百日咳杆菌的抗原

鲍特氏菌属的抗原共发现３大类：凝集原,丝状血凝素（ＦＨＡ）及毒素。在凝集原中只有７是百日咳,副百日咳及支气管败血症菌所共有,其他１３种各异。ＦＨＡ与百日咳毒素（ＰＴ）为无细胞苗的主要成分。毒素有５种,最主要的是ＰＴ,其他有不耐热毒素,气管细胞毒（ＴＣＴ）,脂多糖（ＬＰＳ）内毒素及腺嘌呤环化酶毒素（ＡＣＴ）。     

百日咳毒素单克隆抗体在抗感染中的应用

百日咳毒素（ＰＴ）单克隆抗体（ＰＴ－ＭｃＡｂ）具有特异性中和体外和体内百日咳毒素的促进白细胞增多（ＬＰ）、组胺敏感（ＨＳ）、胰岛激活（ＩＡ）、腺苷三磷酸核糖基转移酶（ＡＤＰ－Ｒ）、ＣＨＯ细胞簇聚等多种生物学活性的特性,同时亦具有防御百日咳杆菌脑内和气溶胶攻击的保护作用。     

一株减毒百日咳鲍特菌的构建及生物学特性分析

百日咳是传染性强、感染率高的急性呼吸道传染病,主要感染婴幼儿,是婴儿死亡的主要原因之一。百日咳鲍特菌（Bordetella pertussis）是引起百日咳的最主要病原菌。近年来世界各地多次出现百日咳暴发,迫切需研制更加有效的新型百日咳疫苗。本研究构建了一株减毒百日咳活疫苗BPTM1,利用同源重组方法敲除编码百日咳鲍特菌主要毒力因子百日咳毒素（pertussis toxin,PTX）和皮肤坏死毒素（dermonecrotic toxin,DNT）的基因,并用大肠埃希菌的同源基因置换了负责气管细胞毒素（tracheal cytotoxin,TCT）转运的基因ampG。通过聚合酶链反应验证了毒素及相关基因的敲除和置换,蛋白免疫印迹法检测表明PTX的S1亚基未表达。体外生长曲线和体内定植曲线均表明,相比于野生型百日咳鲍特菌BPMM,减毒BPTM1的生长和定植能力未受影响,其所致肺部病理效应减轻,而所诱导的百日咳鲍特菌特异性IgG、IgG1、IgG2a抗体保持高水平。本研究表明,减毒百日咳鲍特菌BPTM1有可能成为百日咳疫苗的候选疫苗。     

无细胞百日咳菌苗纯度和生物学特性研究

本文对无细胞百日咳菌苗的纯度、免疫力和毒性进行了研究。实验证明菌苗中不仅含有百日咳毒素（ＰＴ）和丝状血凝素（ＦＨＡ）,而且还含有百日咳凝集素和百日咳粘着素。菌苗的纯度为５０％－９５．７％,菌苗中ＰＴ和ＦＨＡ的比例为１：１－１：１８。     

吸附精制百日咳菌苗、白喉、破伤风类毒素混合制剂稳定性的研究

将连续生产并已申报人体接种反应观察的三批吸附精制百日咳菌苗、白喉、破伤风类毒素混合制剂,分别于４～８℃、１８～２２℃、３７℃贮存半年、１年、１年半和３０天,然后以小鼠法或豚鼠法对该制剂进行安全稳定性、效力稳定性以及毒性逆转等测试。其毒力安全稳定性试验表明,三批混合制剂中的精制百日咳菌苗在４～８℃贮半年、１年、１年半后、它们的ＢＷＤＵ／ｍｌ、ＬＰＵ／ｍｌ、ＨＳＵ／ｍｌ均无显著性差异（Ｐ＞０．０５）;白喉及破伤风类毒素在４～８℃贮存半年,１年、１年半以及１８～２２℃贮存半年和３７℃贮存３０天后,三批测试组的豚鼠均健存且体重增加。在效力稳定性试验中,４～８℃贮存１年和１年半后精制百日咳菌苗的效力与贮存前（分别为２６．７３ＩＵ／ｍｌ、２１．０２ＩＵ／ｍｌ、１９．１７ＩＵ／ｍｌ）无显著性差异（ｔ＝０．１１４;Ｐ＞０．０５）;白喉及破伤风类毒素４～８℃贮存长达１年半后,效力仍在２～３．２ＩＵ／ｍｌ,动物保护率仍可达８０～１００％。三批混合制剂在３７℃贮存３０天后,通过Ｓｃｈｉｃｋ毒性试验及测定ＢＷＤＵ／ｍｌ、ＬＰＵ／ｍｌ、ＨＳＵ／ｍｌ各指标显示,精制百日咳菌苗、白喉和破伤风类毒素无毒性逆转。说明该吸附精制百、白、破混合     

消灭传染病的国际协作(续四) 百日咳 白喉 破伤风联合疫苗

消灭传染病的国际协作（续四）　百日咳　白喉　破伤风联合疫苗木材三生夫（日本东海大学名誉教授）　春美（日本东海大学小儿科）为了让全世界儿童都能得到疫苗接种而开展的扩大预防接种计划（ＥＰＩ）已经取得了相当的成绩。下一个目标乃是为了尽可能地消灭那些可用疫苗...     

