人甲型H7N9禽流感全病毒灭活疫苗和裂解疫苗在小鼠中的免疫原性研究

 目的   比较人甲型H7N9禽流感全病毒灭活疫苗和裂解疫苗在小鼠中的免疫原性,为该疫苗的类型选择提供初步依据。 方法   采用相同血凝素含量（5 μg）的H7N9全病毒灭活和裂解疫苗（含或不含氢氧化铝佐剂共4种类型）,分别对BALB/c小鼠进行1针或2针免疫。免疫后,用血凝抑制（hemagglutination inhibition,HI）试验检测血清抗体滴度,比较不同类型疫苗的免疫效果。 结果   小鼠免疫1针全病毒灭活疫苗后,全部血清阳转,HI抗体几何平均滴度（geometric mean titer,GMT）为149;免疫2针后,抗体GMT为243。小鼠免疫1针裂解疫苗后无抗体阳转;免疫2针后全部抗体阳转,GMT为139。两种疫苗添加铝佐剂后,诱导的HI抗体GMT仅略有增加。 结论   H7N9全病毒灭活疫苗在小鼠中的免疫原性较强。在同样类型和剂量的情况下,裂解疫苗需要免疫两次才能达到与全病毒疫苗相同的效果。铝佐剂对免疫原性提升不明显。     

亚单位流感疫苗免疫效果与人群血型相关研究

目的　探讨亚单位流感疫苗的免疫效果与 A、B、O血型的关系。方法　选择同一工作单位的 54名,年龄22~66岁之间(平均年龄27.38岁)健康人作为调查对象。于 2003 年 9 月接种亚单位型流感疫苗,应用血凝抑制试验(HI)进行了流感抗体检测,同时依据血型分类。结果　54 名被观察者均无局部或全身反应。H1N1、H3N2、B三个型别免疫阳转率分别达到96.40%~96 92%、93.62%~93.71%、90.41%~90.50%;抗体几何平均滴度分别增长5.83~5.88倍、4.63~4.70倍、6.26~6.32倍;H1N1型中 B型血组高于A型组差异有显著性意义( t=-0.2110P<0.05),其余各型别血型组之间均差别无显著性。结论　亚单位流感疫苗安全性好,免疫效果亦良好.提示:A、B、O不同种类血型对流感疫苗的免疫原性的反应存在一定差异。     

在6～18月龄儿童中评价流感减毒活疫苗的安全性、免疫原性及保护效果

本文报道用甲型流感病毒冷适应减毒株caA/川崎/9/86(H1N1)和caA/洛杉矶(H3N2)免疫儿童的结果,并对二价疫苗中毒株间的干扰作了初步探讨.将182名6～18月龄血清阴性的儿童以双盲法分为4组:H1N1组、H3N2组、H1N1及H3N2二价疫苗组和接种鸡胚尿囊液的对照组.试验用疫苗滴度均为10~(6.2)半数组织培养感染量(TCID_(50)).疫苗接种后密切观察1周:疫苗组儿童未见明显不良反应,轻微呼吸道症状及低热与对照组无显著性差异.鼻洗液的流感病毒分离率:免疫后6天H1N1组为16%,H3N2组为36%,二价疫苗组为55%;11天时H1N1组为0%,H3N2组为2%,二价疫苗组为8%.全部分离物经检验仍保留减毒特性而无毒力恢复迹象.疫苗接种后4周,H3N2组抗体阳转率达90%以上,抗体几何平均滴度(GMT)为7.2,均显著高于H1N1组.二价疫苗组的H3N2抗体阳转率与单价疫苗组无明显差别,但H1N1抗体仅为31%,明显低于单价疫苗组(83%).     

