CN107635584B - 基因治疗以预防对过敏原的反应 - Google Patents
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Abstract
本发明涉及一种载体,其包含可操作地连接到编码阻断过敏反应的治疗基因的核酸序列的启动子。本发明还涉及包含该载体的组合物以及使用该载体以减少或抑制哺乳动物中对过敏原的免疫应答或过敏反应的方法。
Description
相关申请的交叉引用
本专利申请要求于2016年3月29日提交的美国临时专利申请号62/314,740和于2015年4月9日提交的美国临时专利申请号62/145,035的权益,其通过引用并入本文。
以电子方式提交材料的引用并入
本文通过引用将其全部内容并入的是与本文同时提交并且如下标识的计算机可读核苷酸/氨基酸序列表:于2016年3月29日创建的名为“722135_ST25.TXT”的一个5,718字节ASCII(文本)文件。
发明背景
过敏原在易感个体中引起各种反应,从皮疹到致命的过敏反应。这些反应由与过敏原抗原特异性免疫球蛋白E(IgE)连接的I型超敏反应介导。已经有相当大的兴趣,用中断过敏原特异性IgE诱发过敏反应的疗法来治疗过敏性个体。一种这样的方法是用重组DNA衍生的人源化IgG1κ单克隆抗体奥马珠单抗(其结合人IgE)来治疗。奥马珠单抗抑制IgE与肥大细胞和嗜碱性粒细胞表面上的IgE受体的结合,从而限制了过敏反应介质的释放程度。
使用抗IgE单克隆抗体作为在敏感性个体中针对过敏原诱导的过敏反应的预防性治疗的挑战在于:单次给予奥马珠单抗所提供的保护估计为2至4周。当前治疗的较短半衰期需要至少每月肠胃外给药奥马珠单抗以维持持久的有效治疗。
因此,需要开发替代的组合物和方法来施用IgE特异性抗体并预防性治疗过敏原诱导的过敏反应。本发明提供了此类组合物和方法。根据本文提供的详细描述,本发明的这个优点和其它优点将变得明显。
发明概述
本发明提供了一种载体,其包含可操作地连接到核酸序列的启动子,所述核酸序列编码抗IgE抗体或其抗原结合片段,或编码可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4。本发明还提供了包含所述载体的组合物,以及使用所述载体来抑制或减少哺乳动物中对过敏原的免疫应答或过敏反应的方法。此外,本发明提供了一种提供重组人源化小鼠过敏模型的方法。
附图简述
图1A是用于开发重组人源化小鼠过敏模型的实验方案的示意图。
图1B-1D是描述实验数据的图,其说明了通过ELISA测量的用来自花生过敏性或对照供体的人血单核细胞重建的NOD-scid-IL2Rγnull(NSG)小鼠中总人IgG(图1B)、总人IgE(图1C)和花生特异性IgE(图1D)的水平(平均值±SEM,n=4/组)。
图1E提供了花生提取物攻击后小鼠的图像。左栏-用来自非过敏性供体的单核细胞重建的小鼠,其在花生攻击后表现正常。右栏-用来自花生过敏性供体的单核细胞重建的小鼠,其在花生攻击后1分钟显示出眼/口鼻周围的浮肿、毛发直立和皮毛瘙痒/皱褶。
图1F-1G说明在第5周发生攻击后30分钟,在花生过敏性(n=3)和对照小鼠(n=3)中的过敏反应评分(1至5)(图1F)或血浆组胺水平(图1G)的实验数据图。
图1H是说明小鼠皮肤中被动皮肤过敏反应的图像,其中反应产生图中所示的可见蓝色。在皮内注射50μl来自NSG小鼠的混合血清之前1天对初次实验的(naive)Balb/C小鼠的腹部剃毛,所述NSG小鼠用来自花生过敏性个体或来自对照供体(致敏并在第7.5周时用花生提取物攻击)的血液单核细胞重建。皮内注射该血清24小时后,向小鼠静脉注射100μl0.5%Evan蓝染料和100μg花生提取物的混合物。30分钟后,将小鼠处死,将腹部皮肤翻转,并通过可见的蓝色检查反应。
图2A-2C描述的实验数据说明了奥马珠单抗处理致敏和攻击后的NSG小鼠中的花生抗原诱导的过敏反应的有效性。图2A是显示通过ELISA测量的在处理前1周和用奥马珠单抗处理后1周的游离IgE水平的图(n=9)。图2B是用奥马珠单抗处理的小鼠用花生攻击后呈现正常的图像,图2C是描绘奥马珠单抗介导的被动皮肤过敏反应抑制的图像。
图3A是AAVrh.10抗hIgE载体的示意图,其显示了CMV增强子/鸡β-肌动蛋白(CAG)启动子,抗IgE单克隆抗体奥马珠单抗的重链和轻链,弗林蛋白酶2A切割位点和聚腺苷酸化信号。
图3B是Western印迹的图像,其描述了由AAVrh.10抗hIgE载体编码的抗IgE抗体在HEK 293细胞中的表达。
图3C是实验数据图,其说明了单次静脉内施用AAVrh.10抗hIgE或AAVrh.10IgG对照至NSG小鼠(n=5)后抗IgE抗体的经时持续表达。