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...     

破伤风毒素C部分的克隆及在大肠杆菌中的表达

用ＣＲ方法从破伤风杆菌基因组ＤＮＡ上坟增出破伤风毒素Ｃ部分（ｔｅｔａｎｕｓ ｔｏｘｉｎ ｆｒａｇｍｅｎｔ Ｃ,ＴＴＣ）,经测序其基因片段由１３５６ｂｐ组成,可编码４５１个氨基酸残基的多肽。将其插入原核表达载体ｐＥＴ－２８ａ（＋）中,并在大肠杆力ＢＬ２１９ＤＥ３）中表达,其表达量占可溶性总蛋白质的８．２％;ＳＤＳ－ＰＡＧＥ和免疫印迹分析表明,ＴＴＣ基因（该基因ＧｅｎＢａｎｋ登录号为ＡＦ１５４８２）表     

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。     

破伤风毒素保护性抗原在毕氏酵母中的分泌表达

采用PCR方法．从C.Tetani 64008菌株中扩增大小为1353bp的破伤风毒素与靶细胞起结合作用的重链C端基因(Tetc),直接连接pGEM-T栽体进行测序,并以pPIC9K为表达栽体构建重组表达质粒,经线形化的重组质粒电转化毕氏酵母细胞GS1l5和KM71．甲醇诱导获得了分泌表达,表达产物存在于培养上清中,占分泌蛋白的10％,通过免疫印迹可以检测到重组表达产物,活性测定表明,重组蛋白具特异结合活性,本研究通过实现破伤风毒素保护性抗原在酵母系统中的分泌表达、研究其影响因素,为其它细菌毒素蛋白高效可溶性表达,及进一步抗原片段介导保护性免疫研究及抗毒素制备奠定了基础。     

流行性出血热免疫球蛋白的研制

本文首次报告采用纯化灭活流行性出血热（ＥＨＦ）Ⅰ型疫苗免疫健康献血员,采集ＥＨＦ抗体高滴度的血浆,用低温乙醇法及盐析法分离纯化三批ＥＨＦ免疫球蛋白。结果表明：（１）采用０,１,３,４（月）或０,１,４,５（月）免疫程序,疫苗剂量１～２ｍｌ（０．１５～０．３０ｍｇ蛋白）,受免献血员血清平均抗体滴度可达１∶４０６（ＥＬＩＳＡ）或１∶１１２（ＲＰＨＩ）。（２）通过生化检定,三批制品的电泳纯度为９７．０５％,９６．８４％,９９．２６％;ＩｇＧ单体和二聚体含量为８９．５５％,９１．３０％和９８．２１％。（３）用空斑抑制中和试验及免疫印染试验证明所纯化的免疫球蛋白具有抗ＥＨＦ病毒特异性。（４）三批ＥＨＦ免疫球蛋白的效价测定,结果为ＥＬＩＳＡ滴度≥１∶５１２,ＲＰＨＩ滴度≥１∶１０２４,ＰＲＮＴ（中和抗体）滴度１∶４０。按１６％蛋白计,ＥＨＦ免疫球蛋白的效价可达原料血浆的１０倍以上。（５）无菌、安全、毒性及热原质试验检定结果,全部通过《中国生物制品规程》要求。     

