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。     

不同纯度、剂量白喉类毒素对迟发型超敏反应的影响

研究不同剂量及不同白喉类毒素纯度引起的迟发型超敏反应的状况,用于指导疫苗的生产,提高疫苗的质量。以豚鼠为动物模型,采用迟发型超敏试验法,对原制白喉类毒素、精制白喉类毒素、纯化精制白喉类毒素、高纯度的精制白喉类毒素的剂量与超敏反应试验。试验结果表明,注射白喉类毒素剂量的大小与超敏反应成正相关,与纯度成负相关。剂量越大,超敏反应越强;纯度越高,超敏反应愈弱。     

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…     

CpG对乙型肝炎基因重组(CHO细胞)疫苗免疫效果的影响

为了研究CpG-寡脱氧核苷酸(CpG-OPN)作为佐剂对乙型肝炎基因重组(CHO细胞)疫苗(简称乙肝疫苗)免疫效果的影响,以乙肝疫苗加Al(OH)3、疫苗加CpG和疫苗加Al(OH)3与CpG3三种配伍方式,通过腹腔、皮下或肌内3种不同途径免疫Balb/c小鼠,观察不同免疫途径和不同配伍的免疫效果.同时又将疫苗与CpG混合后在4℃存放6个月再免疫小鼠,观察CpG的稳定性.结果表明:①3种免疫途径中以肌内注射效果最好,这在使用CpG的实验组尤为明显,在该组肌内免疫的ED50比腹腔的低了10倍,而诱发的抗体滴度提高了3倍;②疫苗与CpG、Al(OH)3联合使用的免疫效果最好,在肌内免疫时联合使用的免疫效果比疫苗+Al(OH)3提高4倍,比疫苗+CpG提高7倍;③疫苗+Al(OH)3免疫时,表现为IgG1抗体亚型占优势,而再加入CpG后则IgG1和IgG2a均升高,以IgG2a最显著;④疫苗与CpG混合后4℃保存半年,不影响其活性.     

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     

钙调磷酸酶B亚单位(CnB)作为佐剂对新型乙型肝炎病毒疫苗免疫效果的影响

比较不同佐剂配伍的效果,探讨常规剂量(5μg)钙调磷酸酶B亚单位(Calcineurin subunit B,CnB)佐剂对含PreS1+S融合抗原的乙型肝炎病毒新型疫苗(HBSS1)免疫效果的影响。采用Al(OH)3、常规剂量(5μg)CnB及CnB+Al(OH)3等佐剂与HBV颗粒疫苗配伍初次免疫,重组腺病毒载体疫苗加强免疫的策略,在C57BL/6小鼠模型上研究不同佐剂对HBV颗粒疫苗肌肉注射后免疫应答的影响,主要包括抗体滴度、抗体亚型分类及特异性细胞免疫(γ-IFN ELISpot检测)。研究结果显示Al(OH)3佐剂存在明显免疫增强作用,而单独加入5μg CnB佐剂或CnB与Al(OH)3佐剂联合应用对Anti-PreS1抗体无明显的增强作用,但可显著降低anti-HBs抗体水平;各免疫组在重组腺病毒载体疫苗加强后,其抗体亚类包括IgG1、IgG2a和IgG2b;并可诱导高水平的细胞免疫应答反应。因而常规剂量(5μg)CnB单独或联合Al(OH)3佐剂对新型HBV疫苗无明显的免疫增强作用。     

钙调磷酸酶B亚单位(CnB)作为佐剂对新型乙型肝炎病毒疫苗免疫效果的影响北大核心CSCD

比较不同佐剂配伍的效果,探讨常规剂量(5μg)钙调磷酸酶B亚单位(Calcineurin subunit B,CnB)佐剂对含PreS1+S融合抗原的乙型肝炎病毒新型疫苗(HBSS1)免疫效果的影响。采用Al(OH)3、常规剂量(5μg)CnB及CnB+Al(OH)3等佐剂与HBV颗粒疫苗配伍初次免疫,重组腺病毒载体疫苗加强免疫的策略,在C57BL/6小鼠模型上研究不同佐剂对HBV颗粒疫苗肌肉注射后免疫应答的影响,主要包括抗体滴度、抗体亚型分类及特异性细胞免疫(γ-IFN ELISpot检测)。研究结果显示Al(OH)3佐剂存在明显免疫增强作用,而单独加入5μg CnB佐剂或CnB与Al(OH)3佐剂联合应用对Anti-PreS1抗体无明显的增强作用,但可显著降低anti-HBs抗体水平;各免疫组在重组腺病毒载体疫苗加强后,其抗体亚类包括IgG1、IgG2a和IgG2b;并可诱导高水平的细胞免疫应答反应。因而常规剂量(5μg)CnB单独或联合Al(OH)3佐剂对新型HBV疫苗无明显的免疫增强作用。     

