物化与免疫化学技术预测白喉类毒素质量

制备白喉类毒素疫苗最关键的步骤是甲醛脱毒。白喉类毒素(DTx)的脱毒过程通过毒素发生的部分未知化学改变来实现。因此白喉疫苗很难完全确定特性,类毒素质量也只能用常规效力与安全实验测定。本文描述了选择物理化学与免疫化学试验以替代原有体内试验监测白喉类毒素疫苗产品质量的可能性。     

不同粒径氢氧化铝佐剂对白喉类毒素免疫效果的影响

研究不同粒径的氢氧化铝[Al(OH)3]对白喉类毒素的吸附效果,用于指导疫苗生产,提高疫苗质量。用透射电镜测得的不同粒径的Al(OH)3分别吸附白喉类毒素及用絮状单位测定法和疫苗效价测定法对其吸附效果作比较,并经统计学处理。试验结果显示,粒径为210nm的Al(OH)3对白喉类毒素吸附效果及对白喉疫苗效力的免疫增强作用明显好于粒径600nm的Al(OH)3。实验证实,Al(OH)3佐剂的粒径大小与白喉类毒素的吸附效果及疫苗的免疫原性密切相关。     

关于吸附精制百日咳菌苗、白喉、破伤风类毒素混合制剂配方的实验报告

本文对吸附精制百白破混合制剂的不同配方进行了实验,结果表明,新一代吸附精制百白破混合制剂最佳配方为：精制百日咳菌苗１８μｇＰＮ／ｍｌ、精制白喉类毒素为３０Ｌｆ／ｍｌ、精制破伤风类毒素为１０Ｌｆ／ｍｌ。由该配方组成的吸附精制百白破混合制剂,其中百日咳菌苗的毒性试验ＢＷＤＵ／ｍｌ、ＬＰＵ／ｍｌ、ＨＳＵ／ｍｌ三种指标均符合制检规程要求。其效力单位（ＩＵ／ｍｌ）超过规程要求一倍以上,精制白喉和破伤风类毒素的安全试验均符合规程要求,白类效力试验≥８０－１００％,破类效力试验≥０．５－４．５ＩＵ／ｍｌ。上述结果说明本文提出的配方作为新一代精制百白破混合制剂的配方是适宜和实用的。     

精制白喉毒素加β—丙氨酸后脱毒效果观察

精制白喉毒素加０．０２Ｍβ—丙氨酸,再加甲醛溶液经适当的时间解毒,即可转化为完全类毒化且无毒性逆转的精制白喉类毒素。此精白类的脱毒试验、毒性逆转试验、安全试验及效力试验均符合《中国生物制品规程》要求。在脱毒过程中絮状单位的损失明显低于单纯甲醛脱毒者,纯度亦相应得到了提高。     

吸附精制百日咳菌苗毒性逆转试验报告

吸附精制百日咳菌苗毒性逆转试验报告史天绪,陈爱荣,秦力,何长民（卫生部兰州生物制品研究所,兰州７３００４６）目前已知,吸附精制百日咳菌苗或吸附精制百日咳、白喉、破伤风类毒素混合制剂（简称ＡＤＴＰ）和吸附精制百日咳、白喉类毒素混合制剂（简称ＡＤＰ）,如...     

朝圣者接种四价脑膜炎球菌TT结合疫苗前后施用Tdap对脑膜炎球菌多糖抗体应答的影响

<正>朝圣者易受多重严重感染,需要接种多种疫苗。多糖蛋白结合疫苗含有破伤风类毒素(TT)、白喉类毒素或白喉类毒素突变体(CRM197)等载体蛋白。这些载体蛋白可能会与其他结合疫苗或含有破伤风或白喉的联合疫苗在同时或连续给药时发生相互作用。作者研究了在2015年参加朝圣前的澳大利亚成年朝圣者中单独和联合使用破伤风/白喉/无细胞百日咳(Tdap)疫苗与四价脑膜炎球菌TT结合疫苗(MCV4)与13价肺炎球菌CRM197结合疫苗(PCV13)联合使用时的免疫相互作用。将每个参     

4种疫苗脂质体合成方法的研究

目前在国内将脂质体用于疫苗的领域尚未见报道。我们摸索了不同的脂质体合成技术,制备合适的脂质体包被甲肝减毒活疫苗,重组乙肝疫苗、白喉类毒素和破伤风类毒素,开发脂质体疫苗的合成途径,以根据不同疫苗的特点和免疫途径生产相应的脂质体疫苗。1材料与方法1．1材料磷脂酸胆碱（Phoophatidvlserline,PC）,磷脂酸丝氨酸（Phesphatidylserline,PS）,Choesteral（CH）均为西格玛公司产品。甲肝减毒活疫苗,CHO细胞重组乙肝疫苗,破伤风类毒素,白喉类毒素为我所生产的半成品,各项检定指标均合格。1．2方法脂质体粒径和粒度分布…     

