Antibody response to dengue virus

In this review, we discuss the current knowledge of the role of the antibody response against dengue virus and highlight novel insights into targets recognized by the human antibody response. We also discuss how the balance of pathological and protective antibody responses in the host critically influences clinical aspects of the disease.     

抗脱氧雪腐镰刀菌烯醇单克隆抗体的制备

[目的]小麦赤霉病菌产生的毒素不仅在病害发展过程中具有加重赤霉病的作用,而且污染谷物导致严重的食用安全性问题.由于赤霉病的普遍发生,有必要建立快速、灵敏、有效的毒素检测方法,本试验旨在制备可用于检测被脱氧雪腐镰刀菌烯醇污染的粮谷类特异性单克隆抗体.[方法]本实验首先将脱氧雪腐镰刀菌烯醇(DON)的衍生物3-半琥珀酰-脱氧雪腐镰刀菌烯醇(3-HS-DON-OVA)与卵清蛋白(OVA)采用碳化二亚胺法进行偶联得到人工抗原,以此人工抗原免疫BALB/C小鼠,取该鼠脾细胞与SP2/O鼠骨髓瘤细胞融合,经筛选和克隆,得到了1株能稳定分泌DON抗体的单克隆细胞株(382),并制备单克隆抗体腹水.[结果]经检测382的抗体类型及亚类均为IgG1,其轻链为κ链.腹水通过间接酶联免疫吸附测定效价在1×10-7以上.该单克隆抗体与脱氧雪腐镰刀菌烯醇特异性结合反应的50%抑制质量浓度为29 μg/L,除与3-acetyldeoxynivalenol(3-Ac-DON)的交叉反应率为78.38%,与其他脱氧雪腐镰刀菌烯醇结构类似物无交叉反应.[结论]本实验所制备的单克隆抗体有较高的灵敏度和特异性,具有较好的应用价值.     

经蛋白A连接于琼脂糖的单克隆抗体亲和层析柱用于抗独特型抗体的纯化

 以Sepharose CL-4B-Pro A吸附胃癌单克隆抗体(McAb)PD4,继之以交联剂二甲基庚二亚胺二盐酸盐(dimethyl pimelimidate dihydrochloride)处理,形成亲和介质(柱Ⅰ)。该亲和介质用于纯化抗PD4独特型抗体(aIdAb)具有明显的优点。与Sepharose CL-4B-PD4(柱Ⅱ)相比,前者的结合容量为后者的1.78～1.94倍。采用柱Ⅰ纯化的aIdAb,当其与McAb PD4的分子比分别为1:1及4:1时可50％或100％地抑制McAb PD4与靶细胞的结合,但采用柱Ⅱ获得的aIdAb,只有当分子比达到2:1及8:1时,才能达到同等的抑制效应。这可能是由于Pro A与McAb PD4的Fc片断结合,使后者的Fab端得以充分暴露,因而有更多的机会与aIdAb结合。     

Antibody engineering to develop new antirheumatic therapies

There has been a therapeutic revolution in rheumatology over the past 15 years, characterised by a move away from oral immuno-suppressive drugs toward parenteral targeted biological therapies. The potency and relative safety of the newer agents has facilitated a more aggressive approach to treatment, with many more patients achieving disease remission. There is even a prevailing sense that disease 'cure' may be a realistic goal in the future. These developments were underpinned by an earlier revolution in molecular biology and protein engineering as well as key advances in our understanding of rheumatoid arthritis pathogenesis. This review will focus on antibody engineering as the key driver behind our current and developing range of antirheumatic treatments.     

A Monoclonal Antibody to Rabbit Brain GABA Transaminase

A monoclonal antibody of class IgG (subclass IgG1) has been prepared to rabbit brain GABA transaminase (GABA-T). This antibody reveals a single band of molecular weight 52,000 on a nitrocellulose filter blotted with purified GABA-T. On a filter blotted with unfractionated rabbit brain supernatant a major band of molecular weight 58,000 is revealed. An immunoaffinity column was prepared by coupling proteins from ascites fluid containing anti-rabbit GABA-T antibody to Bio-Rad Affi-Gel 15. This column bound purified GABA-T and extracted from unfractionated rabbit brain supernatant a protein of molecular weight 58,000, which was almost homogeneous and which had GABA-T enzyme activity. Using immunoaffinity chromatography, therefore, a high degree of purification of GABA-T may be achieved in a single step. Further, this technique may preserve an authentic form of the enzyme that is lost during the conventional purification procedure. The antibody inhibits GABA-T enzyme activity, up to a maximum of 35%.     

Prevalence of Antibody to Current Influenza Viruses and Effect of Vaccination on Antibody Response

The extent of antibody to the influenza virus A/Hong Kong/68 (H3N2) after four years of prevalence was investigated in Britain and in the U.S.A. The results indicated a high incidence in both populations. The prevalence of antibody to a variant A/England/42/72 (H3N2) which has been causing epidemics of influenza in the southern hemisphere during the middle months of 1972 was also investigated. The differences reflect the shift in antigenic content of this variant, and although the overall proportion of the sera with antibody at > 1/40 was 37%, some age groups had an incidence of only 20% or less with antibody at this level. A commercial inactivated A/Hong Kong/68 influenza vaccine was given to a group of volunteers in Britain to see how effective it might be in stimulating antibody to the variant A/England/42/72. The antibody responses were better than expected from earlier vaccine studies, and 63% of the vaccinees developed antibody to the A/England/42/72 to levels thought likely to be protective. This suggested that until a vaccine made with the variant A/England/42/72 becomes available the present A/Hong Kong/68 vaccine would be of use to protect those at special risk this winter.     