HFRS疫苗佐剂吸附前后加入保护剂对其免疫原性的影响

为提高肾综合征出血热(HFRS)双价纯化疫苗抗原的有效利用率,在保证疫苗免疫效果的前提下,合理地降低疫苗的成本,确定最佳疫苗配制方法和最合适的疫苗浓缩倍数。将抗原含量不同的出血热纯化疫苗原液及疫苗配制的两种主要成分———人血白蛋白和A l(OH)3佐剂按不同的配制顺序配制成4批纯化疫苗,分别抽样,测其上清液中抗原量并做效力试验。结果表明人血白蛋白和A l(OH)3佐剂加入的顺序不同,配制的4批纯化疫苗上清液抗原量也不同,效力试验结果显示其中和抗体阳转率(PRNT)也有很大差别。因此在纯化疫苗的配制过程中,人血白蛋白和A l(OH)3的加入顺序影响到佐剂A l(OH)3对疫苗抗原的吸附,也影响到疫苗的免疫效果;先加A l(OH)3再加人血白蛋白的疫苗免疫效果要好于先加人血白蛋白再加A l(OH)3的疫苗。     

儿童初次免疫后25—30年抗白喉的免疫力

<正>丹麦对儿童进行白喉初次免疫,接种含有白喉类毒素(50lp/ml)和破伤风类毒素(12 1/2Lf/ml)的氢氧化铝吸附制剂1毫升,自1950年以来作为常规使用。1970年,白、破制剂中又加入了脊髓灰质炎疫苗。推荐儿童初次免疫必须分三针注射:第5个月、第6个月和第15个月。丹麦一般不进行加强免疫,唯在军人中推行常规免疫和加强     

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。     

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.     

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.     

Comparative study of the immunoglobulins and specific antibodies in the blood of rabbits immunized with ADTP vaccine and ADT-anatoxin with normal and reduced antigen contents

Adsorbed DPT vaccine and adsorbed DT toxoids with normal and reduced antigen content were used for the immunization of rabbits. The levels of IgM and IgG and the dynamics of antibodies to diphtheria and tetanus toxins and to Bordetella pertussis in the blood sera of the animals were studied in the postvaccinal period (on days 15 and 34). This study revealed that the reduction of the antigen content of adsorbed DT toxoid to 5 Lf of diphtheria toxoid and 5 binding units of tetanus toxoid did not decrease the capacity of the preparation for increasing the levels of IgG and IgM, antibodies to diphtheria and tetanus toxins in the sera of the rabbits. The reduced content of these toxoids in adsorbed DPT vaccine did not affect its capacity for inducing the enhanced synthesis of IgG, antibodies to diphtheria and tetanus toxins, while the production of IgM and IgA remained unchanged. At the same time an increase in the titers of antibodies to B. pertussis in the animals was less pronounced than that observed after the injection of commercial adsorbed DPT vaccine. Additional investigations are necessary in order to establish the protective potency of the pertussis component in adsorbed DPT vaccine with the reduced content of toxoids and to find out the optimum antigenic composition for this preparation.     

The partially degraded lipopolysaccharide of Burkholderia cepacia and ornithine-containing lipids derived from some Gram-negative bacteria are useful complex lipid adjuvants

The partially degraded lipopolysaccharide of Burkholderia cepacia (LPSdegr) and the ornithine-containing lipids were purified from some bacteria. The substances were developed as complex lipid adjuvants, because they have weak toxicity and are able to activate the immune systems of the living body. After various toxoid antigens such as pertussis toxoid, diphtheria toxoid and tetanus toxoid were mixed with the complex lipid adjuvants, the mixtures were administered to mice subcutaneously. Antitoxoid IgG antibody titers in the serum were measured several times over 3 months. The efficacy of the LPSdegr as adjuvant was almost as high as that of the ornithine-containing lipids, and it was almost equal to that of the aluminum hydroxide adjuvant (Alum), which is generally used as a vaccine adjuvant.     

