禽用病毒-抗体复合物疫苗研究进展

病毒-抗体复合物疫苗是将病毒的特异性抗体按照适当的比例与传染性病毒混合制成的一种新型疫苗。国内外研究者做了大量的相关研究,并且在鸡传染性法氏囊病、新城疫等禽病的复合物疫苗研究中取得了突破性进展,为禽病疫苗的研究及禽病的防控开辟了一条新的独特途径。文章就复合物疫苗研究近况进行简要阐述。

基于病毒-抗体免疫博弈的WSN链路稳定算法

为解决超宽带无线传感器网络(WSN)存在的链路稳定性较差、数据传输遇阻等问题,提出一种基于病毒-抗体免疫博弈机制的超宽带WSN链路稳定算法.对节点所在区域进行均匀分割,通过设计覆盖划分方法并结合距离及剩余能量因素来优化分割区域,从而降低链路抖动概率.引入免疫算法,根据链路-节点之间的抗体特性来构建病毒-抗体免疫博弈机制,以优化节点及链路聚类效果,并通过病毒-抗体训练改善节点与链路间的数据交互特性,从而提升链路质量及区域传输性能.基于能量-跳数均衡方法,设计多参数判定机制,评估区域节点与sink节点的链路连通性能,提高算法的链路拥塞控制能力.在此基础上,利用PSK预发射方法的频域正交特性,对区域节点逐个设定发射频率,最大限度地降低因频率干涉而导致的链路抖动现象.仿真结果表明,与LEACH、LMS-A算法相比,该算法链路稳定性更高,网络稳定运行时间更长,拥塞发生频率更低.

病毒 - 抗体复合物疫苗应用研究进展

病毒－抗体复合物疫苗应用研究进展ＣｒａｉｇＥ．Ｗ．等著赵有编译傅先强校利用特异性高免血清或抗体按照适当的比例与传染性病毒混合做成病毒－抗体复合物，可作为疫苗（ＩＢＤＶ－Ａｂ）给动物接种。目前此项技术已成功地用于鸡传染性法氏囊炎的免疫。但对病毒和抗体的...

2012年江苏省健康儿童水痘-带状疱疹病毒抗体水平监测分析

目的了解江苏省健康儿童水痘—带状疱疹病毒抗体水平,为制定水痘的免疫学策略提供科学依据。方法采取随机抽样的方法抽取泰州市、常州市7岁以下健康儿童200名,采用酶联免疫吸附试验(ELISA)测定水痘-带状疱疹病毒免疫球蛋白G(IgG)抗体,用SPSS统计软件进行统计分析。结果共检测200名健康儿童的血清标本,水痘-带状疱疹病毒IgG抗体总阳性率为64.5%,抗体浓度均值为229.27 mIU/mL。不同年龄段健康儿童脊灰抗体阳性率有差异;有水痘疫苗免疫史的健康儿童的抗体阳性率及抗体平均浓度均明显高于无免疫史的儿童;不同地区健康儿童体内水痘-带状疱疹病毒IgG含量和抗体阳性率无显著性差异。结论接种水痘疫苗可获得较好的免疫保护。建议提高健康儿童水痘疫苗的覆盖率,同时改变水痘疫苗免疫策略,阻止疾病的传播流行。

面神经炎与抗水痘-带状疱疹病毒抗体的相关性探讨

目的 通过比较面神经炎患者与正常对照之间的抗水痘 带状疱疹病毒抗体滴度的差异以及面神经炎患者在不同疾病阶段的抗水痘 带状疱疹病毒抗体滴度变化 ,探讨抗水痘 带状疱疹病毒抗体与面神经炎之间的关联。方法 用酶标记葡萄球菌A蛋白 (SPA)染色法对 11例面神经炎患者及 2 6例正常人血清中抗水痘 带状疱疹病毒 (VZV)抗体进行检测 ,还对其中 6名患者进行了发病早期及 2周后的血清抗VZV抗体检测。结果 面神经炎患者组的抗VZV抗体滴度显著高于对照组 (P <0 .0 5 ) ;在 6例患者的不同疾病阶段测得的抗体滴度中 ,3例患者在发病 2周后抗体滴度比发病初期增加 4倍以上 ,1例增加 2倍 ,2例无改变。结论 抗VZV抗体滴度与面神经炎的发病相关 ,提示VZV感染可能是面神经炎的致病因素之一。