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。     

10价肺炎球菌性流感嗜血杆菌蛋白D结合疫苗的有效性

背景：设计了芬兰侵袭性肺炎球菌性疾病疫苗临床试验,以期评价肺炎球菌性疫苗的有效性,这种疫苗含有10种与流感嗜血杆菌蛋白D、破伤风类毒素及白喉类毒素作为载体蛋白相结合的血清型特异的多糖（PHiD—CV10）。     

鼠疫F1抗原和V抗原双组分候选疫苗的免疫学评价

目的通过由提取的鼠疫F1抗原和重组鼠疫V(rV)抗原组成的鼠疫候选疫苗免疫豚鼠,对其免疫效果进行评价。方法将豚鼠随机分成5个试验组,在免疫后不同时间点采血进行抗体检测、MTT法淋巴细胞增殖试验以及皮肤迟发型超敏反应(DTH)检测。结果抗体检测结果显示,双组分鼠疫候选疫苗能诱导较强的体液免疫应答;MTT细胞增殖结果显示,脾脏淋巴细胞特异性增殖不明显;中剂量组、高剂量组针对F1和rV抗原DTH阳转率均为100%。结论双组分鼠疫候选疫苗能诱导豚鼠体液免疫和细胞免疫应答,该疫苗有希望成为我国新一代鼠疫疫苗。     

In vitro induction of a diphtheria toxoid specific antibody response in human peripheral blood lymphocytes cultivated in the presence of diphtheria toxoid

In comparison with the presently used potency test for diphtheria vaccine, in vitro examination of the immunogenicity of the vaccine would have great advantages. For this reason in vitro induction of diphtheria toxoid specific antibody synthesis in human peripheral blood lymphocytes cultivated in the presence of diphtheria toxoid was investigated. The results showed that a dose dependent synthesis of diphtheria antibody was induced by adsorbed diphtheria toxoid and combined vaccines containing the diphtheria toxoid component. Plain diphtheria toxoid appeared to be less immunogenic in comparison with adsorbed toxoid. There is some indication that the pertussis component had a stimulating effect on the diphtheria antibody synthesis. In conclusion, these results are promising for in vitro examination of the immunogenicity of diphtheria vaccines. The model will be validated for the routine control of diphtheria vaccine.     

Specific depression of delayed hypersensitivity to purified proteins, with relation to production of circulating antibody

In guinea pigs sensitized in the footpads with a purified protein, such as hen egg albumin or diphtheria toxoid, in incomplete Freund's adjuvant, delayed hypersensitivity precedes the appearance of circulating antibody. This expression of delayed hypersensitivity by skin-test declines sharply the day before circulating antibody is detected. Adoptive transfer of spleen and peritoneal exudate cells from guinea pigs showing this decline suppresses the expression of delayed hypersensitivity in already sensitized recipients. This suppression of delayed hypersensitivity is immunologically specific. The intensity of this suppression does not correlate directly with the dose of sensitizing antigen, nor does it depend directly on the amount of circulating antibody.     

Influence of genetic and phenotypical factors on the efficiency of the vaccination of young children against diphtheria and measles

The child's sex was shown to influence the character of antibody formation only after immunization against diphtheria with live measles vaccine: girls exhibited stronger reaction to vaccination than boys. Children of different gender were found to have characteristic HLA DR markers of humoral immune response to diphtheria toxoid and measles vaccine. HLA DR7 proved to be the marker of low production of antibodies to diphtheria toxoid and measles vaccine in boys.     

Simultaneous Immunization with B.C.G., Diphtheria-tetanus,and Oral Poliomyelitis Vaccines in Children Aged 13-14

The simultaneous administration of B.C.G. vaccine, diphtheria-tetanus toxoid aluminium hydroxide adsorbed vaccine, and oral poliovaccine was studied in 628 children aged 13-14 years between 1966 and 1969 in Newham, London. The efficacy of these vaccines was unaffected by administering them at the same time; routine simultaneous administration is considered justified when organizational difficulties prevent the attainment of high immunization rates with the vaccines given separately. No adverse reactions to B.C.G. or oral poliomyelitis vaccines took place, but 8% of children had moderately severe local reactions after diphtheria-tetanus aluminium hydroxide adsorbed vaccine, which were attributed to diphtheria toxoid.Serological studies showed the need for immunization against diphtheria, tetanus, and poliomyelitis at 13-14 years of age. Because of the adverse reactions to diphtheria toxoid, however, simultaneous administration of tetanus toxoid aluminium hydroxide adsorbed, oral poliomyelitis, and B.C.G. vaccines only is recommended at present.An “adult type” diphtheria-tetanus toxoid might overcome the problem of reactions, though in two to three years'' time most children aged 13-14 years will have received diphtheria-tetanus-pertussis vaccine in infancy and reinforcement might then be accomplished by a small intradermal dose of the currently available fluid diphtheria-tetanus vaccine.Continued serological studies of diphtheria and tetanus antitoxins and polio antibody are necessary to determine the future need for reinforcement of immunity; such studies should become an essential part of the surveillance of the community immunization programme.     

Comparative quantitation for the protein content of diphtheria and tetanus toxoids by DC protein assay and Kjeldahl method.