图3D是实验数据图,其说明了单次静脉内施用AAVrh.10抗hIgE、AAV9抗hIgE、AAV8抗hIgE或AAVrh.10抗尼古丁(对照)至Balb/C雌性小鼠(n=5/组)后抗IgE抗体的经时持续表达。
图3E是实验数据图,其说明了单次静脉内施用AAVrh.10抗hIgE、AAV9抗hIgE、AAV8抗hIgE或AAVrh.10抗尼古丁(对照)至NSG雌性小鼠(n=5/组)后抗IgE抗体的经时持续表达。
图4A是用于测试NSG小鼠的预防性治疗的治疗方案的示意图。
图4B-4D是实验数据图,其说明了通过ELISA测量的用来自花生过敏性的用AAVrh.10抗hIgE载体或AAVrh.10抗hIgG对照载体处理的人血单核细胞重建的IL2Rγnull小鼠(NSG)或用来自对照供体的人血单核细胞重建的小鼠中的总人IgE(图4B)、总花生特异性IgE(图4C)和游离IgE(图4D)水平(平均值±SEM,图B和C;n=8不具有PN过敏的供体,n=7具有PN过敏的供体+AAVrh.10IgG对照,n=10具有PN过敏的供体+AAVrh.10抗hIgE,图C;n=8无PN过敏的供体,n=7具有PN过敏的供体+AAVrh.10IgG对照,n=10具有PN过敏的供体+AAVrh.10抗hIgE)。
图5A提供了在第6周花生提取物攻击之后的小鼠图像。左-在第3周用对照载体处理的小鼠在花生攻击后显示出眼/口鼻周围的浮肿和毛发直立、皮毛瘙痒/皱褶、行走和呼吸率降低。右-在第3周用AAVrh.10抗hIgE处理的小鼠在花生攻击后表现正常。
图5B-5E是描述AAVrh.10抗hIgE处理后过敏反应变化的图表和图像。图5B描绘了基于累积行进距离的红外光束旷场室评估的自发活动。所示为在第6周评估的载体和对照处理的小鼠中之后30分钟所横穿的距离(n=8不具有PN过敏的供体,n=6具有PN过敏的供体+AAVrh.10IgG对照,n=10具有PN过敏的供体+AAVrh.10抗hIgE)。
图5C描述了在第6周花生攻击后30分钟的过敏反应评分(n=8不具有PN过敏的供体,n=6具有PN过敏的供体+AAVrh.10IgG对照,n=10具有PN过敏的供体+AAVrh.10抗hIgE)。
图5D描绘了在第7周花生攻击后30分钟的血浆组胺水平(n=7不具有PN过敏的供体,n=6具有PN过敏的供体+AAVrh.10IgG对照,n=10具有PN过敏的供体+AAVrh.10抗hIgE)。
图5E描绘了AAVrh.10抗hIgE介导的被动皮肤过敏反应的抑制:左栏-花生提取物诱导的、由来自花生过敏性供体血清的花生特异性IgE介导的被动皮肤过敏反应;右栏-花生提取物诱导的、由来自人源化花生过敏性NSG小鼠的混合血清的花生特异性IgE介导的被动皮肤过敏反应,所述小鼠使用与左栏中相同的供体重建,但在致敏前3周用AAVrh.10抗hIgE进行预防性处理。与AAVrh.10IgG对照相比,来自AAVrh.10抗hIgE处理的小鼠的血清阻断了花生诱导的花生特异性IgE介导的被动皮肤过敏反应。
图6A是用于在NSG小鼠中测试对花生诱导的过敏反应的治疗性处理的治疗方案的示意图。图6B-6D是实验数据图,其说明了通过ELISA测量的用来自花生过敏性个体的人血单核细胞重建的、用AAVrh.10抗hIgE载体或AAVrh.10抗hIgG对照处理的NSG小鼠中的总人IgE(图6B)、总花生特异性IgE(图6C)和游离IgE(图6D)的水平(平均值±SEM)。
图7A提供了在第10周花生提取物攻击后小鼠的图像:左栏-用奥马珠单抗处理的小鼠在花生攻击后表现出眼/口鼻周围浮肿和毛发直立、皮毛瘙痒/皱褶、行走和呼吸率降低;右栏-在第10周(治疗后5周)用AAVrh.10抗hIgE处理的小鼠在花生攻击后表现正常。
图7B-7E是描述AAVrh.10抗hIgE处理后过敏反应变化的图表和图像。图7B描绘了基于累积行进距离的红外光束旷场盒评估的自发行动,所述评估在对载体、对照处理的和奥马珠单抗小鼠的花生攻击后30分钟开始。所示数据为第7周(即,治疗后2周)和第10周(即,治疗后5周)的接下来30分钟所横穿的距离。第7周的数据:对于AAVrh.10抗hIgE n=10,对于奥马珠单抗n=9,对于AAVrh.10IgG对照n=7,第10周的数据:对于AAVrh.10抗hIgEn=9,对于奥马珠单抗n=5,且对于AAVrh.10IgG对照n=0。
图7C描述了花生攻击后30分钟的过敏反应评分。所示为第7周(即,治疗后2周)和第10周(即,治疗后5周)的数据。第7周数据:对于AAVrh.10抗hIgE n=10,对于奥马珠单抗n=9,对于AAVrh.10IgG对照n=7。第10周数据:对于AAVrh.10抗hIgE n=9,对于奥马珠单抗n=5,且对于AAVrh.10IgG对照n=0。
