Immunogenicity when utilizing adenovirus serotype 4 and 5 vaccines expressing circumsporozoite protein in naive and Ad5 immune mice

ABSTRACT: BACKGROUND: Induction of potent long lasting effector T cell responses against liver stage malaria antigens strongly correlates with protection from malaria. While Adenovirus serotype 5 (Ad5) based malaria vaccine platforms have the ability to induce potent effector T cell responses against transgenes, high rates of pre-existing Ad5 immunity in malaria endemic regions has prompted study of alternative Ad serotype based malaria vaccines as replacements for Ad5 based malaria vaccines. The research described in this article examines the utility of alternative serotype adenovirus serotype 4 (Ad4) expressing a sporozoite surface protein (circumsporozoite protein (CSP)) (Ad4-CSP) to induce immune responses against CSP. The immunogenicity of Ad4-CSP was also tested in homologous and heterologous prime boost vaccinations in both Ad5 naive and Ad5 immune backgrounds as compared to use of Ad5-CSP. RESULTS: In Ad5 naive animals, use of Ad4-CSP priming vaccinations followed by boosting with Ad5-CSP (Ad4-CSP/Ad5-CSP) maximally increased the numbers of CSP specific cytokine secreting cytotoxic T cells relative to repeated use of Ad5-CSP. The Ad4-CSP/Ad5-CSP regimen also induced equivalent levels of CSP specific cell killing as did homologous prime-boost vaccinations with Ad5-CSP, despite stimulating lower numbers of CSP specific cytotoxic T cells. Priming with Ad4-CSP followed by a homologous boost resulted in significantly less CSP specific humoral responses than any other vaccination regimen tested in Ad naive animals. In Ad5 immune animals, addition of Ad4-CSP in homologous or heterologous prime boost resulted in inductions of higher CSP specific responses than animals repeatedly vaccinated with Ad5-CSP alone. However, the observed responses were well below those observed in similarly treated Ad naive mice. CONCLUSIONS: While the Ad4-CSP/Ad5-CSP and Ad5-CSP/Ad5-CSP vaccination regimens resulted in equivalent CSP specific killing in Ad naive animals, Ad4-CSP/Ad5-CSP achieved this result with a lower percentage of CSP specific CD8+ T cells and a higher number of IFNgamma secreting cells, suggesting that the Ad4-CSP/Ad5-CSP vaccination regimen elicits more efficient cytotoxic T cells. In Ad5 immune animals use of Ad4-CSP improved CSP specific immune responses as compared to repeated use of Ad5-CSP, but could not achieve the levels of immunogenicity observed when the same vaccine regimens were used in Ad naive animals. These data indicate the existence of some level of immunological cross-reactivity between these two adenovirus subgroups. Based on these results, it is suggested that future studies should undertake similarly stringent analyses of alternative Ad serotypes to establish their effectiveness as replacements for Ad5.     

血清NGF、BDNF、GFAP水平与老年血管性痴呆严重程度的相关性分析

目的:探讨血清神经生长因子(Nerve Growth Factor,NGF)、脑源性神经营养因子(Brain Derived Neural Nutrition Factor, BDNF)、胶质纤维酸性蛋白(Glial Fibrillary Acidic Protein,GFAP)水平与老年血管性痴呆严重程度的相关性。方法:选择我院2016年1月至2018年12月收治的81例老年血管性痴呆患者,根据简易精神状态检查表(MMES)评分将其分为三组,以MMSE评分21～26分者为轻度组(26例),10～20分者为中度组(28例),0～9分者为重度组(27例),同时选择来院体检的50例健康者作为对照组,检测和比较各组的血清NGF、BDNF、GFAP水平,分析血清NGF、BDNF、GFAP水平与老年血管性痴呆患者MMSE分值的相关性。结果:老年血管性痴呆者的血清NGF、GFAP水平明显高于对照组(P0.05),血清BDNF水平明显低于对照组(P0.05)。轻度组、中度组、重度组的血清NGF、GFAP、BDNF水平对比差异均有统计学意义(P0.05):NGF水平:轻度组中度组重度组,BDNF水平:轻度组中度组重度组;GFAP水平:轻度组中度组重度组。血清NGF、BDNF水平与MMSE分值评分呈显著正相关(r_1=0.652,r_2=0.671,P0.05),血清GFAP水平与MMSE分值呈显著负相关(r3=-0.681,P0.05)。结论:血清NGF、BDNF、GFAP水平均与老年血管性痴呆的严重程度密切相关,可能用于评估老年血管性痴呆病情的严重程度。     

