杭州浙大迪迅生物基因工程有限公司

　　原标题：重组低致敏性B细胞表位型草花粉过敏疫苗BM32与过敏原提取物型疫苗的免疫原性比较
　　过敏原特异性免疫治疗(AIT)是临床治疗过敏的一种经济有效的方法，可改变疾病的进程，并具有长期的效果。然而，基于过敏原提取物形式的AIT需要多次给药，这使得治疗很麻烦，导致患者依从性差。为了解决这一问题，人们提出了许多方法，包括使用安全性更高的AIT材料，如类变应原、重组变应原衍生物和变应原衍生肽，这些材料可以缩短累积阶段
　　在本研究中，我们研究了重组B细胞表位过敏疫苗BM32诱导过敏原特异性IgG抗体的能力，以及这些抗体抑制过敏患者IgE与草花粉过敏原结合的能力以及过敏原诱导的T细胞增殖能力。BM32重组草花粉(Phleum pratense)过敏疫苗的组成基于4个融合蛋白肽，由4种梯牧草草花粉主要过敏原(Phl p 1, Phl p 2, Phl p 5和Phl p 6) 融合到来自乙型肝炎病毒的载体蛋白，因该载体蛋白无过敏活性,因此可被注入过敏患者体内而不必有剂量递增阶段。为研究免疫原性,我们用一个剂量(即4 个BM融合蛋白每个20μg) 对过敏病人进行安全注射 (临床试验批准号：NCT01538979 NCT01445002和NCT02643641)。
　　本研究的主要发现是：在兔子中，皮下注射氢氧化铝吸附的BM32三个月，诱导针对主要草花粉过敏原Phl p1, p Phl p5和Phl p 6 的IgG抗体水平相当于需要注射超过8次的已注册的基于天然过敏原提取物的草花粉过敏疫苗(Allergovit草;Allergopharma、Reinbek、德国;Alutard SQ混合草;ALK-Abello、Hørsholm、丹麦;以及光草+黑麦;Stallergenes、安东尼、法国;参见本文在线存储库中的方法部分，网址为www.jacionline.org;图1)所诱导的IgG抗体的水平，而Pollinex (Pollinex四极体+禾本科+黑麦，Bencard Allergie GmbH德国慕尼黑,一种基于4种注射剂的疫苗)几乎没有反应。重要的是，BM32诱导的phlp2特异性IgG抗体水平高于任何已注册的基于过敏原提取物的疫苗(图1)。
　　我们认为这是一个重要的发现,因为已证明第二组草花粉主要过敏原被60%以上草花粉过敏患者识别, 通过皮肤试验发现，比其他草花粉过敏原具有更高的过敏效力。因此，我们的研究结果表明，只需少量注射(即3-5次)BM32，就有可能建立足够水平的草花粉过敏原特异性IgG反应，而传统的过敏疫苗需要两倍以上的增加剂量注射。舌下治疗甚至需要每日服用。因此，我们认为基于BM32的治疗方案将更方便患者，并增加患者的依从性。
　　在IgE抑制实验中，BM32诱导的IgG阻断IgE与phlp2结合的效果明显好于商业疫苗诱导的IgG抗体(p = .0008，图2)，因此可以认为BM32在保护第二组过敏原引起的症状方面可能优于基于提取物的疫苗。最后，我们还可以证明BM32诱导的抗体可以抑制特定的草花粉过敏原诱导的T细胞增殖(参见本文的方法部分和图E3，见www.jacionline.org的在线知识库)，这表明BM32可能对T细胞介导的草花粉过敏的迟发相症状也有有益的影响。然而，我们的研究的一个局限性是，我们在原始动物中进行了免疫原性研究，因此可能对已经致敏的患者存在差异。
　　综上所述，我们的免疫原性研究表明，与天然的基于过敏原提取物的疫苗相比，诱导草花粉过敏原特异性保护性抗体反应所需的BM32注射量要少得多，而且在疫苗中使用载体蛋白有助于克服第二组过敏原免疫原性差的问题。鉴于BM32具有较强的抗过敏活性，人们可能认为BM32是一种优于基于过敏原提取物疫苗的草花粉过敏疫苗。
延伸阅读
JACI                                                                    
Comparison of the immunogenicity of BM32, a recombinant hypoallergenic B cell epitope–based grass pollen allergy vaccine with allergen extract–based vaccines
DOI: https://doi.org/10.1016/j.jaci.2017.03.048|
Abstract:
Allergen-specific immunotherapy (AIT) is a clinically and cost-effective allergy treatment that modifies the course of the disease and has long-lasting effects.1x1Cox, L., Calderón, M., and Pfaar, O. Subcutaneous allergen immunotherapy for allergic disease: examining efficacy, safety and cost-effectiveness of current and novel formulations. Immunotherapy. 2012; 4: 601–616
Crossref | PubMed | Scopus (31) | Google ScholarSee all References, E7xE7Jutel, M., Agache, I., Bonini, S., Burks, A.W., Calderon, M., Canonica, W. et al. International consensus on allergy immunotherapy. J Allergy Clin Immunol. 2015; 136: 556–568
Abstract | Full Text | Full Text PDF | PubMed | Scopus (201) | Google ScholarSee all References, E8xE8Jacobsen, L., Niggemann, B., Dreborg, S., Ferdousi, H.A., Halken, S., Host, A. et al. Specific immunotherapy has long-term preventive effect of seasonal and perennial asthma: 10-year follow-up on the PAT study. Allergy. 2007; 62: 943–948
Crossref | PubMed | Scopus (617) | Google ScholarSee all References However, allergen extract–based forms of AIT require administration of multiple doses, which makes treatment cumbersome and leads to poor compliance in patients.2x2Keil, M.A., Röder, E., Gerth van Wijk, R., Al, M.J., Hop, W.C., and Rutten-van Mölken, M.P. Real-life compliance and persistence among users of subcutaneous and sublingual allergen immunotherapy. J Allergy Clin Immunol. 2013; 132: 353–360
