Live attenuated malaria vaccine designed to protect through hepatic CD8⁺ T cell immunity

Our goal is to develop a vaccine that sustainably prevents Plasmodium falciparum (Pf) malaria in ≥80% of recipients. Pf sporozoites (PfSPZ) administered by mosquito bites are the only immunogens shown to induce such protection in humans. Such protection is thought to be mediated by CD8(+) T cells in the liver that secrete interferon-γ (IFN-γ). We report that purified irradiated PfSPZ administered to 80 volunteers by needle inoculation in the skin was safe, but suboptimally immunogenic and protective. Animal studies demonstrated that intravenous immunization was critical for inducing a high frequency of PfSPZ-specific CD8(+), IFN-γ-producing T cells in the liver (nonhuman primates, mice) and conferring protection (mice). Our results suggest that intravenous administration of this vaccine will lead to the prevention of infection with Pf malaria.     

不同苗龄毛棉杜鹃幼苗与杜鹃类菌根真菌的共生效应

[目的]分析接种杜鹃类真菌对不同苗龄毛棉杜鹃(Rhododendron moulmainense)的共生效应,为毛棉杜鹃菌根化育苗提供理论依据.[方法]设大苗龄(b,1.5年生)、中苗龄(m,1.0年生)和小苗龄(s,0.5年生)3种毛棉杜鹃实生苗苗龄,通过回接试验,测定分析接种福廷瓶头霉菌(Phialocephala fortinii,Pf)和聚多曲霉菌(Aspergillus sydowii,As)的毛棉杜鹃幼苗生长、根系形态和叶绿素荧光参数等指标.[结果]与不接种任何真菌对照(CK)相比,接种Pf和As两种菌种对大、中苗龄毛棉杜鹃幼苗生长均有促进作用,接种As对大、中苗龄地上或地下部的促进作用均小于接种Pf.其中,接种Pf对中苗龄毛棉杜鹃幼苗地上部生长的促进作用最明显,对大苗龄毛棉杜鹃幼苗地上部生长的促进作用次之;接种Pf对大苗龄毛棉杜鹃幼苗根系生长的促进作用最明显,对中苗龄毛棉杜鹃幼苗根系生长的促进作用次之;接种Pf和As对小苗龄毛棉杜鹃幼苗地上部及根系生长表现抑制作用.3个苗龄接菌组毛棉杜鹃幼苗的比叶面积均小于CK组;接菌组的实际光量子产量(YII)、非光化学淬灭(NPQ)和表观光合电子传递速率(ETR)均显著高于CK组(P<0.05),各苗龄相对于CK组提高倍数排序为小苗龄>中苗龄>大苗龄.总体上,接种Pf对毛棉杜鹃幼苗光合能力的提高作用优于接种As.[结论]接种Pf和As可与大、中和小苗龄毛棉杜鹃幼苗形成良好的共生体,其中,大苗龄和中苗龄接种Pf和As后可提高其光合能力,促进植株和根系生长,尤其以接种Pf的促进作用更明显,小苗龄接种Pf和As后生长受到一定的抑制.因此,毛棉杜鹃的菌根化育苗以选择中苗龄接种Pf效果更佳.     

Chemical synthesis of poliovirus cDNA: generation of infectious virus in the absence of natural template

Full-length poliovirus complementary DNA (cDNA) was synthesized by assembling oligonucleotides of plus and minus strand polarity. The synthetic poliovirus cDNA was transcribed by RNA polymerase into viral RNA, which translated and replicated in a cell-free extract, resulting in the de novo synthesis of infectious poliovirus. Experiments in tissue culture using neutralizing antibodies and CD155 receptor-specific antibodies and neurovirulence tests in CD155 transgenic mice confirmed that the synthetic virus had biochemical and pathogenic characteristics of poliovirus. Our results show that it is possible to synthesize an infectious agent by in vitro chemical-biochemical means solely by following instructions from a written sequence.     

