Characteristics of Chimeric West Nile Virus Based on the Japanese Encephalitis Virus SA14-14-2 Backbone

West Nile virus disease (WND) is an arthropod-borne zoonosis responsible for nonspecific fever or severe encephalitis. The pathogen is West Nile virus belonging to the genus Flavivirus, family Flaviviridae. Every year, thousands of cases were reported, which poses significant public health risk. Here, we constructed a West Nile virus chimera, ChiVax-WN01, by replacing the prMΔE gene of JEV SA14-14-2 with that of the West Nile virus NY99. The ChiVax-WN01 chimera showed clear, different characters compared with that of JEV SA14-14-2 and WNV NY99 strain. An animal study indicated that the ChiVax-WN01 chimera presented moderate safety and immunogenicity for 4-week female BALB/c mice.     

Genetic and Phenotypic Properties of Vero Cell-Adapted Japanese Encephalitis Virus SA14-14-2 Vaccine Strain Variants and a Recombinant Clone,which Demonstrates Attenuation and Immunogenicity in Mice

The live-attenuated Japanese encephalitis virus (JEV) SA14-14-2 vaccine, produced in primary hamster kidney cells, is safe and effective. Past attempts to adapt this virus to replicate in cells that are more favorable for vaccine production resulted in mutations that significantly reduced immunogenicity. In this study, 10 genetically distinct Vero cell-adapted JEV SA14-14-2 variants were isolated and a recombinant wild-type JEV clone, modified to contain the JEV SA14-14-2 polyprotein amino acid sequence, was recovered in Vero cells. A single capsid protein mutation (S66L) was important for Vero cell-adaptation. Mutations were also identified that modulated virus sensitivity to type I interferon-stimulation in Vero cells. A subset of JEV SA14-14-2 variants and the recombinant clone were evaluated in vivo and exhibited levels of attenuation that varied significantly in suckling mice, but were avirulent and highly immunogenic in weanling mice and are promising candidates for the development of a second-generation, recombinant vaccine.     

Safety of a live-attenuated Japanese encephalitis virus vaccine (SA14-14-2) for children

In a study begun in 1985, 1,026 children between the ages of 5 and 12 years, living in an area of low Japanese encephalitis (JE) infection, were vaccinated with live-attenuated JE vaccine, strain SA14-14-2. A group of 47 of the vaccinated children, 5-6 years of age, were examined for fever and other systemic reactions every other day for 2 weeks following vaccination. None of these children had temperatures greater than 37.4 degrees C or other systemic reactions during the observation period. No untoward reactions were reported in the remainder of the vaccinated group. After immunization, seroconversion rates in seronegative children were 100% (GMT 35.3, n = 11), 100% (GMT 31.7, n = 12), and 83.3% (GMT 23, n = 10) in groups receiving vaccine diluted 1:3, 1:5, and 1:50, respectively. These results indicate that the JE SA14-14-2 live-attenuated vaccine is immunogenic and safe for children.     

流行性乙型脑炎病毒SA14-14-2株E蛋白基因在原核细胞中的高效表达及其表达产物的抗原性分析

目的　获得JEVE蛋白基因并使其在E coli中高效表达 ,以研究其抗原活性 ,为进一步研制ELISA早期诊断试剂盒奠定基础。方法　根据已发表的JEVSA - 14 - 14 - 2减毒株序列 ,设计一对特异性引物 ,通过RT -PCR扩增出E蛋白基因的cDNA片段 ,并将其克隆入 pET - 32a(+)表达载体中 ,构建重组融合表达载体pET -E ,转化大肠杆菌BL2 1(DE3)后 ,利用IPTG诱导获得高效表达。结果　扩增的E蛋白基因片段长 1113bp ,编码 371个氨基酸残基 ,该基因片段与国内发表的减毒株SA14 - 14 - 2碱基序列同源性为 10 0 %。表达产物分子量约为 6 2kD ,经Westernblotting分析表明表达产物具有较好的抗原性。结论　通过序列分析表明 ,我国的JEVSA - 14 - 14 - 2减毒株未发生基因突变。表达产物的稳定高效表达及其抗原特异性分析为今后ELISA早期诊断试剂盒的研制提供了依据。     