人血液和尿液样本HIV-1抗体的ELISA检测比较

目的 比较人的血液和尿液样本中HIV - 1抗体的一致性。方法 在广东省两所劳动教养所收集 346名HIV高危者的血液和尿液样本 ,分别用血液和尿液ELISA初筛试剂检测HIV - 1抗体。结果 346份血液和尿液样本各 18份阳性 ,其中有 17人的血液和尿液平行样本均为HIV - 1抗体阳性 ,血液和尿液样本HIV - 1抗体检测的一致性为 99.4 %。结论 在采集血液样本不便的情况下 。

中成药喜炎平雾化吸入治疗EB病毒抗体阳性鼻咽炎患者的临床观察

目的探讨中成药喜炎平雾化吸入治疗爱泼斯坦-巴尔(EB)病毒抗体阳性鼻咽炎患者的疗效。方法200例EB病毒抗体阳性鼻咽炎患者,采用简单随机方法分为治疗组和对照组,每组100例。对照组患者采用生理盐水进行雾化吸入治疗,观察组患者采用中成药喜炎平雾化吸入治疗。比较两组患者临床疗效及治疗前后鼻腔鼻窦结局测试-20(SNOT-20)量表评分。结果观察组患者的治疗总有效率为75.0%,显著高于对照组的25.0%,差异具有统计学意义(P<0.05)。治疗后,观察组患者的SNOT-20量表评分(10.56±0.57)分低于对照组的(21.17±0.21)分,差异具有统计学意义(P<0.05)。结论中成药喜炎平雾化吸入治疗EB病毒抗体阳性鼻咽炎患者疗效显著,值得临床推广应用。

Establishment of Monoclonal Antibody Competitive ELISA Using Monoclonal Antibody Against VP1 Protein of Asia 1 Type Foot-and-Mouth Disease Virus

Using the purified VP1 protein of Asia 1 type foot-and-mouth disease virus as the antigen, the purified monoclonal antibody was labeled by the sodium periodate method and the monoclonal antibody competitive ELISA was established in this study. Ten positive porcine foot-and-mouth disease serums and more than two hundreds negative serum were tested, and the results were the same as the background of samples. The sensitivity test and replicate test indicated that this method was stable and sensitive, which was suitable for monitoring Asia 1 type porcine foot-and-mouth disease virus antibody.

Comparison of Multiplex Fluorescent PCR with Serum Type-specific Antibody Detection in Diagnosis of Genital Herpes

Objectives: To compare multiplex fluorescent PCRwith serum type-specific antibody detection in thediagnosis of herpes simplex virus (HSV) infection andto evaluate its significance in the diagnosis of genitalherpes.Methods: We detected HSV infection in 121 speci-mens collected from patients with genital herpesusing both multiplex fluorescent PCR and serum type-specific antibody detection. HSV viral isolation wasused as the standard control.Results: When compared with the viral isolation, thesensitivity and specificity for multiplex fluorescentPCR were 100% and 88.89%, respectively afterdiscrepant analysis. The sensitivity and specificity fortype-specific antibody detection was 77.68 % and77.78 %, respectively. However, the type-specificantibody detected HSV in two asymptomatic patientswhile the multiplex fluorescent PCR couldn’t detectany HSV DNA from those specimens.Conclusions: Multiplex fluorescent PCR is a verysensitive and specific method for detection and typingof HSV in the lesion of genital herpes, it failed todetect HSV DNA from the asymptomatic patients.Serum type-specific antibody detection was a lesssensitive and specific test but could detect the specificantibody from some asymptomatic patients. Thecombination of these two techniques would allow rapid,sensitive and accurate detection and typing of HSVand help clinical diagnosis and epidemiologic survey-ing of genital herpes.

Application of high-titred IgY antibodies in orthopox virus diagnostics

Layer chickens were immunized with three species of inactivated orthopox virus （vaccinia virus, calpox virus and cowpox virus）. Antibodies （IgY） were purified from egg yolks by improved polyethylene glycol precipitation. The development of IgY directed against orthopox virus antigens was followed by immunofluorescence assay, plaque reduction neutraliztion test and immunoelectron microscopy. Cross-reactivity of two IgY antibodies with cells infected by the different strains of the pox viruses was also investigated by different methods （immunofluorescence assay, plaque reduction neutraliztion test and Western blot）. Even in very high dilutions in immunofluorescence assay （titres up to 1：10^6 and 1：10^5, respectively） and persisted on a plateau over 10 months after four booster injections, it was showed that anti-vaccinia virus IgY and anti-calpox virus IgY were positive. Neutralizing activity and ultra-structural detection of antigen with gold-labelled antibodies were respectively observed in plaque reduction neutralization test and immunoelectron microscopy. Western blot analysis revealed specific binding of IgY to virus proteins. Thus, there was cross-reactivity between different orthopox viruses. Finally, orthopox virns-specific IgY antibodies bounded magnetic beads （Dynabead） were used to concentration of orthopox viruses. This study suggests that anti-pox virus IgY could serve as a useful tool for orthopox viruses diagnosis.