Abstract | Full Text | Full Text PDF | PubMed | Scopus (115) | Google ScholarSee all References A number of approaches were proposed to address this issue, including use of AIT materials with higher safety, such as allergoids, recombinant allergen derivatives, and allergen-derived peptides, which allow shortening of the build-up phase.E9 In this study we have investigated the ability of the recombinant B cell epitope–based allergy vaccine BM32 to induce allergen-specific IgG antibodies and the ability of these antibodies to inhibit allergic patients' IgE binding to grass pollen allergens, as well as allergen-induced T-cell proliferation. BM32 is a recombinant grass pollen (Phleum pratense) allergy vaccine based on 4 fusion proteins consisting of peptides from the 4 major timothy grass pollen allergens (Phl p 1, Phl p 2, Phl p 5, and Phl p 6) fused to the PreS carrier protein from hepatitis B, which lacks relevant allergenic activityE10xE10Niederberger, V., Marth, K., Eckl-Dorna, J., Focke-Tejkl, M., Weber, M., Hemmer, W. et al. Skin test evaluation of a novel peptide carrier-based vaccine, BM32, in grass pollen-allergic patients. J Allergy Clin Immunol. 2015; 136: 1101–1103
Abstract | Full Text | Full Text PDF | PubMed | Scopus (26) | Google ScholarSee all References and therefore can be injected into allergic patients without need for updosing.3x3Focke-Tejkl, M., Weber, M., Niespodziana, K., Neubauer, A., Huber, H., Henning, R. et al. Development and characterization of a recombinant, hypoallergenic, peptide-based vaccine for grass pollen allergy. J Allergy Clin Immunol. 2015; 135: 1207–1217 (e1-11)
Abstract | Full Text | Full Text PDF | PubMed | Google ScholarSee all References, 4x4Zieglmayer, P., Focke-Tejkl, M., Schmutz, R., Lemell, P., Zieglmayer, R., Weber, M. et al. Mechanisms, safety and efficacy of a B cell epitope-based vaccine for immunotherapy of grass pollen allergy. EBioMedicine. 2016; 11: 43–57
Abstract | Full Text | Full Text PDF | PubMed | Scopus (43) | Google ScholarSee all References For our immunogenicity studies, we have used a dose (ie, 20 μg of each of the 4BM fusion proteins) that has been safely administered to allergic patients (ClinicalTrials.gov identifiers NCT01445002, NCT01538979, and NCT02643641).4 x4Zieglmayer, P., Focke-Tejkl, M., Schmutz, R., Lemell, P., Zieglmayer, R., Weber, M. et al. Mechanisms, safety and efficacy of a B cell epitope-based vaccine for immunotherapy of grass pollen allergy. EBioMedicine. 2016; 11: 43–57
Abstract | Full Text | Full Text PDF | PubMed | Scopus (43) | Google ScholarSee all ReferencesThe main finding of our current study was that 3 monthly subcutaneous injections of aluminum hydroxide–adsorbed BM32 induced IgG antibody levels to the major grass pollen allergens Phl p 1, Phl p 5, and Phl p 6 in rabbits, which were comparable with natural allergen extract–based registered grass pollen allergy vaccines requiring more than 8 injections (Allergovit grass; Allergopharma, Reinbek, Germany; Alutard