乙型脑炎病毒SA14-14-2株E基因的克隆、测序与表达

目的　克隆乙型脑炎病毒E基因并在大肠杆菌中进行表达 ,为今后研制乙脑分子诊断试剂盒和基因工程疫苗奠定基础。方法　根据乙型脑炎病毒株SA14 - 14 - 2E基因的序列设计一对引物 ,采用RT -PCR技术扩增其E基因全长cD NA ,将扩增产物克隆到pBlusecriptIISK(+)载体中 ,然后亚克隆到原核表达载体PGEX -KG中 ,筛选重组质粒 ,转化大肠杆菌BL2 1(DE3)表达重组蛋白。结果与结论　获得了含全长乙型脑炎病毒E基因的重组质粒 ,经酶切和测序证实构建正确。表达的目的蛋白经SDS -PAGE和Westernblot分析证实确为乙型脑炎病毒E蛋白且有生物学活性。     

三带喙库蚊胸腔接种乙型脑炎减毒活疫苗病毒的感染动态及致病力观察

目的通过蚊虫胸腔接种乙脑病毒减毒活疫苗SA14-14-2株,了解该疫苗病毒在蚊虫体内的繁殖情况及其毒力稳定性,进一步评价该疫苗的安全性。方法建立三带喙库蚊的实验室种群,用SA14-14-2、SA14和中山株胸腔接种三带喙库蚊,感染后不同时间取一定数量的蚊虫,研磨制成悬液,用空斑试验检测蚊虫体内的病毒滴度。用感染蚊悬液接种乳鼠和感染蚊虫直接叮咬乳鼠的方法,观察对乳鼠的致病性。结果SA14-14-2、SA14和中山株病毒感染蚊虫后,第2~20d蚊虫体内均能检测到病毒,其中SA14-14-2株的滴度为2~3.72 logPFU/ml,SA14株为3~4.85 logPFU/ml,中山株为3~5.40 log-PFU/ml。中山株的感染滴度最高,其次是SA14株,SA14-14-2株的感染滴度最低,表明蚊虫对野毒株(中山株和SA14株)更为敏感。感染SA14-14-2病毒的三带喙库蚊悬液接种乳鼠,未能引起乳鼠发病或死亡。感染SA14-14-2病毒的三带喙库蚊叮咬乳鼠,未见乳鼠发病或死亡,也未从小白鼠血清中检测到乙脑病毒抗体。结论经胸腔接种,SA14-14-2病毒能在三带喙库蚊体内稳定繁殖。动物接种和蚊虫叮咬试验表明,经蚊体内繁殖的SA14-14-2病毒毒力仍保持原有的弱毒特性,表明该减毒活疫苗通过蚊虫体内繁殖后不会造成传播。     

Growth characteristics of the chimeric Japanese encephalitis virus vaccine candidate, ChimeriVax-JE (YF/JE SA14--14--2), in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes

The Japanese encephalitis (JE) virus vaccine candidate, ChimeriVax-JE, which consists of a yellow fever (YF) 17D virus backbone containing the prM and E genes from the JE vaccine strain JE SA14--14--2, exhibits restricted replication in non-human primates, producing only a low-level viremia following peripheral inoculation. Although this reduces the likelihood that hematophagous insects could become infected by feeding on a vaccinated host, it is prudent to investigate the replication kinetics of the vaccine virus in mosquito species that are known to vector the viruses from which the chimera is derived. In this study ChimeriVax-JE virus was compared to its parent viruses, as well as to wild-type JE virus, for its ability to replicate in Culex tritaeniorhynchus, Aedes albopictus, and Aedes aegypti mosquitoes. Individual mosquitoes were exposed to the viruses by oral ingestion of a virus-laden blood meal or by intrathoracic (IT) virus inoculation. ChimeriVax-JE virus did not replicate following ingestion by any of the three mosquito species. Additionally, replication was not detected after IT inoculation of ChimeriVax-JE in the primary JE virus vector, Cx. tritaeniorhynchus. ChimeriVax-JE exhibited moderate growth following IT inoculation into Ae. aegypti and Ae. albopictus, reaching titers of 3.6-5.0 log(10) PFU/mosquito. There was no change in the virus genotype associated with replication in mosquitoes. Similar results were observed in mosquitoes of all three species that were IT inoculated or had orally ingested the YF 17D vaccine virus. In contrast, all mosquitoes either IT inoculated with or orally fed wild-type and vaccine JE viruses became infected, reaching maximum titers of 5.4-7.3 log(10) PFU/mosquito. These results indicate that ChimeriVax-JE virus is restricted in its ability to infect and replicate in these mosquito vectors. The low viremia caused by ChimeriVax-JE in primates and poor infectivity for mosquitoes are safeguards against secondary spread of the vaccine virus.     