Cost-effective method of siRNA preparation and its application to inhibit hepatitis B virus replication in HepG2 cells

AIM: To find a cost-effective method of preparation of short interfering RNAs based on cloning, fermentation, digestion and purification (CFDP) and test its feasibility to inhibit hepatitis B virus replication in cell culture. METHODS: We constructed an expression vector containing T7 and tac promoter in a head-to-head orientation. cDNA fragment of interest was cloned into this vector between the opposing promoters. dsRNAs were expressed with this vector in Escherichia coli, and purified by affinity chromatography using CF 11 column. They were digested by RNase III in a buffer containing manganese ions, then separated on 15% non-denaturing PAGE, and the siRNAs about 25 bp in length were recovered. siRNAs prepared with CFDP were co-transfected with target gene expression plasmid into human cell lines with lipofectamine 2 000 to test their inhibition efficiency. RESULTS: siRNAs corresponding to part of the hepatitis B virus polymerase gene (siHBVP) prepared by CFDP specifically and dramatically suppressed the virus protein expression. The HBsAg expression level was reduced to 10% that of the control by co-transfection of 60 nmol/L siHBVP in SMMC7721 cells. Dose-dependent effect on suppression of HBsAg and HBeAg expression was observed in HepG2 cells. The highest inhibition rate was kept at 70% during the six days after transfection of 7.5 nmol/L siHBVP. CONCLUSION: We show CFDP is a very promising method to prepare therapeutic agents in anti-virus applications.

Differential reactivity of mouse monoclonal anti-HBs antibodies with recombinant mutant HBs antigens

AIM: To investigate the reactivity of a panel of 8 mouse anti-hepatitis B surface antigen (HBsAg) monoclonal antibodies (mAbs) using a collection of 9 recombinant HBsAg mutants with a variety of amino acid substitutions mostly located within the “a” region.METHODS: The entire HBs genes previously cloned into a mammalian expression vector were transiently transfected into COS7 cells. Two standard unmutated sequences of the ayw and adw subtypes served as controls. Secreted mutant proteins were collected and measured by three commercial diagnostic immunoassays to assess transfection efficiency. Reactivity of anti-HBs mAbs with mutated HBsAgs was determined by sandwich enzyme-linked immunosorbent assay (ELISA).RESULTS: Reactivity of anti-HBs mAbs with mutated HBsAgs revealed different patterns. While three mutants reacted strongly with all mAbs, two mutants reacted weakly with only two mAbs and the remaining proteins displayed variable degrees of reactivity towards different mAbs. Accordingly, four groups of mAbs with different but overlapping reactivity patterns could be envisaged. One group consisting of two mAbs (37C5-S7 and 35C6-S11) was found to recognize stable linear epitopes conserved in all mutants. Mutations outside the “a” determinant at positions 120 (P→S), 123(T→N) and 161(M→T) were found to affect reactivity of these mAbs.CONCLUSION: Our findings could have important implications for biophysical studies, vaccination strategies and immunotherapy of hepatitis B virus (HBV) mutants.

A mimotope of Pre-S_2 region of surface antigen of viral hepatitis B screened by phage display

To acquire the phage-displayed mimotopes which mimic the specificity of hepatitis B virus surface antigen (HBsAg), a random peptide library expressing linear peptide with 12 amino asids in length were used to screen with the serum from a hepatitis B virus infected patient in the recovery phase. After 3 rounds of biopanning, the positive phages were confirmed by competitive ELISA using HBsAg/P33. Two phagotopes were identified and one of them was confirmed as mimotope by competition experiment. Based on the mimotpe, a multiple antigenic peptide with four branches was synthesized by solid phase peptide synthesis. The antiginicity and specificity of the synthesized antigen was tested in BALB/c mice compared with the native epitope-based antigen. The results showed that the mimotope-based antigen could evoke higher titer of antibodies with the same specificity of the epitope-based antigen. Those findings indicate mimotopes can be used in antigen and vaccine design.