Immune response in adult immunized with adsorbed DT-M anatoxin with reduced antigen content and Imovax-DT-adulte vaccine

The comparative study of immune response after immunization of adults with adsorbed DT toxoid with reduced antigen content and Imovax-DT-adulte vaccine, as well as the safety of these preparations, was made. The study revealed that immunization of adults with adsorbed DT toxoid having reduced antigen content, made in two injections, and the injection of Imovax-DT-dulte vaccine, as well as the successive injection of these preparations, produced the same the levels of antitetanus immunity. Antidiphtheria immunity, evaluated by the number of seroconverted to diphtheria persons following two injections immunization was similar for the two preparations, while the level of antidiphtheria antibodies was higher in persons immunized with adsorbed DT toxoid. The immune stratum index was rather high among persons aged 16-29 years. This age group exhibited the highest number of persons, seropositive to both diphtheria and tetanus. Both vaccine preparations, adsorbed DT toxoid with reduced antigen content and Imovax-DT-adulte vaccine, were found to be equally capable of inducing autoimmune reactions in the vaccinees, detected by laboratory methods.     

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。     

The ecological aspects of ADTP vaccination]

The results of measuring the activity of diphtheria and tetanus toxoids in the blood of 19,654 persons immunized with adsorbed DPT vaccine have shown a pronounced decrease in the effectiveness of booster immunization with adsorbed DPT vaccine. The negative influence of environmental pollution is more pronounced with respect to less active diphtheria toxoid. The possibility of the separation of the population into strata differing in the intensity of their immune response to toxoids under the influence of environmental pollution is considered.     

Liposomal Hepatitis A Vaccine and Liposomal Multiantigen Combination Vaccines

Abstract

Many antigens are only weak immunogens, and they require adjuvants to reach a satisfactory immune response as vaccines. This is especially true for modern antigens, such as products of gene technology or synthetic peptides. Also, classically produced antigens such as diphtheria or tetanus-antigens are coupled on aluminium phosphate or aluminium hydroxide – to this day practically the only adjuvant class which is routinely used in vaccinology. Aluminium phosphate was first used in the 1920s as an immunopotentiator for the diphtheria vaccine. Since then, and up until now, it has not been possible to fully replace this unsatisfactory adjuvant. exclusion of epitope-specific suppression by the individual components, it was evident that particularly the diphtheria and tetanus constituents exerted “antigenic competition” on the HAV antigen and possibly also on the HBs antigen. By reducing the diphtheria and tetanus toxoid subunit molecules per IRIV particle, it was possible not only to increase significantly the immune response to these antigens but also to remove completely the antigenic competition on the HAV and HBs antigens. Once the optimum composition of the vaccine had been achieved (careful dosing of antigens per IRIV particle), an immunological effect clearly superior to that of comparable, aluminium-adsorbed products was obtained (Table II). The tolerability of this “supercombi vaccine” was significantly better than the commercial alum adsorbed products (Table III).     

Determination of the antigenic activity of diphtheria anatoxin in liquid and sorbed preparations using an erythrocyte diphtheria antigen (anatoxin) diagnostic agent

On the basis of the experimentally established dependence of the degree of binding of diphtheria toxoid with standard diphtheria antitoxin on the duration of their joint incubation with the maximum binding occurring in 3-4 hours of incubation at 37 degrees C, the method of the in vitro determination of the antigenic activity of diphtheria toxoid in liquid and adsorbed preparations is proposed. The method is based on the principle of double binding with the use of diphtheria antigenic (toxoid) erythrocyte diagnosticum. The antigenic activity of diphtheria toxoid, evaluated by the degree of its maximum binding with diphtheria antitoxin, correlated with its antitoxin-binding activity in animal experiments and did not correlate with its flocculating activity. The antigenic activity of diphtheria toxoid in adsorbed preparations, evaluated by their maximum binding with standard diphtheria antitoxin, was shown to be closely related to the immunogenic potency of these vaccines in animals.