Attenuated metabolic effect of waist measurement in Japanese female patients with type 2 diabetes mellitus

Waist circumference (WC) was measured in 200 Japanese patients with type 2 diabetes mellitus (T2DM: male 106, female 94, mean age 61 years old) who had been admitted in our hospital, and relationship with various risk factors to predict future cardiovascular disease (CVD) was analyzed. There was a positive and statistically significant trend in WC levels with an increasing number of CVD risk factors in male patients, whereas no significant trend of WC was observed in female patients. The receiver operator characteristic (ROC) curve for WC to predict the presence of two or more risk factors of CVD depicted greater area under the curve in male patients (0.732) than that in female patients (0.571). Apart from positive correlation with fasting serum C-peptide (S-CPR) and log-transformed high-sensitivity C-reactive protein (log HS-CRP) in both genders, WC was positively correlated with log-transformed triglyceride (log TG), systolic and diastolic blood pressure (SBP and DBP) and negatively with HDL-cholesterol (HDL-C) in male patients, whereas it was negatively correlated with HbA1c and fasting plasma glucose (FPG) in female patients. The change of WC after administration (DeltaWC) was correlated with DeltaS-CPR, DeltaLDL-C, DeltaSBP and DeltaDBP in male patients, while no relationship was observed in female patients. In conclusion, WC is a reliable marker to predict future CVD events at least in Japanese male, but not female patients with T2DM.     

Intranasal Immunization with a Recombinant Avian Paramyxovirus Serotypes 2 Vector-Based Vaccine Induces Protection against H9N2 Avian Influenza in Chicken

Commercial inactivated vaccines against H9N2 avian influenza (AI) have been developed in China since 1990s and show excellent immunogenicity with strong HI antibodies. However, currently approved vaccines cannot meet the clinical demand for a live-vectored vaccine. Newcastle disease virus (NDV) vectored vaccines have shown effective protection in chickens against H9N2 virus. However, preexisting NDV antibodies may affect protective efficacy of the vaccine in the field. Here, we explored avian paramyxovirus serotype 2 (APMV-2) as a vector for developing an H9N2 vaccine via intranasal delivery. APMV-2 belongs to the same genus as NDV, distantly related to NDV in the phylogenetic tree, based on the sequences of Fusion (F) and hemagglutinin-neuraminidase (HN) gene, and has low cross-reactivity with anti-NDV antisera. We incorporated hemagglutinin (HA) of H9N2 into the junction of P and M gene in the APMV-2 genome by being flanked with the gene start, gene end, and UTR of each gene of APMV-2-T4 to generate seven recombinant APMV-2 viruses rAPMV-2/HAs, rAPMV-2-NPUTR-HA, rAPMV-2-PUTR-HA, rAPMV-2-FUTR-HA, rAPMV-2-HNUTR-HA, rAPMV-2-LUTR-HA, and rAPMV-2-MUTR-HA, expressing HA. The rAPMV-2/HAs displayed similar pathogenicity compared with the parental APMV-2-T4 virus and expressed HA protein in infected CEF cells. The NP-UTR facilitated the expression and secretion of HA protein in cells infected with rAPMV-2-NPUTR-HA. Animal studies demonstrated that immunization with rAPMV-2-NPUTR-HA elicited effective H9N2-specific antibody (6.14 ± 1.2 log2) responses and conferred complete immune protection to prevent viral shedding in the oropharyngeal and cloacal swabs from chickens challenged with H9N2 virus. This study suggests that our recombinant APMV-2 virus is safe and immunogenic and can be a useful tool in the combat of H9N2 outbreaks in chicken.     