高危型HPV和L1壳蛋白与宫颈病变关系的研究进展

宫颈病变是女性最常见的疾病之一,是多种因素共同作用的结果。宫颈癌及癌前病变的发生、发展是一个多因素、多步骤的复杂过程,多种基因的改变引发细胞的增殖失控。大量资料表明高危型人乳头瘤病毒(HPV)感染是宫颈病变的主要危险因素,其中HPV16、18型感染占了绝大部分。HPV整合状态与宫颈病变程度密切相关,人乳头瘤病毒(HPV)L1壳蛋白为免疫杀伤HPV病毒的主要靶位,L1蛋白的表达有利于激发人体细胞免疫,清除感染的细胞。HPV感染机体且病毒处于复制阶段时L蛋白才在机体中表达,但是当HPV病毒DNA与宿主细胞基因整合后,L1壳蛋白将不表达,无法形成一系列免疫反应,引发宫颈癌。HPV L1壳蛋白的表达缺失与宫颈病变的进展密切相关。本文对高危型HPV与HPV L1壳蛋白在宫颈病变中的研究进展作一综述。     

Dominant Mutations in GRHL3 Cause Van der Woude Syndrome and Disrupt Oral Periderm Development

Mutations in interferon regulatory factor 6 (IRF6) account for ∼70% of cases of Van der Woude syndrome (VWS), the most common syndromic form of cleft lip and palate. In 8 of 45 VWS-affected families lacking a mutation in IRF6, we found coding mutations in grainyhead-like 3 (GRHL3). According to a zebrafish-based assay, the disease-associated GRHL3 mutations abrogated periderm development and were consistent with a dominant-negative effect, in contrast to haploinsufficiency seen in most VWS cases caused by IRF6 mutations. In mouse, all embryos lacking Grhl3 exhibited abnormal oral periderm and 17% developed a cleft palate. Analysis of the oral phenotype of double heterozygote (Irf6^+/−;Grhl3^+/−) murine embryos failed to detect epistasis between the two genes, suggesting that they function in separate but convergent pathways during palatogenesis. Taken together, our data demonstrated that mutations in two genes, IRF6 and GRHL3, can lead to nearly identical phenotypes of orofacial cleft. They supported the hypotheses that both genes are essential for the presence of a functional oral periderm and that failure of this process contributes to VWS.     

ARP2/3 complex‐mediated actin dynamics is required for hydrogen peroxide‐induced stomatal closure in Arabidopsis

Multiple cellular events like dynamic actin reorganization and hydrogen peroxide (H₂O₂) production were demonstrated to be involved in abscisic acid (ABA)‐induced stomatal closure. However, the relationship between them as well as the underlying mechanisms remains poorly understood. Here, we showed that H₂O₂ generation is indispensable for ABA induction of actin reorganization in guard cells of Arabidopsis that requires the presence of ARP2/3 complex. H₂O₂‐induced stomatal closure was delayed in the mutants of arpc4 and arpc5, and the rate of actin reorganization was slowed down in arpc4 and arpc5 in response to H₂O₂, suggesting that ARP2/3‐mediated actin nucleation is required for H₂O₂‐induced actin cytoskeleton remodelling. Furthermore, the expression of H₂O₂ biosynthetic related gene AtrbohD and the accumulation of H₂O₂ was delayed in response to ABA in arpc4 and arpc5, demonstrating that misregulated actin dynamics affects H₂O₂ production upon ABA treatment. These results support a possible causal relation between the production of H₂O₂ and actin dynamics in ABA‐mediated guard cell signalling: ABA triggers H₂O₂ generation that causes the reorganization of the actin cytoskeleton partially mediated by ARP2/3 complex, and ARP2/3 complex‐mediated actin dynamics may feedback regulate H₂O₂ production.     

