HCV 多中和抗原表位嵌合病毒样颗粒免疫获得血清内中和抗体评价

目的:纯化的HCV 多中和抗原表位及HCV 包膜蛋白E2 嵌合的HBV S 抗原病毒样颗粒免疫新西兰兔,测定免疫血清内的中和抗体。分析中和抗体对HCVpp 感染Huh7.5 的中和作用。方法:纯化的HCV 多中和抗原表位及包膜蛋白E2 嵌合的HBV S 抗原病毒样颗粒(VLPs-MEpS、VLPs-E2S)10 滋g 皮下接种新西兰兔,间隔2 周共免疫3 次,采集不同时间免疫兔血清,ELISA 方法测定血清内的抗体,PBS 组作为对照;制备1b 型HCVpp,观察血清抗体对HCVpp 感染Huh7.-5 的中和作用,对免疫血清保护性进行初步评价。结果:免疫后血清中产生一定水平的中和抗体,血清中和抗体测定VLPs-MEpS 明显高于VLPs-E2S(P<0.05)。VLPs-MEpS 与VLPs-E2S 组均显著高于对照PBS 组(P<0.01)。对HCVpp 抑制作用,VLPs-MEpS 高于VLPs-E2S 组(P<0.05),最高中和率可达61.49%,二者均高于对照组(P<0.01)。结论:嵌合病毒样颗粒免疫新西兰兔后产生一定水平中和抗体,该中和抗体具有保护作用,为研发中和抗体表位疫苗奠定基础。     

Genetic studies on a mumps vaccine strain associated with meningitis

Vaccination with mumps measles and rubella (MMR) vaccine containing the live attenuated mumps strain, Urabe AM9, is associated with an increased incidence of meningitis. The isolation of mumps virus from CSF and subsequent identification as Urabe AM9-like by sequence analysis confirmed the causative role of Urabe AM9 vaccine in meningitis. To assess the role of genetic reversion in vaccine failure, sequence comparisons were made between several genes of Urabe AM9 vaccine and post-vaccination meningitis mumps isolates. An amino acid substitution in the Urabe AM9 HN gene Lys335Glu was not detected in the post-vaccination meningitis isolates suggesting that reversion to wild type sequence was associated with vaccine failure. However, further analysis showed that the vaccine was a mixture of viruses that differed at aa 335 of HN, possessing either the wild type Lys335 or the mutant Glu335, whereas the clinical isolates were homogeneous and possessed the wild type Lys335. Passage of the Urabe AM9 vaccine preparations in Vero cells resulted in the amplification of the Glu335 virus, however the post-vaccination meningitis isolates (Lys335) grew better in Vero cells than Urabe AM9 vaccine. A virus isolate, similar to the post-vaccination isolates was obtained from the vaccine suggesting that the strain responsible for vaccine failure was a pre-existing component of the vaccine and was not necessarily the result of reversion. The Urabe AM9 vaccine is a heterogeneous mixture of genotypes that differ in virulence with the HN Glu335 viruses being attenuated and at least a subset of the HN Lys335 viruses that are associated with disease. The Glu335 mutation may be among a class of attenuating mutations identified in several neurotropic viruses that involve charged amino acids in neutralising epitopes of receptor binding proteins. © 1998 John Wiley & Sons, Ltd.     

SCID-Hu mice immunized with a pneumococcal vaccine produce specific human antibodies and show increased resistance to infection.