Deletion Mutants of the Attenuated Recombinant ASF Virus,BA71ΔCD2, Show Decreased Vaccine Efficacy

African swine fever (ASF) has become the major threat to the global swine industry. Lack of available commercial vaccines complicates the implementation of global control strategies. So far, only live attenuated ASF viruses (ASFV) have demonstrated solid protection efficacy at the experimental level. The implementation of molecular techniques has allowed the generation of a collection of deletion mutants lacking ASFV-specific virulence factors, some of them with promising potential as vaccine candidates against the pandemic genotype II ASFV strain currently circulating in Africa, Europe, Asia and Oceania. Despite promising results, there is room for improvement, mainly from the biosafety point of view. Aiming to improve the safety of BA71∆CD2, a cross-protective recombinant live attenuated virus (LAV) lacking the ASFV CD2v gene (encoding β-glucuronidase as a reporter gene) available in our laboratory, three new recombinants were generated using BA71∆CD2 as a template: the single mutant BA71∆CD2f, this time containing the fluorescent mCherry reporter gene instead of CD2v, and two double recombinants lacking CD2v and either the lectin gene (EP153R) or the uridine kinase (UK) gene (DP96R). Comparative in vivo experiments using BA71∆CD2f, BA71∆CD2DP96R and BA71∆CD2EP153R recombinant viruses as immunogens, demonstrated that deletion of either DP96R or EP153R from BA71∆CD2f decreases vaccine efficacy and does not improve safety. Our results additionally confirm ASFV challenge as the only available method today to evaluate the protective efficacy of any experimental vaccine. We believe that understanding the fine equilibrium between attenuation and inducing protection in vivo deserves further study and might contribute to more rational vaccine designs in the future.     

云南省乙型脑炎流行与防治现状调查

目的了解云南省乙型脑炎流行变化和趋势，为制定防治方案和策略提供科学依据。方法收集2002～2005年云南省乙脑疫情资料，采用乙型脑炎病例个案凋查表进行现场既往流行病学个案调查，应用诱蚊灯和吸蚊管人工捕蚊方法进行现场传播媒介调查，同时进行防治现状调查。结果2002～2004年全省报告乙脑病例1272例，发病率在1‰～1．84‰的县市有8个，每年平均有69个县（市）有病例报告，多集中在6～9月发病。2005年全省报告乙脑病例426例，比2004年397例上升12．40％；死亡22例，上升69．23％，病死率5．16％，上升50．44％。全省16个州（市）中有15个州（市）有乙脑病例报告。既往流行病学个案调查6个县（市）17例，其中儿童发病12例（占70．59％），治疗后有严重后遗症4例（致残率23．53％），死亡4例（病死率23．53％）；媒介调查12个县（市），乙脑主要传播媒介三带喙库蚊为当地的主要优势蚊种（58．10％）；防治现状调查了15个州（市）26个县，主要采取的防治措施为：疫情监测、健康教育、疫点灭蚊喷洒、乙脑疫苗接种。其中疫苗预防接种调查8个州（市）26个县，年接种率在0．10％～9．58％之间。结论1）云南省乙型脑炎发病多、分布较广，儿童发病、致残、病死率较高；2）乙脑的主要传播媒介三带喙库蚊在云南省广泛分布；3）人群乙脑疫苗预防接种率较低，特别是农村儿童接种覆盖率非常低。     

Pharmacokinetics, prophylactic effects, and safety of a new recombinant FVIII formulated with sucrose (BAY 14-2222) in Japanese patients with hemophilia A

