Molecular histocompatibility beyond Tears: The next generation version

Human Leukocyte Antigens (HLA) matching at the serological level used to serve as the measure of histocompatibility between organ donors and recipients. With advancements in HLA typing methods more precise HLA mismatch assessment tools were developed to measure dissimilarities at the molecular level, collectively referred to as Molecular Mismatch load analysis tools. Currently, several software are aimed at deciphering the dissimilarities using somewhat different immunologic rationales. Our goal, in this review is to provide a basic overview of the different computational approaches, provide clinical cases to contextualize concerns regarding the lack of assessment of immunogenicity, and present our personal view regarding the gaps and the needs of our field.     

Immunogenicity and reactogenicity of high- vs. standard-dose trivalent inactivated influenza vaccine in healthcare workers: a pilot randomized controlled trial

Objectives

To compare immunogenicity, reactogenicity and acceptability of high- and standard-dose trivalent inactivated influenza vaccine (HDTIV, SDTIV) in 18- to 64-year-olds.

Efficacy,immunogenicity, and safety of a quadrivalent inactivated influenza vaccine in children aged 6–35 months: A multi-season randomised placebo-controlled trial in the Northern and Southern Hemispheres

Background

A quadrivalent split-virion inactivated influenza vaccine (VaxigripTetra?, Sanofi Pasteur; IIV4) containing two A strains (H1N1 and H3N2) and B strains from both lineages (Victoria and Yamagata) was approved in Europe in 2016 for individuals aged?≥?3?years. This study examined the efficacy and safety of IIV4 in children aged 6–35?months.

Antigenicity and immunogenicity of HIV-1 gp140 with different combinations of glycan mutation and V1/V2 region or V3 crown deletion

The carbohydrate moieties on HIV-1 envelope glycoprotein (Env) act as shields to mask conserved neutralizing epitopes, while the hyperimmunogenic variable regions are immunodominant in inducing non-neutralizing antibodies, representing the major challenge for using Env as a vaccine candidate to induce broadly neutralizing antibodies (bNAbs). In this study, we designed a series of HIV-1 gp140 constructs with the removal of N276/N463 glycans, deletion of the V1/V2 region and the V3 crown, alone or in combination. We first demonstrated that all the constructs had a comparable level of expression and were mainly expressed as trimers. Following purification of gp140s from mammalian cells, we measured their binding to bNAbs and non-NAbs in vitro and capability in inducing bNAbs in vivo. Antibody binding assay showed that removal of N276/N463 glycans together with the deletion of V1/V2 region enhanced the binding of gp140s to CD4-binding site-targeting bNAbs VRC01 and 3BNC117, and CD4-induced epitopes-targeting non-NAbs A32, 17b and F425 A1g8, whereas further deletion of V3 crown in the gp140 mutants demonstrated slightly compromised binding capability to these Abs. Immunogenicity study showed that the above mutations did not lead to the induction of a higher Env-specific IgG response via either DNA-DNA or DNA-protein prime-boost strategies in mice, while neutralization assay did not show an apparent difference between wild type and mutated gp140s. Taken together, our results indicate that removal of glycans at N276/N463 and deletion of the V1/V2 region can expose the CD4-binding site and CD4-induced epitopes, but such exposure alone appears incapable of enhancing the induction of bNAbs in mice, informing that additional modification or/and immunization strategies are needed. In addition, the strategies which we established for producing gp140 proteins and for analyzing the antigenicity and immunogenicity of gp140 provide useful means for further vaccine design and assessment.     

流行性出血热双价灭活疫苗的安全性和免疫原性观察

目的　观察流行性出血热 (EHF)沙鼠肾细胞双价灭活疫苗的安全性和免疫原性。方法　以高发疫区现场青壮年为对象 ,基础免疫 3针 ,一年后加强 1针。采用间接免疫荧光试验、微量细胞病变中和试验检测接种后荧光抗体和中和抗体 ,重点观察部分接种者免疫后 72h内的全身体温反应和局部副反应。结果　基础免疫后 2周 ,荧光抗体阳转率及几何平均滴度 (GMT)分别为99 .0 4%、2 4.5 1± 2 .0 6 ;中和抗体阳转率及平均滴度对 76 118(Ⅰ型 )为 91.30 %、18.2 7± 2 .2 1,对UR(Ⅱ型 )为 88.41%、12 .47± 2 .16。基础免疫后 1年 ,荧光抗体阳转率下降为 37.34% ,中和抗体总阳转率近 80 %。加强后 2周 ,荧光抗体和中和抗体阳转率均为 10 0 % ,中和抗体滴度对Ⅰ型为 37.0 9±2 .2 4,对Ⅱ型为 32 .6 1± 2 .0 5。接种后全身体温反应发生率为 0 .46 % ,局部反应发生率为 1.98% ,未见严重副反应发生。结论　流行性出血热双价灭活疫苗近期免疫效果良好 ,接种副反应率低     