图7D描述了花生攻击后30分钟的血浆组胺水平。所示为第6周(即,治疗后1周)和第9周(即,治疗后4周)的数据。第6周数据:对于AAVrh.10抗hIgE n=10,对于奥马珠单抗n=9,且对于AAVrh.10IgG对照n=7。第9周数据:对于AAVrh.10抗hIgE n=9,对于奥马珠单抗n=6,且对于AAVrh.10IgG对照n=0。
图7E描绘了AAVrh.10抗hIgE介导的被动皮肤过敏反应的抑制。上栏-第7周(治疗后2周)花生提取物诱导的、由来自花生过敏性供体(但并非来自非过敏性对照)的血清的花生特异性IgE介导的被动皮肤过敏反应。下栏-第10周(即,治疗后5周)持续表达的AAVrh.10抗hIgE阻断染料的外渗,而5周前一次注射的奥马珠单抗不再是保护性的。
图8是描述用AAVrh.10抗-hIgE、单独的奥马珠单抗或对照载体处理后小鼠存活的图表。生存期和治疗类型按照治疗后天数显示。
发明详述
本发明提供了基因治疗载体和使用该基因治疗载体来提供治疗性转基因的持续表达以抑制或减少哺乳动物中对过敏原的免疫应答或过敏反应的方法。该载体包含、基本由、或由可操作地连接到核酸序列的启动子组成,该核酸序列编码抗IgE抗体或其抗原结合片段,或编码可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4。该载体可以包含不会实质影响载体的另外的组分(例如,促进体外载体操作的遗传元件,例如聚(A)序列或限制酶位点)。然而,在一些实施方式中,载体不包含任何另外的组分(即对载体而言不是内源性的组分,并且不需要其影响核酸序列的表达从而提供抗体)。
本发明的载体可以包含、基本由、或由本领域已知的任何基因转移载体组成。此类载体的实例包括腺相关病毒(AAV)载体、腺病毒载体、慢病毒载体、逆转录病毒载体和质粒。在一个优选的实施方式中,载体是AAV载体。
腺相关病毒是细小病毒科(Parvoviridae)的成员,并且包含少于约5,000个核苷酸的线性单链DNA基因组。AAV需要与辅助病毒(即,腺病毒或疱疹病毒)共同感染,或辅助基因的表达,以有效复制。用于施用治疗性核酸的AAV载体通常具有约96%的亲本基因组缺失,使得仅保留含有用于DNA复制和包装的识别信号的末端重复(ITR)。这消除了由于病毒基因表达所引起的免疫或毒副作用。另外,如果需要,将特异性AAV蛋白递送至产生细胞使得能够将包含AAV ITR的AAV载体整合至细胞基因组的特定区域中(参见例如美国专利6,342,390和6,821,511)。包含整合的AAV基因组的宿主细胞在细胞生长或形态学上没有变化(参见例如美国专利4,797,368)。
AAV ITR侧接非结构性复制(Rep)蛋白和结构衣壳(Cap)蛋白(也称为病毒体蛋白(VP))的独特编码核苷酸序列。末端145个核苷酸是自身互补的并且有序排列(organized),从而可以形成能量上稳定的形成T形发夹的分子内双链体。这些发夹结构通过作为细胞DNA聚合酶复合物的引物而起到病毒DNA复制起点的作用。Rep基因编码Rep蛋白Rep78、Rep68、Rep52和Rep40。Rep78和Rep68自p5启动子转录,而Rep52和Rep40自p19启动子转录。Rep78和Rep68蛋白是在生产性复制期间执行解旋酶和切口酶功能以允许AAV末端解析的多功能DNA结合蛋白(参见例如Im等人,Cell,61:447-57(1990))。这些蛋白质还调节自内源性AAV启动子和辅助病毒内的启动子的转录(参见例如Pereira等人,J.Virol.,71:1079-1088(1997))。其它Rep蛋白修饰Rep78和Rep68的功能。Cap基因编码衣壳蛋白VP1、VP2和VP3。Cap基因自p40启动子转录。
本发明的AAV载体可以使用本领域已知的任何AAV血清型来产生。已经从腺病毒原种或者人或非人灵长类动物组织中分离了若干种AAV血清型和超过100种AAV变体(综述于例如Wu等人,Molecular Therapy,14(3):316-327(2006))。通常,AAV血清型在核酸序列和氨基酸序列水平上具有显著同源的基因组序列,使得不同血清型具有相同的一组遗传功能,产生在物理和功能上基本等同的病毒颗粒,并通过实际上相同的机制复制和组装。AAV血清型1-6和7-9被定义为“真”血清型,因为它们不与特异于所有其它现有的并已被表征的血清型的中和血清有效地交叉反应。相反,AAV血清型6、10(也称为Rh10)和11被认为是“变体”血清型,因为它们不符合“真”血清型的定义。