A prime-boost concept using a T-cell epitope-driven DNA vaccine followed by a whole virus vaccine effectively protected pigs in the pandemic H1N1 pig challenge model

Influenza A virus (IAV) vaccines in pigs generally provide homosubtypic protection but fail to prevent heterologous infections. In this pilot study, the efficacy of an intradermal pDNA vaccine composed of conserved SLA class I and class II T cell epitopes (EPITOPE) against a homosubtypic challenge was compared to an intramuscular commercial inactivated whole virus vaccine (INACT) and a heterologous prime boost approach using both vaccines. Thirty-nine IAV-free, 3-week-old pigs were randomly assigned to one of five groups including NEG-CONTROL (unvaccinated, sham-challenged), INACT-INACT-IAV (vaccinated with FluSure XP® at 4 and 7 weeks, pH1N1 challenged), EPITOPE-INACT-IAV (vaccinated with PigMatrix EDV at 4 and FluSure XP® at 7 weeks, pH1N1 challenged), EPITOPE-EPITOPE-IAV (vaccinated with PigMatrix EDV at 4 and 7 weeks, pH1N1 challenged), and a POS-CONTROL group (unvaccinated, pH1N1 challenged). The challenge was done at 9 weeks of age and pigs were necropsied at day post challenge (dpc) 5. At the time of challenge, all INACT-INACT-IAV pigs, and by dpc 5 all EPITOPE-INACT-IAV pigs were IAV seropositive. IFNγ secreting cells, recognizing vaccine epitope-specific peptides and pH1N1 challenge virus were highest in the EPITOPE-INACT-IAV pigs at challenge. Macroscopic lung lesion scores were reduced in all EPITOPE-INACT-IAV pigs while INACT-INACT-IAV pigs exhibited a bimodal distribution of low and high scores akin to naïve challenged animals. No IAV antigen in lung tissues was detected at necropsy in the EPITOPE-INACT-IAV group, which was similar to naïve unchallenged pigs and different from all other challenged groups. Results suggest that the heterologous prime boost approach using an epitope-driven DNA vaccine followed by an inactivated vaccine was effective against a homosubtypic challenge, and further exploration of this vaccine approach as a practical control measure against heterosubtypic IAV infections is warranted.     

Strong cellular and humoral anti-HIV Env immune responses induced by a heterologous rhabdoviral prime-boost approach

Recombinant rhabdovirus vectors expressing human immunodeficiency virus (HIV) and/or simian immunodeficiency virus (SIV) proteins have been shown to induce strong immune responses in mice and rhesus macaques. However, the finding that such responses protect rhesus macaques from AIDS-like disease but not from infection indicates that further improvements for these vectors are needed. Here, we designed a prime-boost schedule consisting of a rabies virus (RV) vaccine strain and a recombinant vesicular stomatitis virus (VSV) both expressing HIV Envelope (Env). Mice were primed and boosted with the two vaccine vehicles by different routes and in different combinations. Mucosal and systemic humoral responses were assessed using enzyme linked immunosorbent assay (ELISA) while the cellular immune response was determined by an IFN-gamma ELISPOT assay. We found that an immunization combination of RV and VSV elicited the highest titers of anti-Env antibodies and the greatest amount of Env-specific IFN-gamma secreting cells pre- and post-challenge with a recombinant vaccinia virus expressing HIV(89.6) Env. Furthermore, intramuscular immunization did not induce antigen-specific mucosal antibodies while intranasal inoculation stimulated vector-specific IgA antibodies in vaginal washings and serum. Our results show that it is feasible to elicit robust cellular and humoral anti-HIV responses using two different live attenuated Rhabdovirus vectors to sequentially prime and boost.     

Multiple immunizations with adenovirus and MVA vectors improve CD8+ T cell functionality and mucosal homing

Recombinant adenovirus vectors and MVA vectors were used in prime boost vaccine regimens to address the impact of repeated immunizations on transgene product-specific CD8(+) T cell frequencies, phenotypes, function, and localization. We show that a regimen with three immunizations incorporating MVA, human adenovirus serotype 5 and chimpanzee-derived adenoviruses serotype 68 or 7 yields high transgene product-specific CD8(+) T cell frequencies in spleen, blood, lymph nodes, and peritoneal lavage. Furthermore, upon triple immunization increased frequencies of transgene-specific T cells were measured at mucosal sites such as mesenteric lymph nodes, intestinal epithelium, and Peyer's patches. Multiple dose vaccine regimens that markedly increase functionally active transgene-specific T cells and target them to the appropriate ports of entry may be important in protection against pathogens such as HIV-1.     

混合预充液吸入麻醉药液/气分配系数的推算值

目的 :测定晶体与胶体混合预充液的液 /气分配系数 ,比较液 /气分配系数的推算值与实测值。　　方法 :利用计算机编制程序 ,将乳酸林格液、琥珀酰明胶注射液 (佳乐施 )、库血和人体血浆 4种体外循环预充液排列组合后 ,随机抽取样本 10份 ,采用注射器 2次平衡法 ,利用气相色谱仪测定混合预充液在 37℃、33℃、2 9℃、2 5℃、2 1℃和17℃ 6个不同温度点的液 /气分配系数 ,并与推算值比较。　　结果 :混合预充液 3种吸入麻醉药的液 /气分配系数均与温度呈负相关 ,6个不同温度液 /气分配系数的推算值与实测值呈直线关系 (P<0 .0 5 ,地氟醚 r=0 .94,异氟烷 (异氟醚 ) r=0 .95 ,氟烷 r=0 .93)。　　结论 :由公式推算吸入麻醉药混合预充液的液 /气分配系数是可行的。     

