Identification of natural antibodies to interleukin-18 in the sera of normal humans and three nonhuman primate species

Natural antibodies to cytokines can be found in the sera of normal healthy individuals in the absence of specific immunostimulation. However, the function, impact, and purpose of natural antibody development have yet to be fully elucidated. Interleukin (IL)-18 is a cytokine that exerts proinflammatory activities and induces natural killer (NK) cell activity. Recombinant human IL-18 (rHuIL-18) is currently in development as a cancer immunotherapy. In this study, the presence of natural antibodies to IL-18 in the sera of normal humans and three nonhuman primate species was evaluated by electrochemiluminescence immunoassay (ECLIA). Of the human sera tested, 6 of 47 samples were positive for natural antibodies to IL-18. Of the nonhuman primate sera tested, 22 of 80 cynomolgus monkey samples, 4 of 31 rhesus monkey samples, and 2 of 20 chimpanzee samples were positive for natural antibodies to IL-18. Natural anti-IL-18 antibodies were neutralizing in 5 of 22 cynomolgus and 2 of 4 rhesus sera. None of the chimpanzee or human sera were able to neutralize IL-18 induction of interferon (IFN)-gamma in vitro. In vivo activity of rHuIL-18 was compared in IL-18 natural antibody-positive and -negative cynomolgus monkeys. The presence of natural antibodies to IL-18 did not alter rHuIL-18 systemic exposure levels, induction of neopterin, or induction of treatment-induced antibodies following intravenous administration of rHuIL-18. In conclusion, our data indicate that, as has been found with other cytokines, natural anti-IL-18 antibodies are relatively common. Moreover, natural anti-IL-18 antibodies do not appear to influence rHuIL-18 activity in vivo and are not predictive of a heightened immune response, suggesting that natural anti-IL-18 antibodies do not impact IL-18 therapy. Finally, our data suggest that the ability to detect natural anti-cytokine antibodies may be a useful measure of the adequacy of an assay for deployment in clinical trials.     

电化学发光法测定红细胞叶酸水平的性能评价

目的 评价电化学发光法测定红细胞叶酸浓度的方法.方法 通过重复性试验、回收试验、干扰试验、线性试验和方法学比对对本方法进行评价,并建立本方法的参考范围;在此基础上对30例补充叶酸60天以上的孕妇的红细胞叶酸水平进行测定.结果 本方法的批内和批间的变异系数（CV）分别为2.57％和3.55％,回收率为98.0％,线性范围为40.2 ～ 1174.82ng/mL,甘油三酯（148mg/dL）和胆红素（0.6mg/dL）不会对本实验造成影响.本实验室红细胞叶酸值的参考范围是111.33～603.75ng/mL.本法测得的红细胞叶酸结果与放射免疫法结果比较相关性良好（r=0.998）,两种方法测定结果差异无显著性.30例补充叶酸60天以上的孕妇的红细胞叶酸水平为911.23±198.19ng/mL,与对照组（423.57±135.78 ng/mL）相比差异有统计学意义（P＜0.01）.结论 电化学发光法能够准确、快速的测定红细胞叶酸浓度,且该法重复性好,适合应用于临床检测.     

Ionic liquid-based hollow fiber-supported liquid-phase microextraction enhanced electrically for the determination of neutral red

A method of ionic liquid-based hollow fiber liquid-phase microextraction enhanced electrically was successfully developed and applied to the extraction and determination of neutral red (NR) dye, which was selected as the model analyte. A room temperature ionic liquid, 1-octyl-3-methylimidazolium hexafluorophosphate ([C₈mim][PF₆]), was placed in the pores of a polytetrafluoethylene hollow fiber, which acts as a liquid membrane and the acceptor solution. The extraction parameters affecting the enrichment factor of NR, such as pH, extraction time, elution time, stirring rate, and the voltage were optimized. In addition, UV–Visible (UV–Vis) or electrochemiluminescence spectra were also determined. The extraction rate and capacity of NR could be improved significantly by cathodic polarization. Under the optimized extraction conditions (organic liquid microextraction phase [C₈mim][PF₆], pH 7, stirring rate 300 rpm, extraction time 20 minutes, ultrasonic-assisted elution time 3 minutes, voltage −70 V), the detection limit of 0.38 μg/L and linear correlation coefficient of r > 0.99 were obtained. The established method was successfully applied to the analysis of three soft drink samples, which were spiked with NR standards at the concentrations of 0.1, 1.0, and 5.0 mg/L, and satisfactory results were obtained.     