The function of a leader peptide in translocating charged amino acyl residues across a membrane

Insertion of bacteriophage coat proteins into the membrane of infected bacterial cells can be studied as a model system of protein translocation across membranes. The coat protein of the filamentous bacteriophage Pf3--which infects Pseudomonas aeruginosa--is 44 amino acids in length and has the same basic structure as the coat protein of bacteriophage M13, which infects Escherichia coli. However, unlike the Pf3 coat protein, the M13 coat protein is synthesized as a precursor (procoat) with a typical leader (signal) sequence, which is cleaved after membrane insertion. Nevertheless, when the gene encoding the Pf3 coat protein is expressed in E. coli, the protein is translocated across the membrane. Hybrid M13 and Pf3 coat proteins were constructed in an attempt to understand how the Pf3 coat protein is translocated without a leader sequence. These studies demonstrated that the extracellular regions of the proteins determined their cellular location. When three charged residues in this region were neutralized, the leader-free M13 coat protein was also inserted into the membrane. Differences in the water shell surrounding these residues may account for efficient membrane insertion of the protein without a leader sequence.     

亲和性组合中拮抗细菌菌株Pf7-14和P13-R在稻株上的种群动态研究

拮抗细菌菌株(Pseudom onas fluorescens)Pf7-14,(Pseudom onas sp.)P13,均显著地抑制水稻纹枯病。离体测定表明,两菌株并不相互抑制,其为一个亲和性的组合。在温室条件下(菲律宾国际水稻研究所),通过分批播种,接种纹枯病菌(Rhizoctonia solani),在水稻分孽盛期喷雾拮抗细菌,并采用定期取样,平板稀释分离法在含不同抗生素的培养基中回收细菌,测定该组合Pf7-14 P13-R(Pf7-14是天然的抗萘啶酮酸菌株;P13-R是P13的利福平抗性突变体)中菌株Pf7-14和P13-R以及单剂菌株Pf7-14和P13-R在水稻健株及纹枯病株上的种群数量。2批试验结果均表明,混剂中菌株Pf7-14和P13-R的种群动态的变化是不一致的。在健株上,在相同的时间内单、混剂中菌株Pf7-14的种群数量基本保持相等;但在病株上,混剂中菌株Pf7-14的种群数量在应用后的一段时间内却显著地高于单剂的群体数量,第1批试验中,显著性差异发生在拮抗细菌应用后1~4 d,单、混剂中菌株Pf7-14群体数量相差可高达3.99倍,第2批试验中,显著性差异发生在应用后4~7 d,群体相差可高达3.17倍。相比,无论是在健株还是在病株上,在相同的时间内单、混剂中菌株P13的种群数量基本保持大致相等,没有显著性差异。这些结果表明混剂中菌株Pf7-14比P13在病斑上具有更强的营养竞争力。同时表明,理论上这个亲和性的拮抗细菌组合是一个增效的组合。     

Binding of HTLV-III/LAV to T4+ T cells by a complex of the 110K viral protein and the T4 molecule

Human T-lymphotropic virus type III (HTLV-III) or lymphadenopathy-associated virus (LAV) is tropic for human T cells with the helper-inducer phenotype, as defined by reactivity with monoclonal antibodies specific for the T4 molecule. Treatment of T4+ T cells with monoclonal antibodies to T4 antigen blocks HTLV-III/LAV binding, syncytia formation, and infectivity. Thus, it has been inferred that the T4 molecule itself is a virus receptor. In the present studies, the surfaces of T4+ T cells were labeled radioactively, and then the cells were exposed to virus. After the cells were lysed, HTLV-III/LAV antibodies were found to precipitate a surface protein with a molecular weight of 58,000 (58K). By blocking and absorption experiments, this 58K protein was identified as the T4 molecule. No cell-surface structures other than the T4 molecule were involved in the antibody-antigen complex formation. Two monoclonal antibodies, each reactive with a separate epitope of the T4 molecule, were tested for their binding capacities in the presence of HTLV-III/LAV. When HTLV-III/LAV was bound to T4+ T cells, the virus blocked the binding of one of the monoclonal antibodies, T4A (OKT4A), but not of the other, T4 (OKT4). When HTLV-III/LAV was internally radiolabeled and bound to T4+ T cells which were then lysed, a viral glycoprotein of 110K (gp110) coprecipitated with the T4 molecule. The binding of gp110 to the T4 molecule may thus be a major factor in HTLV-III/LAV tropism and may prove useful in developing therapeutic or preventive measures for the acquired immune deficiency syndrome.     