流行性感冒灭活疫苗临床研究

兰州生物制品研究所（简称兰州生研所）研制出的流行性感冒灭活疫苗（简称流感疫苗）根据国家关于《新药临床研究批件》要求进行Ⅰ Ⅱ期的临床研究。接种后进行一般反应观察,采用血球凝集抑制试验测定血清中3种抗体滴度,结果反应率为0．81％,HIN1,H3N2和B型抗体阳转率分别为96％,95．7％和98．7％,接种后的血清抗体GMT分别增长39．7％,33．2和45．2倍。该疫苗具有很好的安全性,人体接种后具有较好的免疫原性。     

酵母重组乙型肝炎疫苗对高危儿童的免疫原性

作者在违禁药品使用者聚集的地区挑选出38名乙型肝炎病毒感染标志阴性的易感儿童,按0、1、6月免疫方案,肌肉接种酵母重组乙型肝炎疫苗,每剂20μg.于接种前及接种后1、2、6、7和12个月采血,检测抗-HBs.结果表明:第1针接种后1个月,血清阳转率为6%,抗体几何平均滴度(GMT)为27mIU/ml;第2针接种后1个月,血清阳转率为80%,抗体GMT为75mIU/ml;第3     

MF59佐剂流感疫苗在老年人中的免疫原性和安全性

作者在大样本老年人群中 ,采用随机双盲法 ,连续三个流行季节对 MF5 9佐剂 (水包油乳剂 ) (FLUADTM组 )和无佐剂流感亚单位疫苗 (AGRIPPALTM组 )进行了对照研究。　　结果表明 ,MF5 9佐剂能明显增强疫苗的免疫原性。第 1次接种后 2 8天 ,FL UADTM组抗 B、A/H3N2和 A/H1 N1亚型的血凝抑制(HI)抗体几何平均滴度 (GMT)明显高于 A-GRIPPALTM组 ;FLUADTM组对 B、A/H3N2和 A/H1 N1亚型的 HI抗体 4倍以上增长率或者免疫后 2 8天 HI抗体滴度≥ 1 2 8的百分率明显高于 AGRIPPALTM组 ;免疫前后抗A/H3N2 (A/北京 /32 …     

抗H7N9流感病毒中和抗体快速检测方法的建立及应用

目的：对建立的抗H7N9流感病毒中和抗体快速检测方法进行方法学验证及初步应用。方法：分别采用不同代次细胞对高、中、低不同滴度的阳性血清进行多次平行检测,考察细胞代次对检验结果的影响;采用NIBSC提供的参考品对方法学的特异性进行验证;同时应用抗H7N9的阳性血清检测进一步评估方法的准确性和精密性;采用ELISA-MNT法和血凝抑制（hemagglutination inhibition,HI）试验分别接种H7N9灭活流感疫苗免疫的小鼠血清样本20份,分析两种方法检测结果的相关性。结果：采用ELISA-MNT中和法,使用不同代次MDCK细胞（25、30和35代）检测相同的血清样本的中和抗体滴度结果相同;该方法只对羊抗H7N9的血清具有较高保护力,对其他血清基本没有交叉反应;该方法准确性良好;该方法组间、组内精密性的平均变异系数分别为4%和11%。该方法测定H7N9型流感疫苗免疫后的小鼠血清抗体效价,其结果与HI抗体的相关系数为0.61,表明两种方法的检测结果之间呈良好的正相关性。结论：建立的微量病毒中和法能够满足H7N9流感病毒中和抗体效价检测的要求,可用于H7N9新型大流行流感疫苗的免疫评价。     

国产重组酵母乙肝疫苗对成人的免疫效果

目的:了解国产重组酵母乙肝疫苗(YDV)5μg和10μg对成人的安全性和免疫学效果.方法:199例研究对象随机分为2组,分别接受YDV 5μg和10μg,每人每月肌内注射1次,共3次(全程免疫)后1个月,进行免疫效果观察.检测血清乙肝表面抗体阳转率和乙肝表面抗体平均几何滴度(GMT),并观察2组的不良反应.结果:研究对象中未发现中、重度的局部和全身不良反应.接种5μg和10μg疫苗组的乙肝表面抗体阳转率分别为89.25%及98.11%;乙肝表面抗体GMT分别为18.88及54.47mIU·mL-1,2组的GMT差异有显著性(P＜0.05).结论:YDV成人免疫是安全的,10μg YDV抗乙肝表面抗体的GMT明显高于5μg YDV.     