SQ grass mix; ALK-Abelló, Hørsholm, Denmark; and Phostal grasses + rye; Stallergenes, Antony, France; see the MethodsMethods section in this article's Online Repository at www.jacionline.org; Fig 1Fig 1), whereas almost no response was observed with Pollinex (Pollinex Quattro Plus grasses + rye; Bencard Allergie GmbH, Munich, Germany), a vaccine based on 4 injections. Importantly, BM32 induced higher levels of Phl p 2–specific IgG antibodies than any of the registered allergen extract–based vaccines (Fig 1Fig 1). We consider this an important finding because it has been shown that group 2 allergens are major grass pollen allergens recognized by more than 60% of patients with grass pollen allergy and, when compared with the other grass pollen allergens by using skin testing, were found to show high allergenic potency.5x5Westritschnig, K., Horak, F., Swoboda, I., Balic, N., Spitzauer, S., Kundi, M. et al. Different allergenic activity of grass pollen allergens revealed by skin testing. Eur J Clin Invest. 2008; 38: 260–267
Crossref | PubMed | Scopus (42) | Google ScholarSee all References Thus our results indicate that it should be possible to build up sufficient levels of grass pollen allergen–specific IgG responses with only few injections (ie, 3-5) of BM32, whereas traditional allergy vaccines require more than double the number of updosing injections. Sublingual treatment even requires daily administration. Therefore we think that the treatment schedules based on BM32 will be more convenient for patients and should increase their compliance. In the IgE inhibition experiments BM32-induced IgG blocked IgE binding to Phl p 2 significantly better than IgG antibodies induced by the commercial vaccines (P = .0008, Fig 2Fig 2). Therefore one might assume that BM32 could be superior to the extract-based vaccines regarding protection of group 2 allergen–induced symptoms. Finally, we also could demonstrate that BM32-induced antibodies inhibited specifically grass pollen allergen–induced T-cell proliferation (see the MethodsMethods section and Fig E3Fig E3 in this article's Online Repository at www.jacionline.org), which suggests that BM32 might also have beneficial effects on T cell–mediated late-phase symptoms of grass pollen allergy. However, a limitation of our study is that we performed the immunogenicity study in naive animals, and therefore there might be differences regarding already sensitized patients.
In summary, our immunogenicity studies indicate that induction of grass pollen allergen–specific protective antibody responses requires considerably fewer injections of BM32 compared with natural allergen extract–based vaccines and that the use of carrier proteins in the vaccine helps to overcome the poor immunogenicity of group 2 allergens. Given the strong reduction of allergenic activity of BM32, one might expect that BM32 represents a grass pollen allergy vaccine that could be superior to allergen extract–based vaccines.
Authors:
Milena Weber Katarzyna Niespodziana Birgit Linhart Angela Neubauer ,
Hans Huber Rainer Henning Rudolf Valenta
　　创建过敏性疾病的科研、科普知识交流平台，为过敏患者提供专业诊断、治疗、预防的共享平台。