Short-term cyclic variations and diurnal variations of the venus upper atmosphere

Measurements of satellite drag obtained from the orbital decay of the Pioneer Venus orbiter on the nightside of Venus indicate an atomic oxygen atmosphere near 155 kilometers (an order of magnitude less dense than expected) with nighttime inferred exospheric temperatures averaging as low as 110 K. Densities at these altitudes decrease sharply from day to night, contrary to the predicted nighttime oxygen bulge. This decrease may be indicative of an unexpectedly weak transport across the evening terminator or a very strong heat sink at night that is possibly related to vertical eddy heat transport. Large periodic oscillations in density and inferred exospheric temperature are detected with a period of 5 to 6 days. We have subsequently discovered temperature variations of the same period in the stratosphere, which are tentatively interpreted as planetary-scale waves that may propagate upward producing the periodic variations in the thermosphere and exosphere. The peak-to-peak amplitude of the temperature oscillations associated with these waves apparently increases with altitude approximately as follows: 1 K (70 kilometers), 3 K (90 kilometers), 40 K (155 kilometers). Inferred nighttime exospheric temperatures are found to be asymmetric relative to midnight, minimizing on the morning side. The possibility of superrotation of the thermosphere, and exosphere is discussed.     

Reshaping human antibodies: grafting an antilysozyme activity

The production of therapeutic human monoclonal antibodies by hybridoma technology has proved difficult, and this has prompted the "humanizing" of mouse monoclonal antibodies by recombinant DNA techniques. It was shown previously that the binding site for a small hapten could be grafted from the heavy-chain variable domain of a mouse antibody to that of a human myeloma protein by transplanting the hypervariable loops. It is now shown that a large binding site for a protein antigen (lysozyme) can also be transplanted from mouse to human heavy chain. The success of such constructions may be facilitated by an induced-fit mechanism.     

黄颡鱼源柱状黄杆菌的分离鉴定及其对翘嘴鳜的致病性

从患病黄颡鱼上分离出1株柱状黄杆菌(Pf1),对其进行分类鉴定和药物敏感试验。用Pf1感染黄颡鱼和翘嘴鳜,发现Pf1对2种鱼均有致病性。Pf1可导致翘嘴鳜多个组织细胞坏死和炎症反应,严重损伤肝、体肾和鳃。被感染翘嘴鳜肝细胞肿胀、空泡化坏死;肾小管广泛坏死和大量炎症细胞浸润;鳃小片毛细血管充血,呼吸上皮细胞肿胀变性,鳃的呼吸功能衰退。鳃部病变导致感染鳜呼吸频率加快、游动缓慢,最后大量死亡。     

Cloned gene of Rickettsia rickettsii surface antigen: candidate vaccine for Rocky Mountain spotted fever

Two major protective antigens of Rickettsia rickettsii have been previously described. In this study, we cloned the gene encoding one of these antigens into Escherichia coli and tested the effectiveness of the recombinant-made product as a vaccine for Rocky Mountain spotted fever. A clone bank of R strain R. rickettsii DNA was made in E. coli K-12 by using the plasmid vector pBR322. Transformants were screened for their ability to make rickettsial antigens by reactivity with rabbit antibodies to R. rickettsii. One of the transformants, EM24(pGAM21), made a product reactive with two monoclonal antibodies that recognize a 155-kilodalton protein of R. rickettsii. One of the monoclonal antibodies was a member of a class of antibodies that react to heat-sensitive epitopes and protect mice injected with a potentially lethal dose of viable R. rickettsii. The cloned product contained this protective heat-sensitive epitope. In order to obtain enhanced expression, the gene was subcloned downstream of the lactose promoter on the plasmid vector pUC8. A sonic lysate of E. coli harboring the pUC8 subclone was used successfully as a vaccine to protect mice injected with a lethal dose of the viable R. rickettsii.     

