New vaccine production platforms used in developing SARS-CoV-2 vaccine candidates

The threat of the current coronavirus disease pandemic, caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), is accelerating the development of potential vaccines. Candidate vaccines have been generated using existing technologies that have been applied for developing vaccines against other infectious diseases. Two new types of platforms, mRNA- and viral vector-based vaccines, have been gaining attention owing to the rapid advancement in their methodologies. In clinical trials, setting appropriate immunological endpoints plays a key role in evaluating the efficacy and safety of candidate vaccines. Updated information about immunological features from individuals who have or have not been exposed to SARS-CoV-2 continues to guide effective vaccine development strategies. This review highlights key strategies for generating candidate SARS-CoV-2 vaccines and considerations for vaccine development and clinical trials.     

Reactivity of human sera in a sensitive, high-throughput pseudovirus-based papillomavirus neutralization assay for HPV16 and HPV18

Sensitive high-throughput neutralization assays, based upon pseudoviruses carrying a secreted alkaline phosphatase (SEAP) reporter gene, were developed and validated for human papillomavirus (HPV)16, HPV18, and bovine papillomavirus 1 (BPV1). SEAP pseudoviruses were produced by transient transfection of codon-modified papillomavirus structural genes into an SV40 T antigen expressing line derived from 293 cells, yielding sufficient pseudovirus from one flask for thousands of titrations. In a 96-well plate format, in this initial characterization, the assay was reproducible and appears to be as sensitive as, but more specific than, a standard papillomavirus-like particle (VLP)-based enzyme-linked immunosorbent assay (ELISA). The neutralization assay detected type-specific HPV16 or HPV18 neutralizing antibodies (titers of 160-10240) in sera of the majority of a group of women infected with the corresponding HPV type, but not in virgin women. Sera from HPV16 VLP vaccinees had high anti-HPV16 neutralizing titers (mean: 45000; range: 5120-163840), but no anti-HPV18 neutralizing activity. The SEAP pseudovirus-based neutralization assay should be a practical method for quantifying potentially protective antibody responses in HPV natural history and prophylactic vaccine studies.     

云南省盈江县蚊虫调查及乙型脑炎病毒分离

1984年及1989年7～8月,在盈江县捕获成年雌性蚊虫7属32种632O只,霜背库蚊、三带喙库蚊和棕头库蚊是农村畜圈的主要蚊种,伪白纹伊蚊和白纹伊蚊是野外竹林区的优势蚊种。对所获蚊虫用C6/36细胞和小白鼠方法分离病毒,从三带喙库蚊和窄翅伊蚊中各分离到1株乙型脑炎病毒。分析认为三带喙库蚊是当地乙型脑淡病毒的主要传播媒介,窄翅伊蚊亦可参与该病毒的传播。     

自杀基因质粒表达载体pcDNA3.1(-)CMV.TK在鼻咽癌细胞中的构建及转染研究

目的 构建含TK自杀基因的质粒表达载体并进行转染研究。方法 根据已发表的Hsv-TK基因的核苷酸序列，设计并合成一对引物，以含TK基因的PGEM/TK质粒为模板扩增出TK基因全长CDS序列，将其克隆到质粒表达载体pcDNA3．1(-)CMV中，进行序列分析和酶切鉴定后，运用电穿孔法将重组体pCDNA3．1(-)CMV-CD车专人鼻咽癌CNE-2细胞中，观察其表达情况以及无毒前体药物GCV干预下对CNE-2细胞生长的抑制作用。结果 酶切和序列分析证明pcDNA3．1(-)(2MV．TK含完整的TK基因序列，RT-PCR从转染细胞总RNA中扩出预期片段；鼻咽癌CNE-2细胞在无毒前体药物GCV干预下，其生长受到抑制。结论 成功构建了质粒载体pcDNA3．1(-)CMVTK，可以将其作为鼻咽癌自杀基因治疗的一种载体。     

The potential health impacts of climate change: An overview

Climate change would have a range of impacts on human health. Health impacts would be caused by the direct effect of climatic factors on human health, such as heat stress, and possible changes in the frequency and intensity of extreme weather events such as storms, floods and droughts. Impacts on health would also be mediated by the indirect effects of climate change, such as changes in availability of food and water and the distribution of vector‐borne diseases. The majority of health impacts would be adverse and would depend greatly on the vulnerability of populations.     

A Comparison Study of Vector Velocity,Spectral Doppler and Magnetic Resonance of Blood Flow in the Common Carotid Artery

Magnetic resonance phase contrast angiography (MRA) is the gold standard for blood flow evaluation. Spectral Doppler ultrasound (SDU) is the first clinical choice, although the method is angle dependent. Vector flow imaging (VFI) is an angle-independent ultrasound method. The aim of the study was to compare VFI- and SDU-estimated peak systolic velocities (PSV) of the common carotid artery (CCA) with PSV obtained by MRA. Furthermore, intra- and inter-observer agreement was determined. MRA estimates were significantly different from SDU estimates (left CCA: p?<?0.001, right CCA: p?<?0.001), but not from VFI estimates (left CCA: p?=?0.28, right CCA: p?=?0.18). VFI measured lower PSV in both CCAs compared with SDU (p?<?0.001) with improved precision (VFI: left: 24%, right: 18%; SDU: left 38%, right: 23%). Intra- and inter-observer agreement was almost perfect for VFI and SDU (inter-observer correlation coefficient: VFI 0.88, SDU 0.91; intra-observer correlation coefficient: VFI 0.96, SDU 0.97). VFI is more accurate than SDU in evaluating PSV compared with MRA.     