补充益生菌对接种流行性感冒疫苗后抗体滴度影响的Meta分析

目的:研究补充益生菌对接种流行性感冒(以下简称流感)疫苗后抗体滴度的影响。方法:计算机检索PubMed,EMBASE,Cochrane Central Register of Controlled Trials,MEDLINE,CBM,CNKI,收集补充益生菌对接种流感疫苗后抗体滴度影响的研究,用Review manager 5.0软件进行Meta分析。采用均数和95%置信区间(95%CI)评价补充益生菌对接种流感疫苗后抗体滴度的影响。结果:纳入7项随机对照临床研究,共计476例患者,Meta分析结果表明:补充益生菌可提高接种流感疫苗后甲型H1N1型流感抗体滴度(MD=4.710,95%CI:0.53~8.89,P<0.00001;I²=97%);补充益生菌可提高接种流感疫苗后甲型H3N2型流感抗体滴度(MD=16.86,95%CI:0.87~32.85,P<0.0001,I²=100%);补充益生菌可提高接种流感疫苗后乙型流感抗体滴度(MD=3.04,95%CI:-0.77~6.85,P<0.0001,I²=96%)。结论:补充益生菌可提高接种流感疫苗后甲型H1N1流感、甲型H3N2流感和乙型流感抗体滴度。     

上呼吸道感染婴儿对麻疹-腮腺炎-风疹(MMR)疫苗的血清学应答

为了评价上呼吸道感染(URTI)对MMR疫苗免疫应答的影响,作者选出两组共235名12月龄婴儿进行试验.其中120名接种疫苗前7天无URTI症状和其他疾病症状,为无URTI组;115名有URTI症状至少3天,但不发热为URTI组.两组婴儿同时接种MMR联合疫苗,用相同方法于接种前和接种后6个月采集血样,用酶免疫分析(EIA)法检测血清中抗麻疹、腮腺炎、风疹抗体,若麻疹抗体滴度7200mIU/ml、腮腺炎抗体的EIA光密度值≥0.2、风疹抗体滴度≥10IU/ml坝为免疫有效.共采集到198名接种前后血样,其中URTI组97名,无URTI组101名.这198名婴儿在接种前均未检出抗麻疹、腮腺炎、风疹抗体.接种后183人(92.4%)有抗麻疹抗体,URTI和无URTI组的血清阳转率各为89.7%(87/97)和 95.0%(96/101).抗体几何平均滴度(GMT)各为1121.omIU/ml和1079.ZmIU/ml.180人有抗腮腺炎抗体,URTI和无URTI组的血清阳转率分别为92.6%(88/95)和 91.1%(92/101).抗体GMT分别为971.7和1019.4.免疫后无麻疹抗体的15名婴儿都产生腮腺炎抗体.198名婴儿均产生风疹抗体,其中197人抗体水平超过15IU/ml,URTI和无URTI组的风疹抗体GMT分别为89.4IU/ml和84.7IU/ml.     

季节性流感疫苗接种对大流行流感的影响研究

加拿大研究发现,季节性流感疫苗接种增加2009年大流行甲型流感(H1N1)感染的危险性,而澳大利亚研究未能证实这个发现.雪貂实验结果表明,以前的季节性流感感染能防御大流行甲型流感(H1N1),但以前的季节性流感疫苗接种则不能.模型研究显示,流感感染可导致对不同亚型的暂时性免疫.这些观察可以解释加拿大和澳大利亚的不一致发...     

Antibody response to tetravalent influenza subunit vaccine in patients infected with human immunodeficiency virus type 1

The capacity of patients infected with human immunodeficiency virus (HIV) to develop an adequate antibody response to influenza vaccination in relation to the CD4 cell count has been studied in a prospective study. A total of 73 subjects (54 HIV-infected patients and 19 healthy control persons) were vaccinated with influenza subunit vaccine containing 15 μg hemagglutinin of each of the following strains: A/Beijing/353/89(H3N2), A/Singapore/6/86(H1N1), B/Panama/45/90, and B/Beijing/1/87. Hemagglutinin inhibition (HI) antibody titers were determined prior to vaccination, 3 weeks afterwards, and at the end of the influenza season. The percentage of subjects with HI antibody titers above the assumed protective level was significantly lower in the HIV-infected patients for all 4 vaccine strains compared with those in the control group (7–26% and 42–74%, respectively). There was an association between CD4 cell count and antibody response to the B/Panama strain only. The serologic response to tetravalent subunit influenza vaccine is severely impaired in the majority of HIV-infected patients compared with control subjects. The results of this study challenges the recommendation to vaccinate HIV-infected patients.     