AAV血清型2(AAV2)由于缺乏致病性、广泛的感染性和建立长期转基因表达的能力而广泛用于基因治疗应用(参见例如Carter,B.J.,Hum.Gene Ther.,16:541-550(2005);和Wu等人,同上)。各种AAV血清型的基因组序列及其比较公开于例如GenBank登录号U89790、J01901、AF043303和AF085716;Chiorini等人,J.Virol.,71:6823-33(1997);Srivastava等人,J.Virol.,45:555-64(1983);Chiorini等人,J.Virol.,73:1309-1319(1999);Rutledge等人,J.Virol.,72:309-319(1998);和Wu等人,J.Virol.,74:8635-47(2000))中。
AAV rep和ITR序列在大多数AAV血清型中特别保守。例如,据报道,AAV2、AAV3A、AAV3B、AAV4和AAV6的Rep78蛋白约89-93%相同(参见Bantel-Schaal等人,J.Vitol.,73(2):939-947(1999))。已报道AAV血清型2、3A、3B和6在基因组水平上具有约82%的总核苷酸序列同一性(Bantel-Schaal等人,同上)。此外,已知许多AAV血清型的rep序列和ITR在哺乳动物细胞中产生AAV颗粒期间有效地交叉补充(即功能上取代)来自其它血清型的相应序列。
通常,在不同AAV血清型之间,确定AAV颗粒的细胞嗜性的cap蛋白以及相关的cap蛋白编码序列比Rep基因保守性低得多。鉴于Rep和ITR序列交叉互补其它血清型的相应序列的能力,AAV载体可以包含血清型的混合物,从而成为“嵌合”或“假型”AAV载体。嵌合AAV载体通常包含源自两种或更多种(例如,2、3、4种等)不同AAV血清型的AAV衣壳蛋白。相比之下,假型AAV载体包含一种AAV血清型的一种或多种ITR,但包装到另一种AAV血清型的衣壳中。嵌合和假型AAV载体进一步描述于例如美国专利6,723,551;Flotte,Mol.Ther.,13(1):1-2(2006);Gao等人,J.Virol.,78:6381-6388(2004);Gao等人,Proc.Natl.Acad.Sci.USA,99:11854-11859(2002);De等人,Mol.Ther.,13:67-76(2006);和Gao等人,Mol.Ther.,13:77-87(2006)中。
在一个实施方式中,使用感染人的AAV(例如,AAV2)产生AAV载体。在一个优选的实施方式中,使用感染人的AAV产生的AAV载体是AAV8或AAV9。或者,使用感染非人类灵长类例如大猩猩(例如,黑猩猩)、旧世界猴(例如,恒河猴)和新世界猴(例如,狨猴)的AAV产生AAV载体。优选地,使用AAV产生AAV载体,所述AAV是感染人的AAV假型包装的感染非人灵长类的AAV。此类假型AAV载体的实例公开于例如Cearley等人,Molecular Therapy,13:528-537(2006)中。在一个实施方式中,可以产生AAV载体,所述AAV载体包含来自感染恒河猴的AAV的衣壳蛋白,并且被AAV2反向末端重复(ITR)假型包装。
在一个特别优选的实施方式中,本发明的AAV载体包含来自感染恒河猴的AAV10的衣壳蛋白(也称为“AAVrh.10”),并被AAV2的ITR假型包装(参见例如Watanabe等人,GeneTher.,17(8):1042-1051(2010);和Mao等人,Hum.Gene Therapy,22:1525-1535(2011))。
本发明的载体包含可操作地连接到核酸序列的启动子,该核酸序列编码抗IgE抗体或其抗原结合片段,或编码可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4。DNA区域在功能上彼此相关时是“可操作地连接”的。如果启动子控制编码序列的转录,则启动子“可操作地连接”到该序列。
“启动子”是启动特定基因转录的DNA区域。来自各种不同来源的大量启动子在本领域是众所周知的。启动子的代表性来源包括例如病毒、哺乳动物、昆虫、植物、酵母和细菌,并且来自这些来源的合适的启动子容易获得,或者可以基于公共可获得的序列(例如,来自诸如ATCC的保藏处以及其他商业或个人来源)而合成得到。启动子可以是单向的(即,以一个方向启动转录)或双向的(即,以3′或5′方向启动转录)。