The relationship between airway immunoglobulin activity and eosinophils in COPD

In chronic obstructive pulmonary disease (COPD), the effects of inhaled corticosteroids are predicted by blood eosinophil counts. We previously briefly reported increased immunoglobulin (Ig)A and IgM levels in bronchoalveolar lavage (BAL) of COPD patients with higher (eosinophil^high) compared to lower (eosinophil^low) blood eosinophils (>250/μL versus < 150/μL), suggesting differences in adaptive immune function. An inverse relationship exists between eosinophil counts and airway pathogenic bacteria levels. The mechanistic reasons for these associations between eosinophils, corticosteroids and pathogenic bacteria are unclear. IgA, IgM and IgG levels were assessed in BAL, bronchial biopsies and epithelium collected from eosinophil^high (n = 20) and eosinophil^low (n = 21) patients. Bronchial B-cell numbers were measured by immunohistochemistry. B-cell activity was assessed in bronchial samples and following exposure to BAL from eosinophil^high and eosinophil^low patients. BAL levels of non-typeable Haemophilus influenza (NTHi)-specific immunoglobulins were quantified. Results showed airway expression of IgA, IgG1 and IgM were lower in eosinophil^low compared to eosinophil^high patients, with lower levels of NTHi-specific IgA and IgM. Bronchial B-cell numbers were similar in both groups, but B-cell activity was lower in eosinophil^low patients. In conclusion, COPD eosinophil^low patients show differences in adaptive immune function compared to COPD eosinophil^high patients. These differences may cause different microbiomes in these COPD phenotypes.     

Chicken liver ribosomes: Characterization of cross-reaction and inhibition of some functions by antibodies prepared against Escherichia coli ribosomal proteins L7 and L12

Antibodies prepared in rabbits against Escherichia coli ribosomal proteins L7/L12 are reported to be immunologically cross-reactive with some ribosomal proteins on the 60 S subunit of eukaryote ribosomes (Wool & Stöffler, 1974; Stöffler et al., 1974). We have confirmed these reports and extended this finding to a detailed study of the functional properties of eukaryote ribosomes which are affected by these cross-reacting antibodies. We report here the partial reactions in protein synthesis that are inhibited by the anti-L7/L12 IgG (immunoglobulin G) preparations using a chicken liver system. The following reactions were inhibited: EF-1 (elongation factor 1) dependent binding of aminoacyl-tRNA to ribosomes and GTP hydrolysis; EF-2 dependent binding of nucleotide to ribosomes and GTP hydrolysis; binding of [¹⁴C]ADP-ribosyl · EF-2 to ribosomes. This last reaction is more sensitive to the antibody inhibition than the corresponding nucleotide binding reaction. We show that the inhibitions were not simply non-specific precipitation of ribosomes by IgG, in that monovalent Fabs were also inhibitory, and peptidyl transferase activity was not inhibited. The functions inhibited with the IgG preparations in the chicken liver system are analogous to those inhibited in the homologous E. coli system. Thus the cross-reacting protein is functionally as well as immunologically conserved.     

Pyriproxyfen enhances the immunoglobulin G immune response in mice

Pyriproxyfen is a juvenile hormone mimic of vital importance for insect development with little risk to humans. This study was performed to investigate whether large doses of pyriproxyfen affect the immune response in mammals. Mice were immunized thrice with ovalbumin in 5% ethanol, with or without pyriproxyfen or alum. Large doses of pyriproxyfen (9 or 15 mM) significantly enhanced specific total IgG immune response. This enhancement was no longer present 24 hr after treatment with pyriproxyfen. These results suggest that pyriproxyfen is a safe chemical. Moreover, pyriproxyfen induced higher titers of IgG2a and enhanced tumor necrosis factor‐alpha and gamma‐interferon responses whereas alum induced IgG1 with enhanced interleukin‐4 and ‐10. These observations indicate that the mechanism of immune enhancement by pyriproxyfen may differ from that of alum.     

A comparative study of the effect of modification of the surface of human platelets on the receptors for aggregated immunoglobulins and for ristocetin-von Willebrand factor

The receptors for aggregated immunoglobulin G (IgG) (an Fc receptor) and for ristocetin-von Willebrand factor on human platelets were studied by means of various modifications of the platelet surface. The expression of these receptors was measured by the agglutination of platelets to ristocetin in the presence of von Willebrand factor, which is part of the factor VIII complex, and by the binding of aggregated IgG coupled to ³H-labelled diazobenzen. Treatment of platelets with chymotrypsin, trypsin, papain and pronase which removed protein and glycoprotein from the platelet under conditions where the release reaction was inhibited caused loss of the expression of the receptor for ristocetin-von Willebrand factor and an enhancement of that for aggregated IgG. Induction of membrane changes with ADP and of the release reaction with the ionophore A23187 abolished agglutination to ristocetin-von Willebrand factor but did not alter the receptor for aggregated IgG. Possible contributions of unspecific membrane changes, produced by protease treatment of platelets, to the modification of receptor expression were eliminated by the use of formaldehyde-treated platelets. Trypsin, papain and pronase destroyed the ability of these platelets to agglutinate to ristocetin-von Willebrand factor but produced no change in the binding of aggregated IgG. Therefore, the receptor for ristocetin-von Willebrand factor is truly sensitive to proteolysis while the Fc receptor is not, but is partially masked by protease-sensitive material.     

