antibodies.     

维甲酸诱导BALB／C近交系小鼠产生腭裂的剂量研究

目的：研究维甲酸诱导BALB／C近交系小鼠产生腭裂所需的剂量。方法：将40只BALB／C近交系小鼠分成四组,每组10只,在妊娠10d时分别给予不同剂量的维甲酸,给药后观察所有孕鼠的反应,在妊娠15d时解剖部分孕鼠,取胎鼠头部,行冠状切片,苏木精一伊红染色法（HE染色）,进行观察。剩余孕鼠任其自然分娩,观察子鼠情况。结果：给予70mg／kg维甲酸后,孕鼠组绝大多数表现腭裂动物模型所需特征。结论：70mg／kg剂量维甲酸能够诱导BALB／c近交系小鼠产生可靠的腭裂动物模型。     

超声微泡靶向破坏介导HSV-TK/GCV抑制BALB/c-nu小鼠卵巢癌的实验研究

目的 探讨超声微泡靶向破坏介导下HSV-TK基因联合更昔洛韦对裸鼠卵巢癌的杀伤效应.方法 挑选成功种植卵巢癌细胞且状态良好的雌性BALB/c-nu小鼠40只,随机分为4组:A组,HSV-TK+超声微泡(MBs)+超声辐射(US)组;B组,HSV-TK+超声辐射(US)组;C组,HSV-TK组;D组,磷酸盐缓冲液(PBS)组.分别利用Western-Blot和real time RT-PCR检测各组瘤体组织内TK蛋白和mRNA的表达,TUNEL法检测不同组的肿瘤细胞凋亡,并比较各组之间的肿瘤生长状况及裸鼠生存时间.结果 A组TK蛋白、mRNA表达及肿瘤细胞凋亡率高于其他各组(P＜0.05),平均肿瘤体积A组显著低于其他各组(P＜0.05),但生存时间上A组同其他各组之间差异并无统计学意义.结论 超声微泡靶向破坏介导HSV-TK联合更昔洛韦可促进裸鼠卵巢癌细胞的凋亡,并抑制肿瘤生长速度,但短期观察并不能延长裸鼠生存时间.     

188Re-抗CEA单抗在结肠癌模型体内的生物学分布

目的　观察188Re抗CEA单抗 (C5 0 )在结肠癌生物体内的生物学分布 ,评估188Re -C5 0在机体内的导向作用。方法　将BALB/C小鼠结肠癌模型分组后行瘤体内局部注射 0 .5mCi188Re标记的C5 0 0 .16mg ,注药后 6h、12h、4 8h、96h分别进行显像 ,同时按组分批放血处死 ,测定肿瘤及正常组织的放射性分布。结果　注射标记抗体后 12小时 ,肿瘤区放射性浓聚最高 ,轮廓清晰。 12小时肿瘤区的放射性强度及T/NT值均最高 ,瘤 /肠高达 39.85± 2 1.36 ,瘤/肝 2 1.94± 12 .72 ,瘤 /血 16 .38± 9.6 9。 12小时后T/NT值下降 ,但至 4 8小时仍能保持在较高水平 ,如 4 8小时瘤 /肝12 .6 9± 10 .92 1、瘤 /血 5 .2 3± 7.3。结论　188Re -C5 0在生物体内能与肿瘤抗原发生特异性结合 ,具有很好的导向性 ,为其用于临床结肠癌导向诊断与治疗提供了可靠的依据。     

Astaxanthin-rich algal meal and vitamin C inhibit Helicobacter pylori infection in BALB/cA mice