本发明载体的启动子可以包含、基本由或由本领域已知的任何启动子组成。此类启动子类型的实例包括组成型活性启动子(例如,人β-肌动蛋白、鸡β-肌动蛋白、巨细胞病毒(CMV)和SV40)、细胞类型特异性启动子(例如,CD19基因启动子、CaMKIIa和UAS)或诱导型启动子(例如,Tet系统(美国专利5,464,758和5,814,618)、蜕皮激素诱导型系统(No等人,Proc.Natl.Acad.Sci.,93:3346-3351(1996))、T-REXTM系统(Invitrogen,Carlsbad,CA)、Cre-ERT他莫昔芬诱导型重组酶系统(Indra等人,Nuc.Acid.Res.,27:4324-4327(1999);Nuc.Acid.Res.,28:e99(2000);美国专利7,112,715;和Kramer&Fussenegger,MethodsMol.Biol.,308:123-144(2005))和LACSWITCHTM系统(Stratagene,San Diego,CA))。
在本发明的一个优选实施方式中,启动子是组成型活性启动子、诱导型启动子或细胞类型特异性启动子。启动子的一个例子是鸡β-肌动蛋白启动子。
“核酸序列”旨在涵盖DNA或RNA的聚合物,即多核苷酸,其可以是单链或双链的,并且可以含有非天然的或改变的核苷酸。如本文使用的术语“核酸”和“多核苷酸”是指任何长度的核糖核苷酸(RNA)或脱氧核糖核苷酸(DNA)的核苷酸聚合形式。这些术语是指分子的一级结构,因此包括双链和单链DNA,以及双链和单链RNA。该术语包括作为等同物的由核苷酸类似物和经修饰多核苷酸(例如,但不限于甲基化和/或封端的多核苷酸)制备的RNA或DNA的类似物。
可操作地连接到启动子的核酸序列可以包含编码阻断过敏反应的治疗性基因的任何核酸序列。核酸序列优选编码抗IgE抗体或其抗原结合片段、可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞,IL-13或IL-4。核酸序列还可以编码融合蛋白,其包含活性蛋白(例如,可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13、IL-4)或阻断过敏反应的任何治疗性基因,和通常是蛋白质的第二部分,其改善活性蛋白的性质(例如,疗效、溶解性或半衰期)。第二部分的实例是本领域已知的,并且包括例如免疫球蛋白的Fc结构域和聚乙二醇(PEG)。在一个实施方式中,可操作地连接到启动子的核酸序列仅编码抗IgE抗体或其抗原结合片段。
本领域普通技术人员将会理解,抗体由四条多肽组成:两个相同拷贝的重(H)链多肽和两个拷贝的轻(L)链多肽。每条重链含有一个N末端可变(VH)区和三个C末端恒定(CH1、CH2和CH3)区,每条轻链含有一个N末端可变(VL)区和一个C末端恒定(CL)区。每对轻链和重链的可变区形成抗体的抗原结合位点。本发明的载体可以包含一个或多个核酸序列,每个核酸序列编码抗IgE抗体的一个或多个重链和/或轻链多肽。在这方面,本发明的载体可以包含编码抗IgE抗体的两个重链多肽和两个轻链多肽的单个核酸序列。或者,本发明的载体可以包含编码抗IgE抗体的两个重链多肽的第一核酸序列,和编码抗IgE抗体的两个轻链多肽的第二核酸序列。在又一个实施方式中,本发明的载体可以包含编码抗IgE抗体的第一重链多肽的第一核酸序列、编码抗IgE抗体的第二重链多肽的第二核酸序列、编码抗IgE抗体的第一轻链多肽的第三核酸序列和编码抗IgE抗体的第二轻链多肽的第四核酸序列。
在另一实施方式中,载体可以包含编码抗IgE抗体的抗原结合片段(也称为“抗体片段”)的核酸序列。术语“抗原结合片段”是指保留特异性结合抗原(例如,免疫球蛋白E)的能力的一种或多种抗体片段(通常参见Holliger等人,Nat.Biotech.,23(9):1126-1129(2005))。抗原结合片段的实例包括但不限于(i)Fab片段,其是由VL、VH、CL和CH1结构域组成的单价片段;(ii)F(ab’)2片段,其是包含通过铰链区处的二硫键连接的两个Fab片段的二价片段;和(iii)由抗体单臂的VL和VH结构域组成的Fv片段。在一个实施方式中,载体可以包含编码抗IgE抗体的Fab片段的核酸序列。
核酸序列可以编码本领域已知的任何抗IgE抗体或其抗原结合片段。在一个实施方式中,核酸序列可以编码抗IgE抗体或其抗原结合片段,其包含含有三个互补决定区(CDR)的重链多肽,其中CDR-H1包含SEQ ID NO:1的核酸序列,CDR-H2包含SEQ ID NO:2的核酸序列,CDR-H3包含SEQ ID NO:3的核酸序列,和包含三个CDR的轻链多肽。其中CDR-L1包含SEQ ID NO:4的核酸序列,CDR-L2包含SEQ ID NO:5的核酸序列,CDR-L3包含SEQ ID NO:6的核酸序列。