永康地区正常妊娠妇女甲状腺功能指标参考值的研究

目的建立永康市正常孕妇妊娠各期甲状腺功能指标的参考值。方法收集2017年6月-12月在永康市妇幼保健院产检妊娠妇女为研究对象,经过筛选共纳入449例。采用全自动电化学发光免疫分析方法进行甲状腺功能7个项目的测定,并依测定数据统计分析得到各个项目的双侧95%医学参考值范围。结果研究获得本院正常妊娠妇女各孕期甲状腺激素的参考值如下:早孕期,TSH:0.04μIU/ml^4.14μIU/ml,FT4:1.01 ng/dl^1.78 ng/dl,FT3:2.52 pg/ml^4.31 pg/ml;中孕期,TSH:0.02μIU/ml^4.77μIU/ml,FT4:0.85 ng/dl^1.71 ng/dl,FT3:2.34 pg/ml^4.39 pg/ml;晚孕期,TSH:0.24μIU/ml^5.63μIU/ml,FT4:0.74 ng/dl^1.30 ng/dl,FT3:2.11 pg/ml^3.58 pg/ml。年龄对妊娠妇女甲状腺功能影响并不大。根据美国甲状腺协会推荐标准和本研究的参考值诊断本院妊娠妇女亚临床甲状腺功能减退的患病率分别为17.59%和2.23%。结论甲状腺激素在妊娠各期之间的差异有统计学意义,不同地区妊娠妇女甲状腺激素参考值也存在差异,建立适合本地区的特异性甲状腺激素参考值有助于妊娠期甲状腺疾病的诊疗。     

Comparison between a new multiplex electrochemiluminescence assay and the WHO reference enzyme-linked immunosorbent assay to measure serum antibodies against pneumococcal serotype-specific polysaccharides

Background

Two electrochemiluminescence (ECL) assays were developed which, together, can simultaneously measure serum antibodies against pneumococcal capsular polysaccharides (PnPS) for 17 serotypes. The assays were validated for the 13 PnPS included in the 13-valent pneumococcal conjugate vaccine (PCV13). As recommended by the World Health Organization (WHO), we compared the ECL assays with the WHO reference enzyme-linked immunosorbent assay (ELISA) and derived a threshold corresponding to the 0.35?µg/mL threshold established for the WHO reference ELISA for the non-inferiority comparison and licensure of new PCVs against invasive pneumococcal disease.

联合检测CA125、CYFRA21-1、NSE和CEA对肺癌诊断价值的探讨

目的 探讨糖类抗原125（CA125）、细胞角蛋白21-1（CYFRA21-1）、神经元特异性烯醇化酶（NSE）、癌胚抗原（CEA）联合检测对肺癌的诊断价值.方法 应用检测灵敏度高和线性范围宽的电化学发光技术对68例肺癌、41例肺良性疾病及33例健康体检者血清进行检测.结果 肺癌组血清CA125、CYFRA21-1、NSE和CEA含量均显著高于健康对照组和肺良性疾病组（P〈0.01）.单项肿瘤指标对肺癌检测敏感性分别为73.5%、75.0%、79.4%、82.4%,联合检测可提高检测敏感性达97.1%;如以任何三项以上阳性作为肺癌的诊断依据,则敏感性为75.0%,而对照组无1例阳性.结论 CA125、CYFRA21-1、NSE和CEA联合检测可显著提高肺癌诊断的敏感性,在肺癌的早期诊断方面可为临床提供有价值的资料.     