Helicobacter pylori infection in humans is associated with chronic type B gastritis, peptic ulcer disease, and gastric carcinoma. A high intake of carotenoids and vitamin C has been proposed to prevent development of gastric malignancies. The aim of this study was to explore if the microalga Haematococcus pluvialis rich in the carotenoid astaxanthin and vitamin C can inhibit experimental H. pylori infection in a BALB/cA mouse model. Six-week-old BALB/cA mice were infected with the mouse-passaged H. pylori strain 119/95. At 2 weeks postinoculation mice were treated orally once daily for 10 days (i) with different doses of algal meal rich in astaxanthin (0.4, 2, and 4 g/kg of body weight, with the astaxanthin content at 10, 50, and 100 mg/kg, respectively), (ii) with a control meal (algal meal without astaxanthin, 4 g/kg), or (iii) with vitamin C (400 mg/kg). Five mice from each group were sacrificed 1 day after the cessation of treatment, and the other five animals were sacrificed 10 days after the cessation of treatment. Culture of H. pylori and determination of the inflammation score of the gastric mucosae were used to determine the outcome of the treatment. Mice treated with astaxanthin-rich algal meal or vitamin C showed significantly lower colonization levels and lower inflammation scores than those of untreated or control-meal-treated animals at 1 day and 10 days after the cessation of treatment. Lipid peroxidation was significantly decreased in mice treated with the astaxanthin-rich algal meal and vitamin C compared with that of animals not treated or treated with the control meal. Both astaxanthin-rich algal meal and vitamin C showed an inhibitory effect on H. pylori growth in vitro. In conclusion, antioxidants may be a new strategy for treating H. pylori infection in humans.     

Effect of MuLV-related genes on plasmacytomagenesis in BALB/c mice

The role of spreading somatic cell infections with ecotropic MuLV viruses in the induction of plasmacytomas in BALB/cAN pi mice was determined by constructing congenic mice that lacked the gene locus Cv that codes for ecotropic virus. DBA/2 mice that lack Cv on chromosome (chr) 5 carry a closely linked gene Rmcfr that determines resistance to infection with mink cell focus-forming viruses (MCF). Rmcfr was retrogressively back-crossed onto BALB/c for six successive generations to produce N6 mice. N6 mice were mated to each other to produce BALB/c.DBA/2 Rmvfr/Rmcfr homozygotes. This stock of mice lacked Cv, as demonstrated by DNA hybridization and were as fully susceptible to developing plasmacytomas as the parental BALB/c. A second congenic stock BALB/c.DBA/2 Rmcfr/Rmcfr Fv-1n/Fv-1n was also developed, but the mice of this stock showed a reduced incidence of plasmacytomas, as did BALB/c.DBA/2 Fv-1n/Fv-1n mice. These findings indicated Fv-1 or a gene closely linked to it conferred partial resistance to plasmacytomagenesis. In constructing the BALB/c.DBA/2 Fv-1n/Fv-1n stock, a "control" congenic BALB/c.DBA/2 Fv-1b/Fv-1b was also developed at N6. Surprisingly, this stock carried the Qa2+ trait. These mice were also partially resistant to plasmacytomagenesis, suggesting a gene on chromosome 17 (the location of Qa2) or a gene located elsewhere that regulates Qa2 expression is linked to a gene controlling partial resistance to plasmacytoma development.     

The diversity of the influenza-specific primary B-cell repertoire in BALB/c mice

The primary immune response of BALB/c mice to influenza (PR8) hemagglutinin (HA), a complex protein antigen, has been examined by the splenic focus assay, and the resulting monoclonal anti-HA antibodies have been characterized by their reactivity with heterologous viruses. The analysis of the primary B-cell response to HA revealed marked differences from responses previously defined for haptenic determinants. There were following differences: (a) the frequency of HA-specific B cells in both conventional and germ-free BALB/c mice was 1 in 1.0-1.5 X 10(5) splenic B cells, which is substantially lower than the frequency of B cells responsive to various simple haptenic determinants; (b) monoclonal anti-HA antibodies were predominantly of the IgA or IgM isotypes instead of IgG, which dominates antihapten responses; and (c) after immunization, the frequency of anti-HA-specific B cells increases by 10- to 50-fold, which is much greater increase than that observed after immunization with haptenic determinants. Fine specificity analysis of primary monoclonal HA-specific antibodies revealed extensive diversity and a considerable overlap with the specificities obtained from immune mice. Given the low overall frequency of HA-specific B cells, it could be calculated that the representation of most HA-specific clonotypes within the B-cell repertoire could not exceed 1 in 10(7) B cells. These findings indicate that the primary B-cell clonotype repertoire is extremely diverse and largely antigen independent in its generation.