在另一个实施方式中,核酸序列可以编码抗IgE抗体或其抗原结合片段,其包含含有SEQ ID NO:7的重链可变区和含有SEQ ID NO:8的轻链可变区。
在另一个实施方式中,核酸序列编码包含SEQ ID NO:9的抗IgE抗体或其抗原结合片段。
在另一个实施方式中,核酸序列可以编码抗IgE抗体或其抗原结合片段,其包含高亲和力IgE结合单克隆抗体奥马珠单抗(参见例如美国专利6,682,735)或其抗原结合片段、或与奥马珠单抗结合相同表位的抗IgE抗体或抗体片段。在这方面,本发明的载体可以包含编码奥马珠单抗的全长重链和轻链多肽(例如,分别为SEQ ID NO:10和SEQ ID NO:11)的核酸序列。
抗体或其抗原结合片段可以通过任何方式获得,包括经由体外来源(例如,重组产生抗体的杂交瘤或细胞系)和体内来源(例如,啮齿动物)。用于产生抗体的方法是本领域已知的并且描述于例如和Milstein,Eur.J.Immunol.,5:511-519(1976);Harlow和Lane(编),Antibodies:A Laboratory Manual,CSH Press(1988);和C.A.Janeway等人(编),Immunobiology,第5版,Garland Publishing,New York,NY(2001))中。在某些实施方式中,可以使用转基因动物(例如,小鼠)产生人抗体或嵌合抗体,所述转基因动物中一个或多个内源性免疫球蛋白基因被一个或多个人免疫球蛋白基因所替代。其中内源抗体基因被人抗体基因有效替代的转基因小鼠的实例包括但不限于HUMAB-MOUSETM、Kirin TC MOUSETM和KM-MOUSETM(参见例如Lonberg,Nat.Biotechnol.,23(9):1117-25(2005)和Lonberg,Handb.Exp.Pharmacol.,181:69-97(2008))。
编码抗IgE抗体或其抗原结合片段的核酸序列可以使用本领域已知的方法产生。例如,核酸序列、多肽和蛋白质可以使用标准重组DNA方法学重组产生(参见例如Sambrook等人,Molecular Cloning:ALaboratory Manual,第3版,Cold Spring Harbor Press,ColdSpring Harbor,NY,2001;和Ausubel等人,Current Protocols in Molecular Biology,Greene Publishing Associates and John Wiley&Sons,NY,1994)。另外,编码抗IgE抗体或其抗原结合片段的合成产生的核酸序列可以从诸如细菌、昆虫或哺乳动物(例如,大鼠、人等)的来源分离和/或纯化。分离和纯化的方法是本领域众所周知的。或者,本文所述的核酸序列可以商业合成。在这方面,核酸序列可以是合成的、重组的、分离的和/或纯化的。
除了可操作地连接到编码抗IgE抗体或其抗原结合片段、可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13、IL-4的核酸序列的启动子之外,载体可以包含提供核酸序列在宿主细胞中表达的另外的表达控制序列,例如增强子、多聚腺苷酸化信号、转录终止子、内部核糖体进入位点(IRES)等。示例性的表达控制序列是本领域已知的,并且描述在例如Goeddel,Gene Expression Technology:Methods in Enzymology,Vol.185,AcademicPress,San Diego,CA.(1990)中。
如本文使用的术语“增强子”是指增加例如与其可操作地连接的核酸序列的转录的DNA序列。增强子可以位于距核酸序列编码区数千个碱基处,并且可以介导调节因子的结合、DNA甲基化的模式或DNA结构的变化。来自各种不同来源的大量增强子是本领域所众所周知的,并且可以作为克隆的多核苷酸或在克隆的多核苷酸内(例如,来自诸如ATCC的保藏处以及其它商业或个体来源)获得。包含启动子(例如,常用的CMV启动子)的许多多核苷酸也包含增强子序列。增强子可以位于编码序列的上游、内部或下游。编码抗IgE抗体或其抗原结合片段、可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4的核酸序列可以可操作地连接到CMV增强子/鸡β-肌动蛋白启动子(也称为“CAG启动子”)(参见例如Niwa等人,Gene,108:193-199(1991);Daly等人,Proc.Natl.Acad.Sci.U.S.A.,96:2296-2300(1999);和Sondhi等人,Mol.Ther.,15:481-491(2007))。