DPT, a combination vaccine against diphtheria, tetanus and pertussis is available since many years and still continued in the national immunisation schedule of many countries. Although highly potent, reactions to DPT vaccine are well known, mainly attributed to the factors like Pertussis component, aluminum adjuvant and lower purity of tetanus and diphtheria toxoids. The latter most important aspect has become a matter of concern, specially for the preparation of next generation combination vaccines with more number of antigens in combination with DPT.Purity of toxoid is expressed as Lf (Limes flocculation) per mg of protein nitrogen. The Kjeldahl method (KM) of protein nitrogen estimation suggested by WHO and British Pharmacopoeia is time consuming and less specific. Need has been felt to explore an alternative method which is quick and more specific for toxoid protein determination. DC (detergent compatible) protein assay, an improved Lowry's method, has been found to be much more advantageous than Kjeldahl method.     

Preparative Procedure for the Purification of Toxoids by Gel Filtration

Sephadex gel filtration can be employed as a preparative procedure for the purification of both tetanus and diphtheria toxoids. A toxoid purification sequence is described in the text. By utilizing the described methods and columns, up to 100,000 human doses of diphtheria toxoid could be processed in a single operation. The method has given an 80% yield of diphtheria toxoid with a purity of 1,900 Lf per mg of N. The analysis of the material by immunodiffusion tests showed that a marked increase in purity was achieved. Antigenicity tests demonstrated that there was no significant difference in antigenic potency between the parent toxoid and its purified fraction. Factors limiting the effective separation of tetanus toxoid by gel filtration are discussed. The construction of the columns used is described in detail, as well as packing procedures and column characteristics such as bed volume, void volume, sample size, and flow rate.     

The humoral immunity against diphtheria

Results of the conducted study showed that naturally acquired antibacterial and postvaccinal antitoxic antibodies against diphtheria were found in human blood sera. Challenge of ADT-M toxoid to adults resulted in production of antitoxic as well as antibacterial antibodies in high concentrations. In response to challenge of ADT-M toxoid simultaneously with bacterial vaccine against diphtheria Codivac both antibacterial and antitoxic antibodies were synthesized in blood on optimal physiologic levels. This study revealed dynamics of some specific characteristics of humoral immune response after challenge of two different vaccines against diphtheria--ADT-M toxoid and Codivac vaccine.     

Optimization of diphtheria toxin production by <Emphasis Type="Italic">Corynebacterium diphtheriae</Emphasis> using a casein-based medium in a fermenter

The Td-based combined vaccine contains only small amounts of the diphtheria toxoid antigen. However, a high level of purity is necessary for this antigen. The diphtheria toxin is produced by growing Corynebacterium diphtheriae in a semisynthetic, casein-based medium in a fermenter. In order to obtain a highly pure diphtheria toxoid, the optimal conditions to express the toxin at 300 Lf/mL in a fermenter culture were determined. When C. diphtheriae was cultivated in a fermenter and a high concentration of toxin was obtained, specific patterns for the pH and dissolved oxygen levels identified. Overall, the fermenter cultivation process was divided into four stages according to variations in the pH. A specific range of K _La in the fermenter (0.0092 ~ 0.0093/sec) was required to produce high level expression of diphtheria toxin. The amount of toxin expression varied significantly according to culture conditions. Agitation and aeration in the fermenter affected toxin expression, even when the optimal K _La value for toxin production was maintained. A previous study has reported that the amounts of agitation and aeration are important factors when cultivating fungus in the fermenter to produce chitinolytic enzyme. A mass production of diphtheria toxoid with a purity level greater than 2,500 Lf/ mgPN was obtained through purification and detoxification from this optimized toxin production.     

The immunization of adults against diphtheria in Sweden

Two hundred and three women who disclaimed vaccination against diphtheria were divided into four groups and injected with either 2.0 or 6.25 Lf of a routine diphtheria toxoid or of a more purified preparation. One hundred and twenty-six of these women who did not show a secondary antibody response were given a second and a third injection one month and one year, respectively, after the first injection. Prebooster (third injection) antitoxin titres of greater than or equal to 0.01 IU ml-1 (the minimum level for protection) were found in 22 and 37% of those who received 2.0 and 6.25 Lf, respectively. Postbooster titres of greater than or equal to 1.0 IU ml-1 (calculated to give a protection of at least ten years of duration) were found in 23 and 58% of those who received 2.0 and 6.25 Lf, respectively. The rate of untoward reactions was low. Fever of short duration occurred in five women. Four out of the five women received 6.25 Lf of the more purified diphtheria toxoid and one 2 Lf of the routine toxoid. Local reactions greater than 10 cm were observed in three women. All received the higher dose, 6.25 Lf of diphtheria toxoid. Local reactions greater than 5 but less than or equal to 10 cm occurred in up to 13% (6.25 Lf of diphtheria toxoid). No significant difference between the groups of women vaccinated with routine or more purified toxoid was found. It was concluded that the diphtheria toxoids in the two doses of 2 Lf and 6.25 Lf did not induce a satisfactory immune response. To induce adequate protection the dose of diphtheria vaccine needs to be the same for adults and children, i.e. 12.5 Lf.