上海地区在校中学生接种甲型H1N1流感疫苗前后抗体水平监测分析

 目的  了解上海地区在校中学生接种甲型H1N1流感疫苗前后的抗体水平;观察甲型H1N1流感疫苗对该人群的免疫保护作用。 方法    应用常规微量血凝抑制试验（micro-hemagglutination inhibition test, HIT）对上海地区在校中学生分3个时间段进行甲型H1N1流感病毒抗体的血清学监测,3个时间段的抗体阳性率比较采用Pearson χ2 检验进行分析。 结果  2009年甲型H1N1流感流行前上海地区在校中学生的血清抗体阳性率仅为1.3%。经过一段时间流行后,血清抗体阳性率升至8.5%。接种甲型H1N1流感疫苗后,血清抗体阳性率升至87.3%。经Pearson χ² 检验分析,3个时间段在校中学生的血清甲型H1N1流感抗体阳性率差异有统计学意义（χ²=243.7,P<0.05）。 结论  在甲型H1N1流感流行前,上海地区在校中学生对其几乎没有免疫力,接种甲型H1N1流感疫苗能为该人群提供较好的免疫保护作用。     

Long-lasting protective immunity against H7N9 infection is induced by intramuscular or CpG-adjuvanted intranasal immunization with the split H7N9 vaccine

There is an urgent need for efficient vaccines against the highly pathogenic avian influenza A viral strain H7N9. The duration and intensity of the immune response to H7N9 critically impacts the epidemiology of influenza viral infection at the population level. However, the insufficient immunogenicity of H7N9 raises concerns about vaccine efficacy. In this study, we evaluated the impact of immunization routes and the adjuvant CpG on the immune response to a split H7N9 vaccine in mice. Determination of humoral and cellular responses to the vaccine revealed that after four vaccine doses, high titers of H7N9-specific serum IgG, determined by the influenza hemagglutination inhibition (HI) assay, were induced through the intramuscular (i.m.) route and lasted for at least 40 weeks. CpG-adjuvanted immunization increased the levels of long-lived IFN-γ⁺ T cells and raised the Th1-biased IgG2a/IgG1 response ratio. In addition, aside from mucosal IgA, CpG-adjuvanted intranasal (i.n.) immunization elicited serum IgG and cellular responses of a similar duration and intensity to CpG-adjuvanted i.m. immunization. Mouse challenge assays demonstrated that 24 weeks following i.m. immunization without CpG or CpG-adjuvanted immunization through the i.m. or i.n. routes, both offered a high level of protection against H7N9 infection. These results indicate that efficient long-term protection against H7N9 can be achieved via the optimization of vaccination strategies, such as immunization doses, routes, and adjuvants.     

四价流感病毒裂解疫苗生产用毒种传代稳定性研究

目的  研究四价流感病毒裂解疫苗生产用毒种的传代稳定性,确保毒种适用于疫苗生产。方法  将WHO推荐的季节性流感疫苗甲型H1N1和H3N2、乙型BV和BY病毒株（原始种子批）在鸡胚中传代扩增,制备生产用主种子批和工作种子批毒种。按中国药典2015年版三部的要求对毒种进行检定,包括鉴别试验、病毒滴度和血凝滴度测定、外源微生物检查等。将毒种连续传10代,用反转录PCR扩增第2、3、4、5、10代病毒鸡胚尿囊液中的血凝素（hemagglutinin,HA）和神经氨酸酶（neuraminidase,NA）基因片段,并进行序列测定,分析传代过程中HA和NA的核苷酸和氨基酸序列同源性。结果  主种子批和工作种子批毒种的抗原性均与推荐的病毒株一致。各型病毒滴度均>6.5 lg半数鸡胚感染量/ml,血凝滴度均≥1：160。2个种子批的支原体和无菌检查结果均为阴性,主种子批外源性禽白血病病毒和禽腺病毒检查结果亦为阴性。毒种在鸡胚中连续传代后,各型病毒株HA和NA的核苷酸序列同源性均>99%,氨基酸序列同源性均为100%。结论  本研究的四价流感病毒裂解疫苗生产用毒种具有良好的传代稳定性,可用于疫苗生产。     