基于SNP标记的玉米自交系S155、PHA458和A01的遗传多样性分析

为了分析玉米品种新科891的遗传结构和遗传多样性,本研究利用Affymetrix Axiom 10K SNP芯片对玉米品种新科891的双亲自交系（母本S155、父本PHA458）及自交系A01进行全基因组扫描,获得多态性标记4 926个。结果表明,群体聚类分析和群体遗传结构分析的杂种优势类群划分结果基本一致。母本S155为SS类种质,父本PHA458含有NSS和塘四平头类血缘,自交系A01为塘四平头类血缘,新科891的杂种优势模式为SS×NSS,这为新科891双亲的后续改良与创新奠定基础,并为杂交组配的育种工作提供依据。     

高效溶磷微生物菌株的筛选、鉴定及其对磷素效率的影响

从我省中部地区种植玉米的贫磷土壤样品进行分离筛选获得了综合能力较强的细菌24株;通过DNA测序和16S rRNA系统发育分析,分别鉴定为枯草芽孢杆菌(Bacillus subtilis strain Q1)、苏云金芽胞杆菌(Bacillus thuringiensis serovar kurstaki strain AR-10)、荧光假单胞菌(Pseudomonas fluorescens Pf0-1)、蜡状芽孢杆菌(Bacillus cereus)、伯克金氏菌(Burkholderia sp.CEB01056)、Oxalobacteraceae bacterium NR186和肠杆菌(Enterobacter ludwigii strain K9);田间试验结果表明:溶磷发酵液与化肥配施的情况下,磷肥用量减少1/3或2/3的处理与全量磷肥产量差异不显著,各个处理与对照相比均表现明显的增产效应。     

The human immune response to the OKT3 monoclonal antibody is oligoclonal

The availability of highly specific and homogeneous antibodies to human T cells by the hybridoma technique has elicited new interest in the clinical use of antibodies to lymphocytes as immunosuppressive agents. OKT3 is the murine monoclonal antibody that has been the most widely used in clinical transplantation to induce immunosuppression. This antibody recognizes a membrane molecular complex, exclusively present on mature human T lymphocytes, which is tightly linked to the T-cell antigen receptor. The long-term therapeutic use of murine monoclonal antibodies in vivo is hampered by the intense antibody response that occurs in most human patients. Thus, when administered alone, OKT3 manifests its immunosuppressive activity only during the 10 to 15 days that precede the onset of sensitization. The results presented here show, by use of isoelectrofocusing, that the antibody response to OKT3, already reported to be restricted in its specificity (only anti-isotypic and anti-idiotypic antibodies are produced), is in addition oligoclonal. This restriction of the anti-monoclonal response may suggest that an efficient way to circumvent the sensitization problem would be to administer consecutively different monoclonal antibodies presenting the same specificity but distinct idiotypes.     

Broadly neutralizing antibodies present new prospects to counter highly antigenically diverse viruses

Certain human pathogens avoid elimination by our immune system by rapidly mutating the surface protein sites targeted by antibody responses, and consequently they tend to be problematic for vaccine development. The behavior described is prominent for a subset of viruses--the highly antigenically diverse viruses--which include HIV, influenza, and hepatitis C viruses. However, these viruses do harbor highly conserved exposed sites, usually associated with function, which can be targeted by broadly neutralizing antibodies. Until recently, not many such antibodies were known, but advances in the field have enabled increasing numbers to be identified. Molecular characterizations of the antibodies and, most importantly, of the sites of vulnerability that they recognize give hope for the discovery of new vaccines and drugs.     

Naturally occurring antibodies reactive with sperm proteins: apparent deficiency in AIDS sera

A set of naturally occurring immunoglobulin M (IgM) antibodies that are reactive with a defined subset of proteins in the acrosomal cap region of human sperm has been identified. These antibodies are present in a broad spectrum of human sera from males and females, 1 day to 40 years of age, and are absent or markedly deficient in a large proportion of sera from individuals with the acquired immune deficiency syndrome (AIDS) or at risk for AIDS. The subset of proteins with which the IgM antibodies are reactive includes a factor (or factors) capable of inhibiting lectin-induced T-lymphocyte proliferation. The prevalence of the sperm-reactive IgM antibodies indicates that they are not elicited by sperm. Further, immunoreactivity of the sperm proteins resulting in depletion of specific circulating IgM antibodies, or other interactions between the sperm proteins and elements of the immune system, may be a factor in the suppressed state of the immune system in AIDS.     