Mutations in surface and polymerase gene of chronic hepatitis B patients with coexisting HBsAg and anti-HBs

AIM： To investigate the clinical significance and presence of mutations in the surface （S） and overlapping polymerase gene of hepatitis B patients with coexisting HBsAg and anti-HBs. METHODS： Twenty-three patients with chronic hepatitis B were studied. Of the 23 patients, i i were both positive for hepatitis B virus （HBV） surface antigen （HBsAg） and antibody to HBV surface antigen （anti-HBs）, 12 were negative for anti-HBs while positive for HBsAg. DNA was extracted from 200 μL serum of the patients. Nucleotide of the surface and overlapping polymerase gene from HBV-infected patients was amplified by PCR, and the PCR products were sequenced. RESULTS： Forty-one mutations were found within the surface gene protein of HBV in 15 patients （10 with coexisting HBsAg and anti-HBs）. Six （14.6%） out of 41 mutations were located at ＂α＂ determinant region in 5 patients （4 positive for HBsAg and anti-HBs）. Eleven mutations （26.8%） occurred in the downstream or upstream of ＂α＂ determinant region. Lamivudine （LMV）- selected mutations were found in three patients who developed anti-HBs, which occurred in amino acid positions （196, 198, 199） of the surface protein and in YMDD motif （M204I/V） of the polymerase protein simultaneously. Presence of these mutations did not relate to changes in ALT and HBV DNA levels.CONCLUSION： Besides mutations in the ＂α＂ determinant region, mutations at downstream or upstream of the ＂α＂ determinant region may contribute to the development of anti-HBs. These mutations do not block the replicating competency of HBV in the presence of high titer of anti-HBs.

Acute hepatitis B or exacerbation of chronic hepatitis B-that is the question

Hepatitis B virus (HBV) infection constitutes a serious global health problem. In countries with intermediate or high endemicity for HBV, exacerbations of chronic hepatitis B may be the first presentation of HBV infection. Some of these patients may be diagnosed mistakenly as having acute hepatitis B. Accurate diagnosis in these cases is very important for deciding whether to start treatment or not, because acute hepatitis B does not require therapy, while exacerbation of chronic hepatitis may benefit from it. Clinical and routine laboratory findings cannot help distinguishing between these two conditions. Therefore, several assays have been proposed for this purpose during the last few years. The presence of high levels of anti-HBe antibodies, HBsAg and HBV DNA are typical of chronic disease, whereas high titers of IgM anti-HBc, together with their high avidity index, characterize acute HBV infection. Starting from the description of a patient with acute hepatitis B-who recently came to our observation-we critically review the currently available assays that may help distinguishing between the different conditions and lead to the optimal management of each patient.

Squamous cell carcinoma of the anus-an opportunistic cancer in HIV-positive male homosexuals

Squamous cell carcinoma of the anus （SCCA） is a com- mon cancer in the human immunodeficiency virus （HIV）- infected population, and its incidence continues to in- crease in male homosexuals. Combined chemoradiation with mitomycin C and 5-fiuorouracil was poorly tolerated by severely immunocompromised patients in the early 1990s. In the era of highly active antiretroviral therapy （HAART）, however, recent data indicate that： （1） most HIV patients with anal cancer can tolerate standard chemotherapy regimens; and （2） this approach is associ- ated with survival rates similar to those of HIV-negative patients. However, HIV-positive patients with SCCA are much younger, more likely to develop local tumor recur- rence, and ultimately die from anal cancer than immune competent patients. Taken together, these findings suggest that anal cancer is an often fatal neoplasia in mid- dle-aged HIV-positive male homosexuals. In this popula- tion, SCCA is an opportunistic disease resulting in patients with suboptimal immune function from persistent infection and prolonged exposition to oncogenic human papillomaviruses （HPVs）. Large-scale cancer-prevention strategies （routine anuscopy and anal papanicolaou test- ing） should be implemented in this population. In addi- tion, definitive eradication of oncogenic HPVs within the anogenital mucosa of high-risk individuals might require a proactive approach with repeated vaccination.