肠病毒71型分离株的鉴定及免疫原性研究

目的 在对肠病毒71型(enterovirus 71,EV71)分离株进行鉴定的基础上,对其诱导小鼠产生的免疫应答水平进行研究,拟为进一步的EV71候选疫苗研究奠定基础.方法 超速离心纯化病毒后,采用磷钨酸负染法通过电镜观察病毒形态、大小;采用特异性EV71单抗通过间接免疫荧光法(IFA)检测病毒的特异性;合成EV71特异性引物,通过RT-PCR对病毒进行分子生物学鉴定;病毒的VP1基因序列测定后,与其他EV71序列进行比较,绘制种系发育树,确定病毒基因型;通过腹腔注射途径免疫小鼠,免疫后分离血清通过微量细胞病变抑制法测定血清中和抗体效价,ELISA法检测血清抗体水平和抗体亚型水平.结果 电镜下可观察到典型的圆形病毒颗粒,直径为20～30 nm,呈典型的肠病毒形态;085和087株病毒感染细胞中均能观察到黄绿色荧光,提示该两株病毒可与EV71单抗特异性结合;RT-PCR可以从病毒感染细胞中扩增出226 bp大小的特异性产物带,而采用CA16特异性引物则不能扩增出相应大小的产物带;基于VP1基因序列的种系发育分析显示,087和085株均为C4基因型;085和087株EV71免疫小鼠后可诱导产生能中和包括其本身在内的多个病毒株的中和抗体,针对同一株中和病毒,实验组间差异无统计学意义;针对不同中和病毒株,各实验组中和本株、523-07T株的能力明显高于FY-02T株(P<0.05);ELISA法检测结果显示,接种灭活前后的EV71病毒085和087株后均能诱导小鼠产生抗EV71特异性IgG;IgG亚型为IgG1/IgG2a混合型,灭活病毒免疫组小鼠血清EV71特异性IgG、IgG1、IgG2a水平明显高于活病毒免疫组(P<0.05).两株病毒之间差异无统计学意义.结论 本研究分离到的085和087株病毒均为EV71 C4基因亚型,并具有一定的免疫原性.     

DNA疫苗免疫原性的研究策略

DNA疫苗既能诱导体液免疫又能诱导细胞免疫,是近年来疫苗研究的热点领域,被称为疫苗史上的第三次革命.但DNA疫苗诱导特异性免疫应答的水平不高,限制了它的广泛应用.本文对提高DNA疫苗免疫原性的几种策略进行综述.     

鸡Tie-2受体胞外片段的原核表达及目的蛋白免疫原性的研究

目的研究鸡Tie-2受体胞外片段(配体结合域)的原核表达以及目的蛋白对小鼠的抗原性。方法用PCR方法从既往构建的包含鸡Tie-2受体胞外段基因的表达质粒扩增目的片段,进而构建PQE原核表达载体并诱导其目的蛋白的表达,将目的蛋白进一步纯化、复性并免疫小鼠,获得抗血清,用ELISA和Western blotting检测抗血清中抗体的存在。结果经酶切分析及测序鉴定表明获得了鸡Tie-2受体胞外目的片段的PQE原核表达载体,能高效表达目的蛋白,将其免疫小鼠可产生抗重组蛋白的抗体。结论用原核表达的方式可成功的获得鸡Tie-2受体胞外目的片段的蛋白,并对小鼠产生免疫原性。     

鼠TRAM蛋白B细胞表位预测及其免疫原性检测

目的 预测TRAM蛋白的二级结构和B细胞表位,为抗鼠TRAM单克隆抗体制备奠定基础。方法以TRAM蛋白的氨基酸序列为基础,采用Goldkeu计算机分析软件以及网络nnpredict二级结构分析软件对TRAM蛋白二级结构及B细胞表位预测。合成针对该表位的多肽,以此多肽为免疫原免疫兔,对其免疫原性进行检测。结果用多参数预测TRAM蛋白的二级结构和B细胞表位,综合评判表明：TRAM分子的第216～229位氨基酸满足亲水性、可及性和可塑性,在二级结构上位于蛋白伸展结构或无规则卷曲结构内,最可能为其优势B细胞表位。此多肽能诱导机体产生较高的抗体滴度,多克隆抗体具有高的特异性。结论TRAM分子的第216～229位氨基酸为其优势B细胞表位,这为制作B细胞优势短肽单克隆抗体提供了理论依据。     

微囊化猪胰岛的免疫原性研究

目的 经过体外实验,明确微囊化新生猪胰岛样细胞团（ＩＣＣ）的免疫原性。方法 通过胰岛－淋巴细胞混合培养（ＭＬＩＣ）后的淋巴细胞转化试验。以游离的新生猪ＩＣＣ和空微囊为对照,比较微囊ＩＣＣ组与对照组的淋巴细胞转化值。结果 微囊ＩＣＣ组淋巴细胞转化值显著低于游离ＩＣＣ组（Ｐ〈０．０５）,但明显高于空囊组（Ｐ〈０．０５）。结论 微囊包裹后的新生猪ＩＣＣ的免疫原性比游离的新生猪ＩＣＣ的免疫原性显著降低。提