A clinical trial of BAY 14-2222, a recombinant factor VIII preparation (rFVIII) manufactured by new purification and formulation processes using sucrose as a stabilizer instead of human serum albumin, was performed in 5 previously treated Japanese patients with severe hemophilia A. In stage I, a single dose of BAY 14-2222 and Kogenate (a currently licensed rFVIII preparation) was administered alternately in the same patients to compare the pharmacokinetics of the 2 compounds using FVIII:C (FVIII clotting activity) as the measure of plasma drug levels. The normalized area under the curve (AUCnorm) and normalized maximal concentration (Cmax,norm) were slightly lower following the administration of BAY 14-2222 than those after the administration of Kogenate (ratio of BAY 14-2222/Kogenate:AUCnorm = 0.88, P = .050; and Cmax,norm = 0.87, P = .041). However, the biological half-life (t1/2) did not differ significantly between the 2 preparations (13.96 +/- 4.18 vs. 13.48 +/- 2.40 hours). The in vivo recovery of FVIII was 67.9 +/- 11.3% after the administration of BAY 14-2222 and 74.4 +/- 5.3% after the administration of Kogenate. In stage II, BAY 14-2222 was administered regularly to the 5 patients with hemophilia at single doses of 20 to 40 IU/kg 3 times weekly for 4 weeks, and its prophylactic effect on bleeding was evaluated. Results indicated that BAY 14-2222 has a good preventive effect on bleeding. Sixty-six infusions were performed in stages I and II of this trial, and no adverse reactions related to BAY 14-2222 were observed. In addition, there were no FVIII inhibitors or antibodies to foreign proteins detected. The trial confirmed that BAY 14-2222 is similar to Kogenate with respect to t1/2 and the in vivo recovery of FVIII:C and that periodic infusions for 4 weeks can be well tolerated. In addition, it was shown that BAY 14-2222 is effective in preventing bleeding. Thus it is expected that BAY 14-2222 will exhibit a hemostatic effect comparable to that of Kogenate in patients with hemophilia A.     

The metabolism of [1-14C]-, [2-14C]-, [3,4-14C]-, and [6-14C]- glucose in normal and diabetic polymorphonuclear leukocytes and during phagocytosis

Summary 1. Polymorphonuclear leucocytes from normal and diabetic subjects and from normal and alloxan diabetic rats were incubated with ¹⁴C-glucose, and allowed to phagocytize. — 2. The major ¹⁴CO₂ production originated from the pentose cycle. Approx. one third of ¹⁴CO₂ was formed by decarboxylation of pyruvate, whereas Krebs cycle activity was minimal. — 3. The pentose cycle metabolized 0.1–0.5% of the phosphorylated glucose, and phagocytosis increased this fraction 2–3 fold. No difference was found between normal and diabetic cells. — 4. The major endproduct of glucose metabolism was lactic acid. The randomization of ¹⁴C of glucose was not consistent with the sole operation of the oxidative or non-oxidative parts of the pentose cycle, but indicated the participation of a symmetrical C₃ intermediate, i.e. dihydroxy-acetone, in the metabolism of part of the glucose carbons. — 5. Very small amounts of ¹⁴C from glucose were found in aminoacids, metabolic acids not extractable with ether, and lipids. Appreciable amounts of ¹⁴C were, however, found in a compound (A), part of which precipitated as glyceroltribenzoate, and a compound (B), which presumably was of protein nature. The incorporation of ¹⁴C in compound A from normal leucocytes decreased during phagocytosis, and was insignificant in human diabetic cells. The incorporation of ¹⁴C from [1-¹⁴C]- and [2-¹⁴C]-, but not from [6-¹⁴C]-glucose, into compound B was greatly stimulated in normal cells during phagocytosis, but not so in diabetic leucocytes.

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Der Stoffwechsel von [1-¹⁴C]-, [2-¹⁴C]-, [3-¹⁴C]- und [6-¹⁴C]-Glucose in normalen und diabetischen polymorphkernigen Leukozyten und während der Phagozytose