拮抗细菌菌株B5423-R和Pf7-14在水稻叶片上的定殖及其对土著细菌群体的影响

拮抗细菌菌株Bacillus subtilisB5423和Pseudomonas fluorescensPf 7-14能显著地抑制水稻纹枯病。在温室条件下,通过播种、叶片接种纹枯病菌Rhizoctonia solani,以及在水稻分蘖盛期喷雾菌株B5423-R(B5423的利福平抗性突变体)和Pf 7-14(天然的抗萘啶酮酸菌株)的细菌悬浮液,并通过定期取样、平板系列稀释法回收,监测了菌株B5423-R、Pf 7-14和土著细菌群体在水稻健叶和纹枯病叶上的种群动态。结果表明:在病斑叶片片段(病斑面积占70%~90%),在应用1~7 d后,菌株Pf 7-14的平均群体数量比在相同面积的健康叶片片段上显著性地高3~10倍,而菌株B5423-R的数量比在健康叶片片段上显著性地高12~50倍;土著细菌群体数量在病斑叶片上是健康叶片上的50~60倍。这些结果表明纹枯病病斑大大促进了土著细菌群体和引入的生防细菌的生长,且土著细菌群体和引入的生防细菌在营养和空间上是相互竞争的。     

Autoantibodies to a 64-kilodalton islet cell protein precede the onset of spontaneous diabetes in the BB rat

Spontaneous insulin-dependent diabetes mellitus (IDDM) in the BB rat is associated with the presence of antibodies to a 64-kilodalton rat islet cell protein. These protein antibodies appeared in young animals and remained for as long as 8 weeks before the clinical onset of IDDM. Antibodies to a 64-kilodalton human islet cell protein were found to be associated with human IDDM. Detection of the antibodies may therefore be used to predict an early immune reaction against pancreatic B cells.     

优质高产杂交中籼新组合辐优155的选育与应用研究

[目的]为杂交中籼新组合辐优155的推广应用提供参考.[方法]通过辐优155经安徽省2007、2008年2年区试和2009年生试,比较辐优155与对照品种Ⅱ优838主要农艺性状、产量、稻米品质及抗性,并分析了其大田栽培技术及制种技术要点,以探讨其生产潜力及推广价值.[结果]辐优155是安徽省铜陵县农业科科学研究所用辐88A与恢155配组育成的杂交中籼新组合,具有株型适中、穗大粒多、米质优、抗稻瘟病、中抗白叶枯病等特点,2010年通过安徽省农作物品种审定委员会审定.[结论]辐优155产量高,品质好,适宜在安徽全省种植.     

粉拟青霉南北株系、白僵菌的生理特性和毒力

粉拟青霉南北株系（Ｓ－Ｐｆ和Ｎ－Ｐｆ）、白僵菌（Ｂｂ）在２５℃下于水滴中的孢子萌发率分别为７９．８％、６７．２％和７８．４％，而当相对湿度为９０％以下时，孢子萌发率急剧下降．白僵菌孢子萌发率最适温度为２５℃，粉拟青霉均为２０℃；１５℃以上，Ｓ－Ｐｆ孢子萌发率高于Ｎ－Ｐｆ，而１５℃以下时则相反．在ＰＤＡ培养基上，粉拟青霉南北株系产孢量基本相同；Ｎ－Ｐｆ最适生长温度、产孢温度为２０～２５℃；Ｓ－Ｐｆ则为２０℃．在２０℃以上，白僵菌对马尾松尾虫感染率高于粉拟青霉；而在１５℃以下，粉拟青霉则明显高于白僵菌．     

抗猪大肠杆菌功能性卵黄粉的制备及对小鼠的攻毒保护作用

通过制备抗致病性大肠杆菌特异性IgY及富含特异性IgY的功能性卵黄粉,进行小鼠的攻毒保护试验,探讨功能性卵黄粉对由致病性大肠杆菌引起的疾病的防治作用.结果显示:(1)致病性大肠杆菌与特异性IgY混合后,菌体出现凝集,且病原菌的运动力明显受抑制;(2)浓度为1.3×109CFU·mL-1的致病性大肠杆菌K88、K99和9...