含变形链球菌PAcA基因的真核表达质粒pcDNA3.1-PAcA的构建与鉴定

目的 :构建含有免疫刺激序列、可防止变形链球菌在牙面粘附的DNA疫苗。方法 :利用PCR技术由质粒pPAcA CTA2 B扩增编码PAcA的DNA片段 ;通过T A克隆技术将目的DNA片段克隆于载体pMD1 8 T ,鉴定插入方向后 ,将目的DNA从载体上释放 ,再克隆于含有CpG免疫刺激序列的真核表达载体pcDNA3 .1 ,构建出用作防龋疫苗的真核表达质粒pcDNA3 .1 PAcA。结果 :通过对重组质粒pcDNA3 .1 PAcA进行酶切图谱和DNA序列测定分析 ,证明真核表达重组质粒pcDNA3 .1 PAcA构建成功、阅读框架正确。结论 :利用T A克隆等技术可成功将编码PAcA的DNA片段克隆到含有免疫刺激序列CpG的真核表达载体pcDNA3 .1 ,构建出真核表达重组质粒pcDNA3 .1 PAcA。     

HIV-1 gp41 heptad repeat 2 (HR2) possesses an amino acid domain that resembles the allergen domain in <Emphasis Type="Italic">Aspergillus fumigatus Asp f1</Emphasis> protein: review,hypothesis and implications

Enfuvirtide (ENF, T-20, Fuzeon) is the first synthetic peptide to be modeled according to the amino acid sequence of HIV-1 heptad repeat 2, which was used to treat cohorts of HIV-1-infected individuals who had failed to respond to treatment with the anti-HIV-1 cocktail HAART. It was reported that when injected subcutaneously, Enfuvirtide reduced viral RNA in patients’ blood by 1.96 log₁₀, leading to a subsequent increase in the number of CD4⁺ T cells in the blood. The drug treatment caused adverse effects at the injection site in a small number of treated individuals, and a gradual increase in IgE in the blood during prolonged treatment. Enfuvirtide was approved for treatment of HIV-1 patients who developed resistance to HAART. The present review attempts to explain the adverse effects of Enfuvirtide at the skin site of injection, and the gradual increase in IgE in patients’ blood during treatment. These phenomena were reported to resemble the effect of allergens that cause asthma in humans. It is hypothesized that since the amino acid domain of the Asp f1 allergen from Aspergillus fumigatus was identified in the N-terminus of an 18 kDa protein, it may be useful to compare Asp f1 peptide aa 7–22 from the β-hairpin sequence to the β-hairpin sequence of the heptad repeat 2 of HIV-1 gp41. The comparison revealed that the amino acid sequence resembles part of the Asp f1 aa 7–22 allergenic domain. The heptad repeat 1 of gp41 also resembles the fungal allergen. It is suggested that the Enfuvirtide peptide be tested experimentally to determine if ENF peptide is capable of binding to IgE antibodies from Enfuvirtide-treated, HIV-1-infected patients, and whether the HR2-derived peptide is capable of inducing basophils that were isolated from healthy individuals and from ENF-treated and untreated HIV-1 patients to release histamine and IL-4.     

Mutational analysis of the GDD sequence motif of classical swine fever virus RNA-dependent RNA polymerases

To define the function of the GDD motif of the RNA-dependent RNA polymerase (RdRp) of classical swine fever virus (CSFV), single amino acid substitutions were introduced into the CSFV NS5B. All substitutions within the GDD motif were detrimental to the polymerase activity, the binding activity and the terminal nucleotidyl transferase activity of the NS5B protein. It was also found that the wild-type NS5B had higher RdRp activity with Mg(+2) than with Mn(+2) whereas some mutants worked better with Mn(+2) than with Mg(+2), suggesting that substitutions within the GDD motif modified the enzyme cation preferences and the GDD sequence of CSFV NS5B might be involved in polymerase-metal interaction. Therefore, the GDD amino acid sequence is important for the function of CSFV RdRp.     

Natural killer cell cytotoxicity and its regulation by inhibitory receptors

Natural killer (NK) cells express an array of germ‐line encoded receptors that are capable of triggering cytotoxicity. NK cells tend to express many members of a given family of signalling molecules. The presence of many activating receptors and many members of a given family of signalling molecules can enable NK cells to detect different kinds of target cells, and to mount different kinds of responses. This contributes also to the robustness of NK cells responses; cytotoxic functions of NK cells often remain unaffected in the absence of selected signalling molecules. NK cells express many MHC‐I‐specific inhibitory receptors. Signals from MHC‐I‐specific inhibitory receptors tightly control NK cell cytotoxicity and, paradoxically, maintain NK cells in a state of proper responsiveness. This review provides a brief overview of the events that underlie NK cell activation, and how signals from inhibitory receptors intercept NK cell activation to prevent inappropriate triggering of cytotoxicity.     

Mechanisms and biomarkers of immune quiescence in kidney transplantation

This review discusses the current understanding of biomarkers of immune quiescence based on reviews of published literature in kidney transplant operational tolerance and mechanistic studies based on a better characterization of the stable, well-functioning renal allograft.     

血清趋化因子配体5预测膀胱癌外科治疗后复发的价值

目的探讨血清趋化因子配体5(CXCL5)对膀胱癌外科治疗后复发的预测价值。方法选取行膀胱癌根治术的98例患者,根据病理分期分为肌层浸润性膀胱癌(MIBC)组和非肌层浸润性膀胱癌(NMIBC)组,比较2组的血清CXCL5水平和复发率,并分析所有患者术前血清CXCL5水平与其临床病理特征的关系。采用受试者工作曲线下面积(AUC)来评价其对膀胱癌复发的预测价值。结果 MIBC组的术前血清CXCL5水平均显著高于NMIBC组(P0.05)。年龄60岁、肿瘤位于膀胱底或膀胱体、M_1期、G_2~G_3级者的术前血清CXCL5水平显著升高(P0.05)。所有患者共随访6~36个月,复发36例(36.7%),MIBC组的复发率显著高于NMIBC组,复发者术前血清CXCL5水平显著高于无复发者(P0.05)。ROC曲线分析结果表明,术前血清CXCL5水平对MIBC、NMIBC的AUC为0.861(0.669~0.960,P0.05)、0.819(0.630~0.933,P0.05),其最佳临界值分别为828.9、589.3 pg/m L。结论术前血清CXCL5水平对膀胱癌患者术后复发有较好的预测价值。     

Acute inhibition of a cortical motor area impairs vocal control in singing zebra finches

Genetically targeted approaches that permit acute and reversible manipulation of neuronal circuit activity have enabled an unprecedented understanding of how discrete neuronal circuits control animal behavior. Zebra finch singing behavior has emerged as an excellent model for studying neuronal circuit mechanisms underlying the generation and learning of behavioral motor sequences. We employed a newly developed, reversible, neuronal silencing system in zebra finches to test the hypothesis that ensembles of neurons in the robust nucleus of the arcopallium (RA) control the acoustic structure of specific song parts, but not the timing nor the order of song elements. Subunits of an ivermectin‐gated chloride channel were expressed in a subset of RA neurons, and ligand administration consistently suppressed neuronal excitability. Suppression of activity in a group of RA neurons caused the birds to sing songs with degraded elements, although the order of song elements was unaffected. Furthermore some syllables disappeared in the middle or at the end of song motifs. Thus, our data suggest that generation of specific song parts is controlled by a subset of RA neurons, whereas elements order coordination and timing of whole songs are controlled by a higher premotor area.