本发明提供了包含、基本由或由上述载体和药学上可接受的(例如,生理学上可接受的)运载体组成的组合物。当组合物基本上由本发明的载体和药学上可接受的运载体组成时,可以包含不会实质上影响组合物的另外组分(例如,佐剂、缓冲剂、稳定剂、抗炎剂、增溶剂、防腐剂等)。当组合物由本发明的载体和药学上可接受的运载体组成时,组合物不包含任何另外的组分。在本发明的背景下可以使用任何合适的运载体,并且此类运载体在本领域中是众所周知的。运载体的选择将部分地由组合物可施用的特定部位和用于施用组合物的具体方法来确定。除了本文所述的载体外,组合物任选地可以是无菌的。可以将组合物冷冻或冻干以便储存,并在使用之前在合适的无菌运载体中重构。组合物可根据例如Remington:The Science and Practice of Pharmacy,第21版,Lippincott Williams&Wilkins,Philadelphia,PA(2001)中所述的常规技术来产生。
适用于组合物的制剂包括水性和非水性溶液、等渗无菌溶液,其可包含抗氧化剂、缓冲剂和抑菌剂,以及水性和非水性无菌混悬液,其可包含悬浮剂、增溶剂、增稠剂、稳定剂和防腐剂。制剂可以以存在于单位剂量或多剂量密封容器(例如,安瓿和小瓶)中,并且可以贮存在冷冻干燥(冻干)条件下,仅需在即用前加入无菌液体运载体(例如,水)。即时溶液和混悬液可以由前述种类的无菌粉末、颗粒和片剂制备。优选地,载体是缓冲盐溶液。更优选地,将本发明的载体以经配制组合物形式使用,以保护其在施用之前免受损害。例如,可以配制组合物以减少载体在用于制备、存储或施用载体的装置(例如,玻璃器皿、注射器或针)上的损失。可以配制组合物以降低载体的光敏感性和/或温度敏感性。为此,组合物优选包含药学上可接受的液体运载体,例如上述那些,和选自聚山梨醇酯80、L-精氨酸、聚乙烯吡咯烷酮、海藻糖及其组合的稳定剂。使用此组合物将延长载体的保质期、促进施用、并增加本发明方法的效率。含有载体的组合物的制剂进一步描述于例如Wright等人,Curr.Opin.DrugDiscov.Devel.,6(2):174-178(2003)和Molecular Therapy,12:171-178(2005)中。
组合物也可以经配制以增强转导效率。此外,本领域普通技术人员将会理解,本发明的载体可以与其它治疗剂或生物活性剂一起存在于组合物中。例如,控制炎症的因子例如布洛芬或类固醇可以是组合物的一部分,以减少与体内施用载体相关的肿胀和炎症。可存在抗菌素,即杀微生物剂和杀真菌剂,以治疗现有感染和/或降低未来感染的风险,例如与转基因过程相关的感染。
本发明提供了抑制或减少哺乳动物中对过敏原的免疫应答或过敏反应的方法,其包括对哺乳动物施用本发明的载体,由此表达核酸以产生抑制或减少免疫应答的蛋白质。在一个优选的实施方式中,哺乳动物是人。
抑制或减少对过敏原的免疫应答或过敏反应包括对过敏原的任何生理反应的任何程度的改善。生理反应的非限制性实例包括荨麻疹、皮疹、粘液产生和过敏反应。在一个优选的实施方式中,通过该方法减少或抑制的免疫应答或过敏反应是过敏反应(anaphylaxis)。
本发明的过敏原可以是在哺乳动物中引起过敏反应的任何过敏原。可以通过本发明的方法治疗的过敏原的非限制性实例包括:
食物过敏原,例如花生、树坚果(榛子、扁桃仁、腰果、澳洲坚果、开心果、松子、核桃、巴西果、栗子、山核桃)、鱼类/甲壳类/贝类(鳎目鱼、鱿鱼、鲭鱼、鳕鱼、蓝贻贝、铡刀鱼、梭子鱼、大比目鱼、金枪鱼、鲭鱼、鲑鱼、鳟鱼、鳕鱼、凤尾鱼、狭鳕鱼、鲶鱼、红鲷鱼、鲱鱼、比目鱼、鲑鱼、鳟鱼、剑鱼、白鱼、牡蛎、扇贝、沙丁鱼、小龙虾、黑线鳕、罗非鱼、螃蟹、虾、蛤蜊、鲈鱼、章鱼)、大豆、牛奶/乳制品(山羊奶、牛奶等)、小麦、面筋、亚硫酸盐、芝麻、大蒜、燕麦、乳清、莳萝、罗勒、百里香、山药、鼠尾草、石灰、丁香、薄荷、蜂蜜、牛至、肉豆蔻、甜菜、罂粟籽、鸢尾根、生姜、黄瓜、芦笋、蔓越莓、西葫芦、覆盆子、红醋栗、迷迭香、卵清蛋白、洋蓟、黑豆、枯茗籽、油桃、苹果、李子、香蕉、姜黄、柑橘、藜麦,南瓜、黑橄榄、青橄榄、真菌/霉菌(奶酪霉菌/食品霉菌)、橙、玉米、西瓜、胡萝卜、马铃薯、利马豆、白豆、豌豆、胡椒、茴香、西葫芦、葵花籽、绿豆、葛缕子籽、小豆蔻籽、角豆树(胶)/刺槐豆,明胶(猪、牛、鱼)、南瓜籽、亚麻仁/亚麻籽、香菜/芫荽、黑莓、胭脂树籽、普通小米、花椰菜、菜籽油、鸡豆(鹰嘴豆)、葡萄、番茄、猕猴桃、木瓜、芹菜、鳄梨、荞麦、α-gal、大米、巧克力、鸡肉、火鸡、羊肉、海军豆、黑麦、大麦、酪蛋白、卷心菜、生菜、胡椒、牛肉/猪肉、芒果、梨、菠菜、蛋白、蛋黄、整蛋、木瓜、椰子、杏、蓝莓、蜜瓜、甜瓜、哈密瓜、芥末、茶、香草、柠檬、酸橙、西兰花、肉桂、洋葱、菠萝、大蒜、葡萄柚、小扁豆、麦芽、咖啡、蘑菇、墨西哥辣椒、可可、食品添加剂(面包酵母、抗坏血酸、阿斯巴甜、硝酸盐、瓜尔胶、MSG、卡拉胶);