Zusammenfassung 1. Polymorphkernige Leukozyten von Normalpersonen und Diabetikern und von normalen und alloxandiabetischen Ratten wurden mit ¹⁴C-Glucose inkubiert und zur Phagozytose angeregt. — 2. Der überwiegende Teil der ¹⁴CO₂ Produktion stammte aus dem Pentosezyklus. Etwa ein Drittel des ¹⁴CO₂ wurde bei der Decarboxylierung von Pyruvat gebildet, während die Aktivität des Krebs-Zyklus nur sehr gering war. — 3. Den Pentose-Phosphat-Zyklus durchliefen 0.1 bis 0.5% der phosphorylierten Glucose. Phagozytose erhöhte diesen Anteil auf das 2- bis 3-fache. Zwischen normalen und diabetischen Zellen fanden sich dabei keine Unterschiede. — 4. Milchsäure stellte das Hauptendprodukt des Glucose-stoffwechsels dar. Die Verteilung des ¹⁴C aus der Glucose spiegelte nicht nur die Einwirkungen des oxydativen und des nicht-oxydativen Anteils des Pentose-Phosphat-Zyklus wider, sondern deutete auch auf die Beteiligung eines symmetrischen C₃ Zwischenproduktes, nämlich Dihydroxyaceton, beim Stoffwechsel eines Teils des Glucose-Kohlenstoffs hin. — 5. Nur sehr kleine Mengen des ¹⁴C aus Glucose fanden sich in Aminosäuren, nicht mit Äther extrahierbaren Stoffwechsel-Säuren und Lipiden. Hingegen ließen sich beträchtliche Anteile des ¹⁴C in einem Stoffgemisch (A) nachweisen, von dem ein Teil als Glycerin-Tribenzoat präzipitierte, und in einem Gemisch (B), das wahrscheinlich Eiweißcharakter besitzt. Die Einlagerung von ¹⁴C in das Gemisch A durch normale Leukozyten nahm während der Phagozytose ab und war unbedeutend bei diabetischen Zellen. Der Einbau von ¹⁴C aus [1-¹⁴C]- und [2-¹⁴C]-, nicht aber aus [6-¹⁴C]-Glucose in das Gemisch B war bei normalen Zellen während der Phagozytose erheblich gesteigert, jedoch nicht bei diabetischen Zellen.

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Le métabolisme du glucose [1-¹⁴C]-, [2-¹⁴C]-[3,4-¹⁴C]- et [6-¹⁴C]- dans des leucocytes normaux et diabétiques polynucléaires et au cours de la phagocytose

Résumé 1. Des leucocytes polynucléaires de sujets normaux et diabétiques et de rats normaux et rendus diabétiques par l'alloxane ont été incubés avec du ¹⁴C-glucose et mis à phagocyter. — 2. La majeure production de ¹⁴CO₂ venait du cycle des pentoses. Approximativement un tiers du ¹⁴CO₂ était formé par la decarboxylation du pyruvate, tandis que l'activité du cycle de Krebs était minime. — 3. Le cycle des pentoses métabolisait 0.1–0.5% du glucose phosphprylé, et la phagocytose augmentait 2–3 fois cette fraction. On n'a trouvé aucune différence entre les cellules normales et les cellules de diabétiques. — 4. Le produit final principal du métabolisme du glucose était l'acide lactique. La dispersion du ¹⁴C du glucose n'était pas compatible avec le seul fonctionnement des parties oxydatives et non-oxydatives du cycle des pentoses, mais indiquait la participation d'un intermédiaire symétrique en C₃, c-à-d la dihydroxyacétone, dans le métabolisme d'une partie des carbones du glucose. — 5. De très petites quantités de ¹⁴C du glucose ont été trouvées dans les acides aminés, les acides métaboliques non-extractibles par l'éther et dans les lipides. Cependant on a trouvé des quantités appréciables de ¹⁴C dans un composé (A) dont une partie précipitait comme tribenzoate de glycérol, et un composé (B) qui était probablement de nature protéique. L'incorporation de ¹⁴C dans le composé A de leucocytes normaux diminuait au cours de la phatocytose et était insignifiante dans les cellules de diabétiques humains. — L'incorporation de ¹⁴C du glucose [1-¹⁴C]- et [2-¹⁴C]-, mais non du glucose [6-¹⁴C] dans le composé B était fortement stimulée dans les cellules normales au cours de la phagocytose, mais pas autant dans les leucocytes de diabétiques.