Protective effect of isoorientin-2″-O-α-l-arabinopyranosyl isolated from Gypsophila elegans on alcohol induced hepatic fibrosis in rats

Alcohol abuse is one of the major causes of liver fibrosis, which shows a sharply increasing trend worldwide, yet effective therapeutic options for advanced alcohol fibrosis are limited. Recently we investigated the effect of anti-fibrosis by isoorientin-2&Prime;-O-α-l-arabinopyranosyl (IOA) isolated from Gypsophila elegans. During the experiment, the model group received alcohol only, and treatment groups received the corresponding drugs plus alcohol respectively, while the normal control group received an equal volume of saline. Analysis at the end of 24-week experiments showed that IOA could significantly improve the liver function, as indicated by decreasing levels of alanine transaminase, aspartate transaminase, alkaline phosphatase, γ-glutamyltransferase, interleukin-6 and tumor necrosis factor-α. Moreover, IOA could effectively inhibit collagen deposition and reduce the pathological tissue damage. Research on mechanism showed that IOA was able to markedly reduce lipid peroxidation, recruit the anti-oxidative defense system, and induce HSC apoptosis by down-regulating bcl-2 mRNA, as well as inhibit the expression of α-smooth muscle actin and transforming growth factor β1 proteins. In short, our results showed that IOA is effective in attenuating hepatic injury and fibrosis in the alcohol-induced rat model, which should be developed as a new drug to treat liver fibrosis and even cirrhosis.     

Electrophysiology reveals semantic priming at a short SOA irrespective of depth of prime processing

The otherwise robust behavioral semantic priming effect is reduced to the point of being absent when a letter search has to be performed on the prime word. As a result the automaticity of semantic activation has been called into question. It is unclear, however, in how far automatic processes are even measurable in the letter search priming paradigm as the prime task necessitates a long prime–probe stimulus-onset asynchrony (SOA). In a modified procedure, a short SOA can be realized by delaying the prime task response until after participants have made a lexical decision on the probe. While the absence of lexical decision priming has already been demonstrated in this design it seems premature to draw any definite conclusions from this purely behavioral result since event related potential (ERP) measures have been shown to be a more sensitive index of semantic activation. Using the modified paradigm we thus recorded ERP in addition to lexical decision times. Stimuli were presented at two different SOAs (240 ms vs. 840 ms) and participants performed either a grammatical discrimination (Experiment 1) or a letter search (Experiment 2) on the prime. Irrespective of prime task, the modulation of the N400, the ERP correlate of semantic activation, provided clear-cut evidence of semantic processing at the short SOA. Implications for theories of semantic activation as well as the constraints of the delayed prime task procedure are discussed.     

Genetics of Lesch's typology of alcoholism

It is widely accepted that dopamine and serotonin (5-HT) neurotransmission can be critically involved in the development of alcohol abuse and alcohol dependence. Lesch's typology of alcoholism has been gaining increasing popularity as it qualitatively differentiates patients into different treatment response subgroups. The aim of the present study was to evaluate a possible genetic background of Lesch's typology with special emphasis placed on dopamine- and serotonin-related genes. 122 alcoholics (the mean age: 35+/-9 years) were investigated. According to Lesch's typology, 58 patients were of type I, 36 patients of type II, 11 patients of type III, and 17 patients of type IV. Alcohol drinking and family history was assessed by means of a structured interview, based on the Semi-Structured Assessment for the Genetics of Alcoholism. 150 control subjects without psychiatric disorders were also recruited. The control group was ethnically-, age- and gender-matched to the patients. The DRD2 TaqIA, exon 8, and promoter -141C ins/del polymorphisms as well as COMT Val158Met, 5HTT 44 bp del in promoter, and DAT 40 bp VNTR polymorphisms were detected by means of PCR. No significant differences were observed when the whole group of alcoholics and the controls were compared. Similarly, there were no differences between either the Lesch type I or type II alcoholics and the control subjects. No significant differences were observed between type I and type II alcoholics. Alleles frequencies were not calculated for the Lesch type III and type IV alcoholics since the number of patients was too small. The present results argue against any major role of the investigated polymorphisms in either Lesch type I or type II alcoholism. More comprehensive studies are needed to define the role of the investigated polymorphisms in Lesch type III and type IV alcoholism.     

牙本质粘接剂直接盖髓的动物实验研究

目的：通过动物实验比较传统盖髓剂Ca（OH）：与2种牙本质粘接剂：Prime＆Bond NT和Gluma Comfort Bond直接盖髓的效果，观察牙髓对材料刺激的反应。方法：选用3条杂种犬共45颗牙进行盖髓实验，每条犬分为3组，5颗牙／组：第1组为Prime＆Bond NT，第2组为Gluma Comfort Bond，第3组采用Ca（OH）2对照。机械穿髓后，大量生理盐水冲洗，分别采用3种材料盖髓，玻璃离子充填。3只狗分别于7d、28d、70d处死，HE染色后光学显微镜观察。结果：7d：3组盖髓材料均有炎症反应；28d：2种粘接剂盖髓组均仍可以看到大量炎症细胞浸润，在穿髓孔附近牙髓坏死、消失。Ca（OH）2组也均有少量炎症细胞浸润；70d：2种粘接剂盖髓组均在穿髓孔附近出现牙髓坏死、消失。Ca（OH）2组在穿髓孔附近出现了钙化组织。结论：实验结果显示，牙本质粘接剂对牙髓具有较强的刺激性，不宜用于直接盖髓。