标本溶血对电化学发光免疫法结果的影响

目的探讨标本不同程度溶血对电化学发光免疫法(ECLIA)测定结果的影响。方法将血液标本人为干预成不同程度的溶血标本,使用ECLIA检测各组标本中的甲状腺功能、肿瘤标志物、内分泌激素等共28项指标的浓度并做分析。结果标本溶血可使胰岛素(INS)浓度降低,使叶酸(FA)、神经元特异性烯醇化酶(NSE)浓度升高,重度溶血有使铁蛋白测定结果偏高的趋势,但差异无统计学意义,其余指标测定结果不受溶血的影响。结论除某些项目外大部分溶血标本的ECLIA测定结果没有受到影响。测定INS、FA、NSE时应绝对避免使用溶血标本,以确保结果准确。     

广州番禺地区儿童总钙和25-羟基维生素 D检测结果分析

目的：统计广州市番禺地区儿童总钙和25-羟基维生素D[25（OH）D]水平并进行分析,为以后该地区儿童保健提供参考依据。方法使用原子吸收光谱仪和电化学发光仪检测1902名儿童总钙和25（ OH） D水平。将受检儿童按年龄不同分为5个年龄组：婴儿组（1～12个月,817名）、幼儿组（1～2岁,599名）、学龄前组（3～6岁,291名）、学龄期组（7～11岁,167名）、青春发育期组（12～18岁,28名）。在按年龄分组的基础上再按性别分成男、女两组。对各组总钙和25（ OH） D水平进行统计分析。结果广州番禺地区儿童总钙水平为（1．72±0．19）mmol／L,25（OH）D 水平为（39．12±11．26）ng／mL,两者均随年龄的增长呈下降趋势。按年龄分组比较显示婴儿组总钙及25（OH）D水平均明显高于其余4组（P均＜0．05）;幼儿组总钙水平明显高于学龄前组及学龄期组（P均＜0．05）,25（OH）D水平明显高于学龄前组、学龄期组及青春发育期组;学龄前组总钙及25（OH）D水平均高于学龄期组（P均＜0．05）。男、女性儿童总钙和25（OH）D水平差异均无统计学意义（P均＞0．05）。总钙和25（OH）D呈正相关[相关系数（r）＝0．221,P＜0．05,线性方程为Y总钙＝0．004X25（OH）D ＋1．573]。结论番禺地区儿童血液总钙和25（ OH） D水平偏低。建议学校和家长能督促儿童及时、合理的补钙。     

电化学发光免疫分析检测乙型病毒性肝炎患者血清 AngⅡ与 HA 的相关性研究

目的：通过电化学发光免疫分析检测,探讨乙型病毒性肝炎（以下简称乙肝）患者的血清血管紧张素Ⅱ（AngⅡ）与透明质酸酶（HA）之间的关系。方法选取2012年1月至2014年1月高密市人民医院检验科收治的乙肝患者100例,其中急性乙肝患者50例（急性组）,慢性乙肝患者50例（慢性组）,选择同期在我院体检的健康人群50例作为对照组。采用速率法检测血清 AngⅡ水平,电化学发光法检测 HA 水平,对三组的结果进行统计分析。结果慢性组血清 AngⅡ和 HA 水平明显高于急性组和对照组,急性组明显高于对照组,差异有统计学意义（t =1.306、2.839、1.804,P 均〈0.05）;Pearson 相关系数分析显示：血清 AngⅡ和 HA 水平呈正相关（r =0.703,P 〈0.05）。结论乙肝患者血清 AngⅡ和 HA 水平关系密切,两者均可作为判断肝纤维化程度的重要指标。     

电化学发光法在检测孕产妇乙型肝炎病毒中的应用

目的了解电化学发光法在检测孕产妇乙型肝炎病毒中的应用价值,从而有助于正确指导乙肝孕产妇产后的喂养方式。方法用ELISA法和电化学发光法平行测定孕产妇血清乙型肝炎病毒表面抗原,并对ELISA法检测为阴性而电化学发光法为低浓度的标本,用荧光定量PCR测定病毒DNA。结果电化学发光法对表面抗原的检出率明显高于ELISA法,ELISA法检测为阴性而电化学发光法为低浓度的标本,病毒DNA为阳性。结论电化学发光法检测血清表面抗原的灵敏度高于ELISA法,可以更好的指导孕产妇产后的喂养方式。