     

Association study of CD14 polymorphism with myocardial infarction in a Japanese population

There have been many studies investigating the association between gene polymorphisms and coronary artery disease (CAD) including myocardial infarction (MI), and some studies have shown that certain gene polymorphisms are associated with CAD/MI. However, the results of the association have sometimes been controversial. The reason may be that the contribution of genetic risk factors to CAD/MI varies depending on the ethnic, environmental, and habitual backgrounds, and differs between males and females. In this study, we analyzed 17 polymorphisms in 12 candidate genes for MI in 136 patients and 200 to 235 controls, and found that there is a significant association of MI with the polymorphisms in the genes for E-selectin and CD14 receptor. To further explore the association, we investigated the C-260 T polymorphism in the promoter region of the CD14 gene in 502 MI patients and 527 control subjects. The genotype distributions of the CD14 polymorphism were as follows: patients; T/T 32.5%, C/T 48.2%, C/C 19.3%, and controls; T/T 25.4%, C/T 52.8%, C/C 21.8%. The frequencies of the T/T homozygotes were significantly higher in the patients (OR = 1.41, P = 0.013) than in the control group, confirming the association of CD14 polymorphism with MI in Japanese. Stratification analyses further demonstrated that the association was more prominent in females and in patients with a relatively low body mass index, suggesting that the contribution of the CD14-linked genetic risk to MI differs with respect to gender and habitual background.     

CD14 -550 C/T, which is related to the serum level of soluble CD14, is associated with the development of respiratory syncytial virus bronchiolitis in the Japanese population

BACKGROUND: The contribution that genetic polymorphisms of Toll-like receptor (TLR) 4 and of CD14--both of which recognize respiratory syncytial virus (RSV) in the innate immune response--make to RSV bronchiolitis in the Japanese population has not yet been clarified. METHODS: This study genotyped 2 TLR4 mutations, Asp299Gly and Thr399Ile, and 2 single-nucleotide polymorphisms (SNPs) of CD14, -550 C/T and -159 C/T, in 54 children with RSV bronchiolitis and in 203 control subjects. CD14 SNPs and the serum level of soluble CD14 (sCD14) also were examined in 67 cord-blood specimens and in serum samples from 73 6-year-old children. RESULTS: No TLR4 mutations were found. The frequencies of both the CC genotype and the C allele of CD14 -550 C/T were significantly higher in children with RSV bronchiolitis than in the control subjects. The serum level of sCD14 was significantly higher in children with the CC genotype of CD14 -550 C/T than in those with the CT and TT genotypes. CONCLUSIONS: CD14 -550 C/T, which is related to the serum level of sCD14, is associated with the development of RSV bronchiolitis in the Japanese population. This study's results indicate that, in different ethnic groups, different genetic factors contribute to the development of RSV bronchiolitis.     

Expression of ADAMTS-2, -3, -13, and -14 in culprit coronary lesions in patients with acute myocardial infarction or stable angina

ADAMTS (a disintegrin and metalloproteinase with thrombospondin type 1 motifs) proteases are emerging as key participants in the pathogenesis of vascular diseases. We studied the expression of ADAMTS-2, -3, -4 and -14 in the culprit plaques from patients presenting with acute myocardial infarction (AMI) versus stable angina. Tissue samples were gathered from 52 patients with AMI (n = 35) or stable angina (n = 17) who underwent directional coronary atherectomy. The specimens were stained with hematoxylin-eosin and analyzed immunohistochemically using antibodies specific to ADAMTS-2, -3, -13 and -14, and markers for endothelial cells, macrophages, and smooth muscle cells. Baseline characteristics of the groups were mostly similar. The proportion of smooth muscle α-actin-immunopositive area was smaller in the AMI group than in the stable angina group, but the areas immunopositive for CD31 or CD68 were higher in the AMI group. The relative areas immunopositive for ADAMTS-2, -3, and -13 in AMI were significantly larger than those in stable angina. However, the proportion of areas immunopositive for ADAMTS-14 did not differ between the two groups. Areas that stained for ADAMTS-2, -3, -13, and -14 largely overlapped with those positive for CD31 or CD68. The areas immunopositive for ADAMTS proteases were significantly correlated with CD31- or CD68-immunostained areas. In conclusions, ADAMTS-2, -3, and -13 expression, but not that of ADAMTS-14, are increased in plaques causing AMI compared those associated with stable angina. These results support a role for these enzymes in the pathogenesis of AMI.