分泌型免疫球蛋白A的研究进展

大部分感染都起源于黏膜表面,而黏膜免疫的主要抗体是分泌型免疫球蛋白A（SIgA）,它能有效地阻断病原体的感染和侵入。SIgA是由1个IgA二聚体、1条J链和1个分泌片（SC）共价结合构成的异源十聚体。IgA和J链由活化B细胞产生,SC则由黏膜上皮细胞合成。SIgA分子具有极高的稳定性和极强的抗微生物活性。我们就SIgA合成的相关机制、IgA单体和SIgA的结构与功能,以及重组SIgA的研究进展简要综述。     

Immunoglobulin concentration in Atlantic cod, Gadus morhua L., serum and cross-reactivity between anti-cod-antibodies and immunoglobulins from other species

The immunoglobulin (Ig) level in serum from Atlantic cod, Gudus morhua L. (Teleostomi, Gadiformes), was measured using ELISA. Cod serum contains 5.62 ± 0.19 mg Ig ml, which constitute 17.2% of the total serum proteins.
Polyclonal and monoclonal antibodies (Ab) against cod-Ig were used for cross-reactivity studies in sera from other species. The cross-reaction with polyclonal Ab was high in gadifom fish, and low or absent in the other tested species. Immunoprecipitation showed that the Ab only binds to the Ig of the serum from the other species. The results within the gadiform group indicate that (1) Gadidae and Lotidae are more closely related to each other than to Merluccidae, (2) Phycidae is more distant related to Lotidae than earlier considered, (3) Zoarcidae seem to be more distant related to Gadus than other species of Gadiformes. These results support recent revisions of the systematics of Gadiformes.
Monoclonal Ab against the heavy chain of cod-Ig show no, or small (< 15%) cross-reactions with serum from gadiform species. One monoclonal Ab against the light chain shows high cross-reaction and also binds to Ig from species outside the Gadiformes, indicating that the light chain of Ig might be more conserved than the heavy chain.     

Characterisation of a chromatographically produced anti-D immunoglobulin product

A chromatographic fractionation method has been developed for the production of a liquid-stable anti-D immunoglobulin product for intravenous and intramuscular use. An immunoglobulin fraction, highly enriched with anti-D immunoglobulins, was isolated by cation-exchange column chromatography and further polished, first by anion-exchange chromatography, followed by an aluminium hydroxide gel treatment. The process includes two specific steps for virus inactivation and removal, namely S/D treatment and nanofiltration. The overall anti-D process yield is about 56%. The final product is stabilised with human albumin and glycine and placed in ready-to-use syringes. The anti-D product was shown to be stable in liquid state for at least 30 months at 4°C.     

冻干静脉注射人血丙种球蛋白的研制

作者参照瑞典ＫａｂｉＶｉｔｒｕｂ药厂的“Ｇａｍｍｏｎａｔｉｖ”的制备方法,以低温乙醇法的ＦⅡ沉淀为原料,经ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ－５０吸附,加入人血白蛋白作为稳定剂制备静脉注射人血丙种球蛋白（ＩＶＩＧ）。该制剂的γ－球蛋白纯度＞９４％,ＩｇＧ单体二聚体含量＞９７％,ＩｇＧ各亚类均存在且构成比与正常人血浆相似。抗补体活性（ＡＣＡ）和前激肽释放酶激活剂活性（ＰＫＡ）符合规程要求,安全性和稳定性好。该制剂为冻干剂型,便于运输和保存。     

Recent findings on the structure and function of teleost IgT

As key effector molecules of jawed vertebrate’s adaptive immune system, immunoglobulins are produced by B lymphocytes, either as a secretory form (antibody) or as a membrane form (B cell receptor). Until recently, teleost fish B cells were thought to express only two classes of immunoglobulins, IgM and IgD. In addition, IgM in these species was thought to be the only immunoglobulin isotype responding to pathogens both in systemic or mucosal compartments. However, the unexpected discovery of IgT, a new teleost immunoglobulin unearthed in 2005, has provided for new opportunities to analyze further roles of teleost immunoglobulins in these two physiologically distinct compartments. The smoke about the potential function of IgT has cleared recently with the finding that this immunoglobulin appears to be specialized in gut mucosal immunity. Significantly, the new capability of measuring not only IgM but also IgT responses will greatly facilitate the evaluation and understanding of fish immune responses as well as the protective effects of fish vaccines. The purpose of this review is to summarize the molecular characterization of new IgT orthologs and subtypes in teleosts, as well as to describe the new findings concerning the protein structure of IgT, the B cells producing it, and its role in mucosal immunity.