破伤风类毒素的发展历史、现状和新型破伤风疫苗的研究进展

简要回顾了破伤风类毒素主要的发展历程;概述了目前国内外精制破伤风类毒素的两种生产工艺各自的优缺点,并且肯定了吸附精制破伤风类毒素几十年实际应用的效果,同时指出了在大规模人群接种时可能出现为数不多的各种不良反应,以及引起不良反应的可能的原因。对国内外破伤风疫苗研究进展和主要发展方向,如亚单位疫苗和新型破伤风类毒素的研究概况和前景也作了介绍。     

一种提高破伤风类毒素马匹免疫成功率的方法

采用抗原加倍方法用于成率率低于705群的巴匹免疫,结果显示,进一步免疫成功的马匹明显多于常规免疫（P＜0．05）,免疫效价和死亡率无明显变化（P＞0．05）。     

破伤风类毒素不良反应述评

本文简略回顾了破伤风类毒素接种后不良反应的历史,介绍了反应（主要是超敏反应）的类型,讨论了发生反应的原因,提出了减少反应的措施。     

用体外免疫的人淋巴细胞建立抗破伤风类毒素的...

Human tonsil cells in vitro immunized with tetanus toxoid were fused with human-mouse heteromyeloma line RF to generate human-mouse hybridomas. Hybridoma 891112-50 was cloned and 2 subclones (891112-50-3 and -4) were obtained. The secreted antibodies from the subclones were antigen specific, since they did not cross react with three irrelevant antigens (OVA, TCS and F gamma G). The hybridomas were quite stable. After 13 passages in tissue culture flasks, they still retained their antibody secreting ability. From flow cytometry analysis the subclone 50-3 was more stable than the subclone 50-4. The human immunoglobulin contained in supernatant collected during regular passages was equivalent to 69.6 micrograms/ml.     

破伤风类毒素抗体酶联双抗原夹心法定量检测试剂的研制及评价

研制破伤风类毒素抗体酶联双抗原夹心法定量检测试剂,用于破伤风免疫血浆抗体效价检测。以精制破伤风类毒素经Sephacryl S-300柱层析纯化后作为包被抗原,用辣根过氧化物酶以改良过碘酸钠法标记精制破伤风类毒素作为酶标记抗原,以破伤风人免疫球蛋白国家标准品采用小鼠中和试验法标定试剂盒定量标准品,制备双抗原夹心法定量检测试剂;进行试剂盒检测范围、特异性、重复性、精密度及稳定性考核,并与小鼠中和试验法、琼脂双扩散法及国外破伤风类毒素抗体酶联试剂盒进行比较。结果显示,试剂盒的检测范围为10~150mIU/ml,灵敏度为10mIU/ml,线性好(r>0.996),板内孔间变异度小(CV<8%),特异性强(100%),重复性好(CV<13%),于37℃放置6天测定结果无明显差异,与小鼠中和试验法、英国Biding Site酶联试剂有良好的一致性。试验证明所研制的试剂盒适用于破伤风免疫血浆中的破伤风抗体效价定量检测。     

14型肺炎球菌荚膜多糖-破伤风类毒素结合疫苗的研制

将14型肺炎球菌的荚膜多糖(PS)与破伤风类毒素(TT)通过化学方法结合,制备成多糖-蛋白结合疫苗(PS14-TT)。用该结合疫苗免疫小鼠,在小鼠体内产生了高滴度的PS-IgG抗体和TT-IgG抗体,且再次注射后有加强应答效应,表明制备的结合疫苗保留了完好的抗原性,具有胸腺依赖性抗原的特性。     

双佐剂包被的破伤风类毒素马匹免疫试验

为提高破伤风免疫马匹的血浆抗体效价,应用不同佐剂配制TT抗原,进行马匹超免疫比较研究;采用FIA和植物油双佐剂包被与单佐剂包被的TT抗原,注射马匹进行超免疫,比较三组血浆的效价;结果显示,双佐剂抗原较单佐剂的免疫效果好,但可能对马匹刺激较强,有待调整注射剂量和免疫程序。     

关于吸附精制百日咳菌苗、白喉、破伤风类毒素混合制剂的现况

本文介绍了现行吸附百日咳菌苗、白喉、破伤风类毒素混合制剂的现况和新一代吸附精制百日咳菌苗、白喉、破伤风类毒素混合制剂人体接种反应和血清学效果观察的结果。     

吸附无细胞百白破(DPTa)的稳定性试验

采用半综合液体培养基（含０．０５％ＭｅβＣＤ）,大罐培养百日咳Ⅰ相ＣＳ菌制备的吸附无细胞百日咳菌苗、白喉、破伤风类毒素混合制剂（ＤＰＴａ）置４－８℃分别保存一年、保存两年,测三种抗原成分的效力及毒性,以及将该制剂于３７℃分别放置三周、三个月,测定有无毒性逆转。结果表明,该制剂质量稳定,无毒性逆转     

Collaborative study for the calibration of a replacement International Standard for Tetanus Toxoid Adsorbed

We present the results of a collaborative study for the establishment of a replacement International Standard (IS) for Tetanus Toxoid Adsorbed. Two candidate preparations were included in the study, one of which was established as the 4th IS for Tetanus Toxoid Adsorbed at the WHO Expert Committee on Biological Standardization meeting in October 2010. This preparation was found to have a unitage of 490 IU/ampoule, based on calibration in guinea pig challenge assays. Results from mouse challenge assays suggest that the relative performance of two candidate preparations may differ significantly between guinea pigs and mice. The authors note that the number of laboratories that performed guinea pig challenge assays, which are used to calibrate and assign IU, is much lower than in previous collaborative studies and this may have implications for calibration of replacement standards in the future. The issue of assigning separate units to the IS for guinea pig and mouse assays is discussed. The study also assessed performance of the replacement standard in serological assays which are used as alternative procedures to challenge assays for tetanus potency testing. Results suggest that the replacement standard is suitable for use as the reference vaccine in serological assays.     

The Immunopurification of Tetanus Toxoid

S ummary . The isolation of tetanus toxoid by immunosorption chromatography is described. The immunosorbent, a polymerized horse antitoxin, was used repeatedly with little loss in capacity and recoveries of toxoid of > 90% were achieved. In terms of specific antigenic activity, immunopurified tetanus toxoid is purer than that purified by conventional procedures and is at least as immunogenic.     

Determination of Tetanus Toxin and Toxoid by ELISA Using Monoclonal Antibodies

The procedure for obtaining monoclonal antibodies TT-1, TT-2, and TT-3 against tetanus toxin/toxoid is described. It is shown that the commercial DTP vaccine and tetanus toxoid conjugated with a low-molecular-weight hapten can both be used as immunogens. Monoclonal antibodies TT-1 and TT-2 neutralized tetanus toxin in vivo. The monoclonal antibodies obtained were used to design and compare several schemes of quantitative determination of tetanus toxoid and toxin by ELISA. A more sensitive competitive ELISA allowed the detection of as much as 0.01 EC/ml toxoid and 50 LD₅₀/ml toxin.