Seventy-eight severe combined immunodeficiency (SCID) mice were administered intraperitoneally 1 x 10(7) to 9 x 10(7) human peripheral blood mononuclear cells (PBL) in five experiments. Human immunoglobulin G (IgG) was detected in 70 to 88% of these SCID-PBL-Hu mice after cell transplantation, and all four subclasses were present. The total concentration of human IgG varied from less than 1 to 10.2 g/liter. The SCID-PBL-Hu mice with high concentrations of human IgG regularly had mono- or oligoclonal human IgG bands in serum, as demonstrated by agarose gel electrophoresis. Of the SCID-PBL-Hu mice that were immunized with a 23-valent pneumococcal polysaccharide vaccine, 63 to 78% developed a significant human IgG antipneumococcal antibody response, whereas only very low levels of human IgM and no human IgA antipneumococcal antibodies could be detected. Twelve to twenty-two percent of the SCID-PBL-Hu mice showed signs of leakiness; these mice developed a significant mouse IgM antipneumococcal antibody response and no human antibodies. SCID-PBL-Hu mice were challenged intraperitoneally with 10 50% lethal doses of Streptococcus pneumoniae serotype 4 to study the protective effect of immunization with pneumococcal vaccine. The immunized SCID-PBL-Hu mice showed less bacteremia than did all control groups, and survival was 45 to 60%. None of the unimmunized SCID-PBL-Hu mice survived.     

Total serum IgE and specific IgE antibodies in children with bronchial asthma

Correlation between total serum IgE levels and RAST scores in a total of 342 asthmatic children were evaluated. The median and range of serum IgE were 1,050 IU/mL and 20 to 10,000 IU/mL respectively. Positive rates of RAST were highest for mites and house dust (87% to 91%) and lowest for milk, dogs, buckwheat, and eggs (2% to 8%). In general, patients with higher RAST scores had higher serum IgE (P less than .05, Mann-Whitney test). In individual cases, however, serum IgE levels did not significantly correlate with RAST scores and RAST was mandatory to estimate the levels of specific IgE antibodies.     

Ad5-HIVgag免疫的食蟹猴中腺病毒中和抗体与Gag特异性细胞免疫反应的研究

目的 观察不同剂量的重组腺病毒疫苗Ad5-HIVgag反复肌内注射免疫食蟹猴后,食蟹猴体内产生的腺病毒中和抗体水平及Gag特异性细胞免疫反应水平,初步探讨腺病毒中和抗体的产生对Gag特异性细胞免疫反应的影响.方法 将24只食蟹猴随机分为4组：对照组、低剂量组、中剂量组、高剂量组.每3周免疫1次,连续免疫5次.免疫前及免疫后不同时间点用中和实验检测动物体内腺病毒中和抗体水平,用Elispot方法检测Gag特异性细胞免疫反应水平.结果 初次免疫后3周3个实验组动物都检测到了较高水平的腺病毒中和抗体,在初次免疫后8周达到高峰,恢复期结束时略有下降.初次免疫后5周三个实验组动物都检测到了Gag特异性细胞免疫反应,除初次免疫后12周反应水平有所下降,其他时间点细胞反应呈逐渐增高趋势,但各剂量组间未见明显的量效关系.结论 在一定的剂量范围内反复多次应用Ad5-HIVgag免疫食蟹猴后,可产生针对腺病毒的中和抗体,随着免疫次数的增多Gag特异性细胞免疫反应有增高趋势,Ad5疫苗免疫后中和抗体的产生对Ad5疫苗反复应用诱导的Gag特异性细胞免疫反应抑制作用并不明显.     

Evaluation of a commercial assay for the detection of mumps specific IgM antibodies in oral fluid and serum specimens.