Seroprevalence of anti-HAV among patients with chronic viral liver disease

AIM:To investigate the current seroprevalence of hepatitis A virus(HAV) antibodies in patients with chronic viral liver disease in Korea.We also tried to identify the factors affecting the prevalence of HAV antibodies. METHODS:We performed an analysis of the clinical records of 986 patients(mean age:49±9 years,714 males/272 females) with chronic hepatitis B virus(HBV) or hepatitis C virus(HCV) infection who had undergone HAV antibody testing between January 2008 and December 2009.RESULTS:The overall prevalence of IgG anti-HAV was 86.61%(854/986) in patients with chronic liver disease and was 88.13%(869/986) in age-and gendermatched patients from the Center for Health Promotion.The anti-HAV prevalence was 80.04%(405/506) in patients with chronic hepatitis B,86.96%(20/23) in patients with chronic hepatitis C,93.78%(422/450) in patients with HBV related liver cirrhosis,and 100%(7/7) in patients with HCV related liver cirrhosis.The anti-HAV prevalence according to the decade of age was as follows:20s(6.67%) ,30s(50.86%) ,40s(92.29%) ,50s(97.77%) ,and 60s(100%) .The antiHAV prevalence was significantly higher in patients older than 40 years compared with that in patients younger than 40 years of age.Multivariable analysis showed that age≥40 years,female gender and metropolitan cities as the place of residence were independent risk factors for IgG anti-HAV seropositivity. CONCLUSION:Most Korean patients with chronic liver disease and who are above 40 years of age have already been exposed to hepatitis A virus.

In vitro Anti-viral Activity of the Total Alkaloids from Tripterygium hypoglaucum against Herpes Simplex Virus Type 1

Herpes simplex virus type 1 （HSV-1） is a commonly occurring human pathogen worldwide. There is an urgent need to discover and develop new alternative agents for the management of HSV-1 infection. Tripterygium hypoglaucum （level） Hutch （Celastraceae） is a traditional Chinese medicine plant with many pharmacological activities such as anti-inflammation, anti-tumor and antifertility. The usual medicinal part is the roots which contain about a 1% yield of alkaloids. A crude total alkaloids extract was prepared from the roots of T. hypoglaucum amd its antiviral activity against HSV-1 in Vero cells was evaluated by cytopathic effect （CPE） assay, plaque reduction assay and by RT-PCR analysis. The alkaloids extract presented low cytotoxicity （CC50 = 46.6μg/mL） and potent CPE inhibition activity, the 50% inhibitory concentration （ICs0） was 6.5 μg/mL, noticeably lower than that of Acyclovir （15.4μg /mL）. Plaque formation was significantly reduced by the alkaloids extract at concentrations of 6.25 μg/mL to 12.5 μg/mL, the plaque reduction ratio reached 55% to 75% which was 35% higher than that of Acyclovir at the same concentration. RT-PCR analysis showed that, the transcription of two important delayed early genes UL30 and UL39, and a late gene US6 of HSV-1 genome all were suppressed by the alkaloids extract, the expression inhibiting efficacy compared to the control was 74.6% （UL30）, 70.9% （UL39） and 62.6% （US6） respectively at the working concentration of 12.5μg/mL. The above results suggest a potent anti-HSV-1 activity of the alkaloids extract in vitro.

Application of VP1 Protein to Develop Monoclonal Antibody against Foot-and-mouth Disease Virus Asia1 Type

In order to develop an anti-FMDV Asial type monoclonal antibody （mAb）, BABL/c mice were immunized with recombinant FMDV VP1 protein. Three mAbs, 1B8, 5El and 5E2, were then further optimized. The result indicated that prepared anti-FMDV Asial mAbs had no cross-reactivity with Swine vesicular disease （SVD） and FMDV O, A and C type antigen. Their titers in abdomen liquor were 1：5×10^6, 1：2×10^6 and 1：5×10^6, respectively. 1B8 was found to be of IgG1 subtype, 5El and 5E2 belonged to IgG2b subtype. In this study, the prepared mAbs are specific for detecting FMDV type Asia1, and is potentially useful for pen-side diagnosis.

Human Monoclonal Antibodies as Candidate Therapeutics Against Emerging Viruses and HIV-1

More than 40 monoclonal antibodies (mAbs) have been approved for a number of disease indications with only one of these (Synagis) - for a viral disease, and not for therapy but for prevention. However, in the last decade novel potent mAbs have been discovered and characterized with potential as therapeutics against viruses of major importance for public health and biosecurity including Hendra virus (HeV), Nipah virus (NiV), severe acute respiratory syndrome coronavirus (SARS-CoV), Ebola virus (EBOV), West Nile virus (WNV), influenza virus (IFV) and human immunodeficiency virus type 1 (HIV-1). Here, we review such mAbs with an emphasis on antibodies of human origin, and highlight recent results as well as technologies and mechanisms related to their potential as therapeutics.