The 2008-2009 H1N1 influenza virus exhibits reduced susceptibility to antibody inhibition: Implications for the prevalence of oseltamivir resistant variant viruses

A naturally-occurring H275Y oseltamivir resistant variant of influenza A (H1N1) virus emerged in 2007, subsequently becoming prevalent worldwide, via an undetermined mechanism. To understand the antigenic properties of the H275Y variant, oseltamivir resistant and susceptible strains of H1N1 viruses were analyzed by hemagglutination inhibition (HI) and microneutralization assays. HI analysis with H1-positive sera obtained from seasonal flu vaccine immunized and non-immunized individuals, and H1-specific monoclonal antibodies, revealed that resistant strains exhibited a reduced reactivity to these antisera and antibodies in the HI assay, as compared to susceptible strains. Neutralization assay testing demonstrated that oseltamivir resistant H1N1 strains are also less susceptible to antibody inhibition during infection. Mice inoculated with a resistant clinical isolate exhibit 4-fold lower virus-specific antibody titers than mice infected with a susceptible strain under the same conditions. Resistant and sensitive variants of 2009 pandemic H1N1 virus did not exhibit such differences. While HA1 and NA phylogenetic trees show that both oseltamivir resistant and susceptible strains belong to clade 2B, NA D354G and HA A189T substitutions were found exclusively, and universally, in oseltamivir resistant variants. Our results suggest that the reduced susceptibility to antibody inhibition and lesser in vivo immunogenicity of the oseltamivir resistant 2008-2009 H1N1 influenza A virus is conferred by coupled NA and HA mutations, and may contribute to the prevalence of this H1N1 variant.     

Characterization of a fully human monoclonal antibody against extracellular domain of matrix protein 2 of influenza A virus

The extra-cellular domain of the influenza virus matrix protein 2 (M2e) is highly conserved between influenza A virus strains compared to hemagglutinin and neuraminidase, and has long been viewed as a potential and universal vaccine target. M2e induces no or only weak and transient immune responses following infection, making it difficult to detect M2e-specific antibodies producing B-cells in human peripheral blood lymphocytes. Recently, using a single-cell manipulation method, immunospot array assay on a chip (ISAAC), we obtained an M2e-specific human antibody (Ab1-10) from the peripheral blood of a healthy volunteer. In this report, we have demonstrate that Ab1-10 reacted not only to seasonal influenza A viruses, but also to pandemic (H1N1) 2009 virus (2009 H1N1) and highly pathogenic avian influenza A virus, and that the antibody-bound M2e of 2009 H1N1 inactivated the virus with high affinity (∼10⁻¹⁰ M). More importantly, it inhibited 2009 H1N1 viral propagation in vitro. These results suggest that Ab1-10 might be a potential candidate for antibody therapeutics for a wide range of influenza A viruses.     

清新县甲型H1N1流感流行因素分析

[目的]通过系统监测调查,及时掌握我县甲型H1N1流感的疫情动态和流行规律,分析其流行因素,为我县完善甲型H1N1流感的防控工作提供科学依据.[方法]在清新县中部、南部、北部地区选择6个镇作为监测点,抽取普通人群静脉血,用酶联免疫试验检测甲型H1N1 IgG抗体进行血清学调查,同时以面对面问卷方式进行流行病学调查.[结果]共调查1325人,甲型H1N1流感抗体阳性190人,阳性率为14.34％;我县甲型H1N1流感病毒感染以轻症患者和隐性感染者为主;生活聚集性人群甲型H1N1IgG抗体高于非聚集人群(P＜0.05),甲型H1N1流感疫苗接种率为2.49％,免疫效果为57.58％;感冒患者就诊率为70.66％;甲型H1N1确诊0例.[结论]疫苗接种率及免疫效果、医疗诊断的准确率、生活聚集人群的易感性、以轻症感染和隐性感染为主的感染状况是影响我县甲型H1N1流感流行的主要因素,应加强人群甲型H1N1流感疫苗的接种,加强对临床医生甲型H1N1流感知识的培训,幼托机构、学校等聚集性场所仍然是我县甲型H1N1流感防控的重点场所.