BACKGROUND: An IgM antibody capture radioimmunoassay (MACRIA) has been used in the UK Reference Laboratory for the detection of mumps specific IgM antibodies in oral fluid and serum samples. The method has proved both sensitive and specific and has been used for the surveillance of mumps since 1994. The use of radioactive labels has restricted the use of MACRIA to specialised laboratories and alternative, non-isotopic, sensitive assays capable of detecting the low levels of specific antibodies in oral fluid have not been available. Recently, a novel mumps specific IgM capture enzyme immunoassay (MACEIA) utilising recombinant mumps nucleoprotein (rMuVN) and monoclonal antibodies to the nucleoprotein, produced by Microimmune Limited, has been developed for use with both serum and oral fluid samples. OBJECTIVES: In this study, we have evaluated the performance of the Microimmune MACEIA for both serum and oral fluid against specimens tested by MACRIA. STUDY DESIGN: The panel consisted of matched serum and oral fluid specimens from 137 cases of suspected mumps received in the Virus Reference Department for routine investigation from March 2003 to October 2004. RESULTS: The sensitivity, specificity, positive and negative predictive value of the Microimmune MACEIA on serum samples compared to MACRIA were 98.8%, 100.0%, 100.0% and 98.5%, respectively. The sensitivity, specificity, positive and negative predictive value of the Microimmune MACEIA on oral fluid samples compared to Microimmune MACEIA and MACRIA consensus results on the paired serum samples were 90.3%, 97.6%, 98.3% and 87.1%, respectively. CONCLUSIONS: The Microimmune MACEIA was found to be a rapid, sensitive and specific alternative to MACRIA for the detection of mumps specific IgM in sera and oral fluids.     

流行性腮腺炎疫苗株S79全基因序列测定与分析

目的 腮腺炎疫苗株S79工作种子噬斑纯化及全基因组测序,针对国内已经分离出的毒株,从分子生物学角度探讨疫苗保护力相关问题.方法 噬斑纯化S79疫苗株,利用RT-PCR分段扩增S79株两种亚株的全基因并测序,比较S79株与其来源株Jeryl Lynn(JL)核酸序列差异;从GenBank中获得WHO各基因型参考株和中国腮腺炎流行株疏水蛋白(SH)序列,分析病毒株和疫苗株遗传距离,并构建基因亲缘性关系树.结果 S79疫苗株由两种亚株构成,比例为2∶5(major∶minor),获得全基因序列并递交GenBank;与相应的JL株相比,核苷酸序列同源性分别为99.7%和100%.亚株序列中存在散在区段异源重组.S79疫苗株为A基因型,中国的流行株为F基因型,遗传距离为11.2%～20.0%.结论 S79疫苗株由两种亚株构成,两亚株的全基因序列与JL株基本一致,但亚株的比例与国际上应用的其他JL疫苗株存在较大差异.疫苗株与中国流行株不属于同一基因型,遗传距离较远.S79疫苗免疫保护效果的差异可能主要与疫苗株和流行株不属于同一基因型有关.     

Virus excretion and strain specific antibody responses after oral poliovaccine in previously immunised children

A nation-wide campaign with trivalent oral poliovirus vaccine was organized in Finland in February-March 1985 in order to stop the unexpected outbreak of poliomyelitis. Excretion time of the vaccine viruses and antibody responses due to vaccination were studied in a group of healthy 6-year-old children who were classmates to one of the patients during the outbreak and who also had been screened for excretion of the epidemic poliovirus type 3 strain. While faecal excretion of at least one of the three vaccine virus serotypes was documented in all 19 children, only one throat specimen out of 106 studied was positive in the virus isolation test. The mean excretion times for types 1, 2, and 3 were 13, 21, and 21 days, respectively, and five children were still excreting a vaccine virus strain at 5 wk. Faecal excretion of the type 3 vaccine virus was not seen in children who had been excreting the epidemic type 3 strain 4 mo earlier. Excretion of a respective vaccine virus strain was usually well correlated to a booster response in serum neutralising antibodies to types 2 and 3 but not to type 1 poliovirus. A relatively high prevaccination antibody level did not always prevent the take of the corresponding vaccine virus strain. An increase in the level of neutralising serum antibodies towards at least one poliovirus serotype was observed in all but one of the 17 children studied. Antibody responses to the live vaccine strains were similar to those towards the corresponding nonattenuated strains while the absolute antibody titres against the epidemic P3/Finland/23127/84 strain remained relatively low in most sera studied.(ABSTRACT TRUNCATED AT 250 WORDS)     