Assessment of an immunomarking technique for the study of dispersal of Culicoides biting midges

Capture–mark–recapture techniques are used to determine the dispersal and survival of arthropods, including vector groups such as Culicoides. An assumption of these studies is that capture and the subsequent marking process does not impact of the survival and behaviour of the marked individual. The small size of Culicoides means that a significant mortality and disruption of normal behaviour such as host-location can be caused by the process of collection. Here we evaluate a technique, novel to the study of dispersal in vectors, to mark Culicoides directly and indirectly without prior capture. The acquisition and subsequent detection of marker protein by Culicoides exposed to a treated substrate was investigated in the laboratory. The technique was then assessed in a small-scale field trial where a defined section of resting habitat was sprayed with an egg white solution and Culicoides caught within the vicinity were tested for the presence of egg protein. It was found that up to 100% of Culicoides acquired the protein marker in the laboratory with no apparent impact on survival. In the field, pools of Culicoides obsoletus collected next to the treated area were found to be positive for the protein, suggesting that the technique could be used in larger-scale studies. The definition of a behaviourally non-invasive technique for marking Culicoides will greatly increase our understanding of the natural dispersal behaviour of Culicoides and other vectors.     

Immunization of bighorn sheep against Mannheimia haemolytica with a bovine herpesvirus 1-vectored vaccine

Mannheimia haemolytica is an important pathogen of pneumonia in bighorn sheep (BHS), consistently causing 100% mortality under experimental conditions. Leukotoxin is the critical virulence factor of M. haemolytica. In a ‘proof of concept’ study, a vaccine containing leukotoxin and surface antigens of M. haemolytica induced 100% protection in BHS, but required multiple booster doses. Vaccination of wildlife is difficult. BHS, however, can be vaccinated at the time of transplantation, but administration of booster doses is impossible. A vaccine that does not require booster doses, therefore, is ideal for vaccination of BHS. Herpesviruses are ideal vectors for development of such a vaccine because of their ability to undergo latency with subsequent reactivation which obviates the need for booster administration. The objective of this study was to evaluate the potential of bovine herpesvirus 1 (BHV-1) as a vector encoding M. haemolytica immunogens. As the first step towards this goal, the permissiveness of BHS for BHV-1 infection was determined. BHS inoculated with wild-type BHV-1 shed the virus following infection. The lytic phase of infection was superseded by latency, and treatment of latently-infected BHS with dexamethasone reactivated the virus. A recombinant BHV-1-vectored vaccine encoding a leukotoxin-neutralizing epitope and an immuno-dominant epitope of the outer membrane protein PlpE was developed by replacing the viral glycoprotein C gene with a leukotoxin-plpE chimeric gene. Four of six BHS vaccinated with the recombinant virus developed significant leukotoxin-neutralizing antibodies at day 21 post-vaccination, while two of six BHS developed significant surface antigen antibodies at day 17 post-vaccination. These antibodies, however, were inadequate for protection of BHS against M. haemolytica challenge. These data indicate that BHV-1 is a suitable vector for immunization of BHS, but additional experimentation with the chimeric insert is necessary for development of a more efficacious vaccine.     

Common Carotid Artery Flow Measured by 3-D Ultrasonic Vector Flow Imaging and Validated with Magnetic Resonance Imaging

Ultrasound (US) examination of the common carotid artery was compared with a through-plane magnetic resonance imaging (MRI) sequence to validate a recently proposed technique for 3-D US vector flow imaging. Data from the first volunteer examined were used as the training set, before volume flow and peak velocities were calculated for the remaining eight volunteers. Peak systolic velocities (PSVs) and volume flow obtained with 3-D US were, on average, 34% higher and 24% lower than those obtained with MRI, respectively. A high correlation was observed for PSV (r = 0.79), whereas a lower correlation was observed for volume flow (r = 0.43). The overall standard deviations were ±5.7% and ±5.7% for volume flow and PSV with 3-D US, compared with ±2.7% and ±3.2% for MRI. Finally, the data were re-processed with a change in the parameter settings for the echo-canceling filter to investigate its influence on overall performance. PSV was less affected by the re-processing, whereas the difference in volume flow between 3-D vector flow imaging and MRI was reduced to ?9%, and with an improved overall standard deviation of ±4.7%. The results illustrate the feasibility of using 3-D US for precise and angle-independent volume flow and PSV estimation in vivo.     

福州口岸诱卵器法的现场应用研究

目的进一步研究诱卵器法在口岸媒介监测现场的应用技术。方法诱卵器法。结果 （1）诱卵器放置不同高度时,0m组和1m组的诱卵指数分别为87.68和83.79,0m组高于1m组（χ2=13.24,P〈0.005）,0m组和1m组的幼虫密度分别为13.21条/个和11.23条/个,0m组高于1m组（t=2.259,P=0.05）;（2）诱卵器放置不同时间时,4d、7d和10 d组的诱卵指数分别为28.00、74.50和76.41,4 d组低于7 d组（χ2=86.54,P=0.000）和10 d组（χ2=92.69,P=0.000）,4d、7d和10d组幼虫密度分别为2.095条/个、9.11条/个和11.36条/个,4d组低于7d组（t=2.334,P=0.024）,7d组低于10 d组（t=4.691,P=0.000）;（3）诱卵指数和幼虫密度呈较为密切的正相关（0 m组r=0.779,P=0.008;1 m组r=0.794,P=0.006）,不同监测时间和监测点诱卵指数和容器指数的关系需进一步研究。结论在福州口岸建议诱卵器放置在地面上,放置时间为7～10d,以提高成蚊产卵率,增加敏感性;需进一步研究诱卵器法和布雷图指数、房屋指数和容器指数在口岸地区的关联性,探索评价指标的推广应用。