应激刺激对大鼠下丘脑血管升压素mRNA水平的影响

实验在给予慢性应激刺激的Ｓｐｒａｇｕｅ Ｄａｗｌａｙ雄性大鼠中进行,用核酸斑点杂交技术观察大鼠下丘脑组织中血管升压素（ＡＶＰ）ｍＲＮＡ水平变化,用异羟基洋地黄毒甙（ＤＩＧ）标记的２６个碱基长寡聚核苷酸作为检测探针。结果观察到：在给予电击足底结合噪声的应激刺激之后,尾动脉收缩压逐渐升高,到刺激第５天,刺激组与对照组动物之间尾动脉收缩压差别有统计学显著意义;到刺激第９天,刺激组尾动脉收缩压达最高值;以     

低能离子束促进小麦copia-反转录转座子的转录与转座

LTR-反转录转座子是真核基因组重要的组成部分,能通过RNA中间体完成在基因组上的转座。小麦种子受低能氮离子束注入后能促进其反转录转座子的转录,经不同剂量的氮离子束注入的种子发芽24h和48h后,其中的copia-反转录转座子的转录都有不同程度的提高,最高可达到对照的40倍。用基于反转录转座子扩增多态性和反转录转座子微卫星扩增多态性2种分子标记技术扩增经低能氮离子注入后的小麦DNA,指纹图谱显示出了一定的多态性,这说明低能氮离子束激活了小麦中反转录转座子的转座活性。转座子转录活性的增强可调节其邻近基因的表达和mRNA的拼接,反转录转座子插入到基因组上新的位点后,使基因组上的基因发生了重排,这种重排可能表现为植物表型的改变。因此,推测反转录转座子的转录活性提高和转座激活是产生低能离子束注入生物效应的重要原因之一。     

KILLER WHALE (ORCINUS ORCA) AUDITORY EVOKED POTENTIALS TO RHYTHMIC CLICKS

Temporal auditory mechanisms were measured in killer whales ( Orcinus orca ) by recording auditory evoked potentials (AEPs) to clicks. Clicks were presented at rates from 10/sec to 1,600/sec. At low rates, clicks evoked an AEP similar to the auditory brainstem response (ABR) of other odontocetes; however, peak latencies of the main waves were 3–3.7 msec longer than in bottlenose dolphins. Fourier analysis of the ABR showed a prominent peak at 300–400 Hz and a smaller one at 800–1,200 Hz. High-rate click presentation (more than 100/sec) evoked a rate-following response (RFR). The RFR amplitude depended little on rate up to 400/sec, decreased at higher rates and became undetectable at 1,120/sec. Fourier analysis showed that RFR fundamental amplitude dependence on frequency closely resembled the ABR spectrum. The fundamental could follow clicks to around 1,000/sec, although higher harmonics of lower rates could arise at frequencies as high as 1,200 Hz. Both RFR fundamental phase dependence on frequency and the response lag after a click train indicated an RFR group delay of around 7.5 msec. This corresponds to the latency of ABR waves PIII-NIV, which indicates the RFR originates as a rhythmic, overlapping ABR sequence. The data suggest the killer whale auditory system can follow high click rates, an ability that may have been selected for as a function of high-frequency hearing and the use of rapid clicks in echolocation.     

水稻不同生育期茎秆生化成分的变化及其与抗倒伏能力的关系

以抗倒伏品种‘南粳44’、‘武运粳7号’与不抗倒伏品系‘宁7412’为试验材料,通过对水稻不同生育期茎秆硅、钾、钙、镁含量及可溶性糖含量的测定,结合氮钾肥配比试验,研究了水稻不同生育期茎秆硅、钾、钙、镁含量及可溶性糖含量的变化及其与茎秆抗倒伏能力的关系。结果表明：水稻茎秆的硅、钙、镁含量及可溶性糖含量随生育进程呈上升趋势,而茎秆的钾含量呈现先降后升的趋势。在不同施肥水平条件下,水稻茎秆的硅、钾、钙、镁含量及可溶性糖含量存在一定的差异。抗倒伏品种‘南粳44’和‘武运粳7号’茎秆基部抗折力较强,施肥对水稻茎秆基部抗折力有一定的影响。水稻生殖生育期茎秆的硅含量、可溶性糖含量与茎秆基部抗折力呈极显著正相关（P〈0.01）,与水稻抗倒伏能力的强弱有一定的关系。