Correlation of genetic variability with safety of mumps vaccine Urabe AM9 strain

The Urabe AM9 strain of mumps vaccine live is known for its genetic instability and some vaccines derived from this strain were withdrawn from the market due to an excessive number of vaccine-associated parotitis and meningitis cases. To identify the molecular basis of this instability, we determined complete nucleotide sequences of several stocks of the Urabe strain used for vaccine production by different manufacturers and of two clinical isolates from cases of vaccine-associated meningitis. In contrast to previously published studies relating the Lys335 --> Glu mutation in the viral HN gene with neurovirulence of mumps virus, we could not confirm any association of this mutation with the safety of mumps vaccine. Each of the three vaccine stocks studied had its own characteristic profile of mutations that was identified by cDNA sequencing and quantitated by mutant analysis by PCR and restriction enzyme cleavage. Determination of the mutational profile of mumps vaccine lots could allow vaccine manufacturers to characterize seed viruses and monitor the consistency of vaccine production to prevent emergence of virulent revertants.     

Characterization of neutralizing antibodies in adults after intranasal vaccination with an inactivated influenza vaccine

The levels and properties of neutralizing antibodies in nasal wash and serum collected from five healthy adults were examined after intranasal administration of an A/Uruguay/716/2007 (H3N2) split vaccine (45 μg hemagglutinin (HA) per dose; five doses, with an interval of 3 weeks between each dose). Prior to the assays, nasal wash samples were concentrated so that the total amount of antibodies was equivalent to about 1/10 of that found in the natural nasal mucus. Vaccination induced virus-specific neutralizing antibody responses, which increased with the number of vaccine doses given. Neutralizing antibodies were produced more efficiently in the nasal passages than in the serum: A ≥4-fold increase in nasal neutralization titres was observed after the second vaccination in four out of five subjects, whereas a rise in serum neutralization titres was observed only after the fifth vaccination. Nasal and serum neutralizing antibodies were mainly found in the polymeric IgA and monomeric IgG fractions, respectively, after gel filtration. Taken together, these results suggest that intranasal administration of an inactivated split vaccine induces high levels of nasal neutralizing antibodies (primarily polymeric IgA) and low levels of serum neutralizing antibodies (primarily monomeric IgG).     

双价痢疾菌苗角结膜免疫豚鼠后泪液中特异性抗体的检测

目的研究痢疾杆菌侵袭蛋白与局部免疫反应的关系,为痢疾菌苗的研制提供理论依据。方法用表达和不表达侵袭蛋白(Ipa+/Ipa-)的两株福氏、宋内氏双价痢疾菌苗经角结膜免疫豚鼠,并分别用福氏、宋内氏毒株攻击,BA-ELISA方法检测豚鼠泪液中特异性抗体升高情况。结果发现角结膜免疫后豚鼠泪液中特异性双价抗体水平较对照显著升高,差别具有统计学意义,Ipa+菌苗免疫组宋内氏抗体水平较Ipa-免疫组升高,差别具有统计学意义;攻击后,两免疫组局部抗体呈二次反应变化。结论侵袭蛋白的表达可加强痢疾菌苗诱导的局部抗体水平。两株双价痢疾菌苗均有较好的免疫原性。     

Persistence of immunity to varicella in children with leukemia immunized with live attenuated varicella vaccine