Antibody response to Candida albicans cell wall antigens

The cell wall of Candida albicans is not only the structure where many essential biological functions reside but is also a significant source of candidal antigens. The major cell wall components that elicit a response from the host immune system are proteins and glycoproteins, the latter being predominantly mannoproteins. Both carbohydrate and protein moieties are able to trigger immune responses. Proteins and glycoproteins exposed at the most external layers of the wall structure are involved in several types of interactions of fungal cells with the exocellular environment. Thus, coating of fungal cells with host antibodies has the potential to profoundly influence the host-parasite interaction by affecting antibody-mediated functions such as opsonin-enhanced phagocytosis and blocking the binding activity of fungal adhesins to host ligands. In this review we examine various members of the protein and glycoprotein fraction of the C. albicans cell wall that elicit an antibody response in vivo. Some of the studies demonstrate that certain cell wall antigens and anti-cell wall antibodies may be the basis for developing specific and sensitive serologic tests for the diagnosis of candidiasis, particularly the disseminated form. In addition, recent studies have focused on the potential of antibodies against the cell wall protein determinants in protecting the host against infection. Hence, a better understanding of the humoral response triggered by the cell wall antigens of C. albicans may provide the basis for the development of (i) effective procedures for the serodiagnosis of disseminated candidiasis, and (ii) novel prophylactic (vaccination) and therapeutic strategies to control this type of infections.     

Validating Antibodies to the Cannabinoid CB2 Receptor: Antibody Sensitivity Is Not Evidence of Antibody Specificity

Antibody-based methods for the detection and quantification of membrane integral proteins, in particular, the G protein-coupled receptors (GPCRs), have been plagued with issues of primary antibody specificity. In this report, we investigate one of the most commonly utilized commercial antibodies for the cannabinoid CB2 receptor, a GPCR, using immunoblotting in combination with mass spectrometry. In this way, we were able to develop powerful negative and novel positive controls. By doing this, we are able to demonstrate that it is possible for an antibody to be sensitive for a protein of interest—in this case CB2—but still cross-react with other proteins and therefore lack specificity. Specifically, we were able to use western blotting combined with mass spectrometry to unequivocally identify CB2 protein in over-expressing cell lines. This shows that a common practice of validating antibodies with positive controls only is insufficient to ensure antibody reliability. In addition, our work is the first to develop a label-free method of protein detection using mass spectrometry that, with further refinement, could provide unequivocal identification of CB2 receptor protein in native tissues.     

Antibody response to sulfolipids in experimental tuberculosis

Live Mycobacterium tuberculosis H₃₇Rv, when injected into guinea pigs, induced antibodies to sulfolipids whereas antibodies were not detected in animals injected with heat killed cells. The antibody titre was found to be related to the degree of infection. A significant decrease in the titre was noted after streptomycin treatment, suggesting that antibodies were produced in response to an increasing bacterial load that occurred as the infection progressed.     

Antibody engineering

The antibody molecule is a therapeutic agent, designed by nature to bind to a wide range of antigen molecules and to trigger effector functions, such as complement lysis and cell-mediated killing. The genes encoding antibodies can be manipulated in vitro, allowing the binding sites for antigen and effector molecules to be dissected, and new properties to be engineered. The future for the application of engineered antibodies in medicine is reviewed in the context of the past century.     

Contribution of Peptide Backbone to Anti-Citrullinated Peptide Antibody Reactivity

Rheumatoid arthritis (RA) is one of the most common autoimmune diseases, affecting approximately 1–2% of the world population. One of the characteristic features of RA is the presence of autoantibodies. Especially the highly specific anti-citrullinated peptide antibodies (ACPAs), which have been found in up to 70% of RA patients’ sera, have received much attention. Several citrullinated proteins are associated with RA, suggesting that ACPAs may react with different sequence patterns, separating them from traditional antibodies, whose reactivity usually is specific towards a single target. As ACPAs have been suggested to be involved in the development of RA, knowledge about these antibodies may be crucial. In this study, we examined the influence of peptide backbone for ACPA reactivity in immunoassays. The antibodies were found to be reactive with a central Cit-Gly motif being essential for ACPA reactivity and to be cross-reactive between the selected citrullinated peptides. The remaining amino acids within the citrullinated peptides were found to be of less importance for antibody reactivity. Moreover, these findings indicated that the Cit-Gly motif in combination with peptide backbone is essential for antibody reactivity. Based on these findings it was speculated that any amino acid sequence, which brings the peptide into a properly folded structure for antibody recognition is sufficient for antibody reactivity. These findings are in accordance with the current hypothesis that structural homology rather than sequence homology are favored between citrullinated epitopes. These findings are important in relation to clarifying the etiology of RA and to determine the nature of ACPAs, e.g. why some Cit-Gly-containing sequences are not targeted by ACPAs.