To determine the safety and efficacy of varicella vaccine, we studied 437 children in remission from leukemia who were immunized with live attenuated varicella virus. Three hundred one of the patients received two doses of vaccine and 136 received a single dose of vaccine from 1 of 10 lots from two manufacturers. The patients have been followed for an average of three years (range, one to six). Seroconversion occurred in 88 percent of the 437 children after the first dose of vaccine and in 98 percent after one or two doses. The proportions of patients who were seronegative after one, three, and five years were 20, 25, and 30 percent, respectively, with little change over time in the geometric mean titers of specific antibody (6.3, 6.5, and 5.7, respectively). Chickenpox has been documented in 36 vaccinated patients (8 percent) who had 3 to 640 vesicles (mean, 100), mild illness, and no complications. Of the 83 vaccinated patients exposed to varicella within their families, 11 had chickenpox; the attack rate was 14 percent (8 percent among seropositive patients, 29 percent among seronegative patients). There was no relation between the time since vaccination and either the attack rate or the severity of the breakthrough illness. Two doses of vaccine appeared to be no more effective than a single dose. Of the 372 patients receiving maintenance chemotherapy when immunized, 149 (40 percent) had a rash, which was treated with acyclovir in 16 children (4 percent) and became a severe febrile illness in 4. These reactions were not fatal and were all associated with vaccine lots, the use of which has since been discontinued. We conclude that in children in remission from leukemia, varicella vaccine is safe and induces an immunity to chickenpox that persists for more than three years.     

Functional state of specific immunity in children and teenagers vaccinated against mumps

The functional state of immunity was evaluated from the avidity index (AI) of specific antibodies (IgG) and the level and spectrum of their neutralizing activity. The study recruited 200 subjects immunized with Russian vaccine against mumps according to the mandatory scheme. A group of vaccinees with a low AI of specific IgG was identified mainly among old children and teenagers. The vaccinees with a low AI had a significantly lower protective immunity (as shown from the level and spectrum of serum neutralizing activity) than those with a high AI. The vacinees with no humoral, incomplete, or complete postvaccination immunity, but with a low AI of specific IgG, can constitute a population stratum that preserves sensitivity to wild-type mumps viruses and serves as a favorable medium for their circulation.     

A virological description of serous meningitis in children immunized with vaccine against epidemic parotitis

The morbidity structure was analyzed in children vaccinated against epidemic parotitis in 1993-2002. Eight children (4 with serous meningitis and 4 with lesions of the salivary glands) underwent virologic and immunologic examinations. The molecular typing of the SH-gene fragment of the parotitis virus showed the process in 7 cases to be provoked by the vaccination strain. Presumedly, progressing vaccine-associated meningitis inhibits antibody formation. The total incidence of vaccine-associated meningitis was shown, according to Saint Petersburg data, to be not high, which testifies to a low reactogenicity of the Russian vaccine strain.     

Enzyme immunoassay of mumps virus in cell culture with peroxidase-labelled virus specific monoclonal antibodies and its application for determination of antibodies

Mumps neutralizing monoclonal antibodies (MAs) were purified and labelled with horseradish peroxidase and used to detect virus-infected Vero cells, which were seeded as monolayers in wells of 96-well plates. This direct enzyme immunoassay (EIA) in cell culture proved to be a sensitive method for detection and titration of mumps virus and it may be useful for diagnostic purposes. The EIA is also suitable for the rapid determination of neutralizing antibodies. Neutralization of mumps virus by preincubation with either monoclonal or polyclonal antibodies was indicated by inhibition of the absorbance at 450 nm as measured with a multichannelled photometer. The EIA (duration 2 days) for determination of mumps neutralizing antibodies is an attractive alternative for the plaque reduction test (duration 6 days).     

Detection of antibodies to pre-early nuclear antigen and immediate-early antigens in patients immunized with cytomegalovirus vaccine.

To define the role of antibody to immediate early antigens of human cytomegalovirus (CMV) in diagnosing acute infection, we studied antibody responses of adult volunteers and renal transplant candidates receiving CMV vaccine. In addition, CMV antibodies in healthy adults were determined. Antibody to pre-early nuclear antigen, one of the immediate early antigens, did not develop in all of the volunteers or renal transplant candidates receiving the vaccine. In those vaccinees who developed an antibody response, the duration of the response was variable. Antibody to pre-early nuclear antigen and immediate early antigens was also detected in some healthy adults with serological evidence of past CMV infection, but without clinical evidence of recent infection. In view of these results, antibody to immediate early antigens and specifically to pre-early nuclear antigen does not appear to be suitable for rapid diagnosis of acute CMV infection.