药物例如β内酰胺抗生素(青霉素、阿莫西林、氨苄青霉素、青霉素G、青霉素V等、头孢菌素、单环胺、碳青霉烯类)、非β内酰胺抗生素、抗分枝杆菌药物、糖尿病药物、癌症化疗药物、HIV药物、用于自身免疫性疾病的免疫调节剂、用于皮肤病学疾病的修饰药物、围手术期试剂、阿片类、皮质类固醇、鱼精蛋白、肝素(抗凝血剂)、局部麻醉剂、放射造影剂、阿司匹林和非甾体抗炎药(NSAID)、血管紧张素转化酶(ACE)抑制剂、生物修饰剂、细胞因子、抗TNF药物、单克隆抗体、抗癌单克隆抗体、补充药物、抗癫痫药物;
环境过敏原例如树花粉、猫(皮屑)、狗(皮屑)、豚鼠、鸭毛、鸡毛、鹅毛、马(毛/皮屑)、豚鼠(上皮)、猪/猪(上皮)、山羊上皮、仓鼠(上皮)、小鼠(上皮)、鸟粪/粪便、昆虫/毒液(蜜蜂、白脸黄蜂、胡蜂、黄脸黄蜂、小黄蜂、火蚁、蚂蚁等)、霉菌/真菌、尘螨、房尘、乳胶、草、螨、杂草、树、蟑螂;和
其它常见的过敏原例如精液/精液液体,血液和血液制品。
优选地,过敏原是食物过敏原(例如,虾或海鲜、花生或树坚果)、花粉、尘螨或昆虫毒液,例如蜂螫刺毒。
可以使用任何施用途径将组合物递送至哺乳动物。事实上,尽管可以使用多于一种途径来施用组合物,但是特定的途径可以提供比另一种途径更直接和更有效的反应。优选地,组合物通过肌内注射施用。也可以将一定剂量的组合物施用或滴注到体腔中、通过皮肤吸收(例如,通过透皮贴剂)、吸入、摄入、局部施用于组织、或通过肠胃外施用,例如静脉内、腹膜内、口内、皮内、皮下或动脉内施用。
组合物可以在允许受控或持续释放的装置中或其上施用,例如海绵、生物相容性网状物、机械储库或机械植入物。植入物(参见例如美国专利5,443,505)、装置(参见例如美国专利4,863,457),例如可植入装置,例如机械储库或植入物或由聚合物组合物构成的装置,对于AAV载体的施用是特别有用的。组合物还可以以持续释放制剂的形式施用(参见例如美国专利5,378,475),其包含例如凝胶泡沫、透明质酸、明胶、硫酸软骨素、聚磷酸酯如对苯二甲酸双-2-羟乙酯(BHET)和/或聚乳酸-乙醇酸。
向哺乳动物施用的组合物中的载体剂量将取决于许多因素,包括哺乳动物的大小(质量)、任何副作用的程度、特定的施用途径等。优选地,本发明的方法包括施用“治疗有效量”的本文所述的包含本发明载体的组合物。“治疗有效量”是指在必要的剂量和时间段内有效实现所需的治疗结果的量。治疗有效量可以根据诸如个体的过敏原敏感程度、年龄、性别和体重以及载体在个体中引起期望的反应的能力等因素而变化。
在另一个实施方式中,本发明的方法可以包括施用“预防有效量”的包含本发明载体的组合物。“预防有效量”是指在必要的剂量和时间段内有效实现所需的预防结果(例如,预防免疫应答或过敏反应)的量。需要预防性施用的受试者可以通过本领域已知的常规过敏试验容易地确定。另外,可以针对将来的过敏反应预防性地治疗具有先前过敏反应的受试者。
编码抗IgE抗体(或可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4)的载体可以在治疗性或预防性治疗期间多次施用和/或采用多种施用途径,例如肌内和皮下,以确保细胞充分暴露于组合物。例如,组合物可以在治疗性或预防性治疗期间施用于哺乳动物两次或更多次(例如,2、3、4、5、6、6、8、9或10次或更多次)。然而,根据本发明的优选方面,本文所述的载体(或包含该载体的组合物)的单次施用足以提供抗IgE抗体(或可溶性IgE受体、嗜酸性粒细胞、嗜碱性粒细胞、IL-13或IL-4)在哺乳动物中治疗或预防水平下的延长表达,与在没有治疗的情况下的免疫应答或过敏反应相比足以抑制或减少对过敏原的免疫应答或过敏反应,且具有很小的副作用。在一些实施方式中,表达水平足以抑制或减少对治疗之间多次暴露于过敏原(例如,暴露于过敏原2次或更多次、3次或更多次、5次或更多次、或甚至10次或更多次)的免疫反应或过敏反应。优选地,在施用载体或包含其的组合物之后,治疗水平在哺乳动物中表达约30天或更多(例如,约45天或更多、约60天或更多、约75天或更多、约90天或更多、约4个月或更多、约6个月或更多、约10个月或更多、或甚至约12个月或更多)。因此,在一些实施方式中,该方法包括向哺乳动物施用载体在约30天内不超过一次、在约45天内不超过一次、在约60天内不超过一次、在约75内不超过一次天、或甚至在约90天内不超过一次(例如,在约4个月、约5个月、约6个月、约10个月或约12个月内不超过一次)。
达到特定的治疗或预防效果(即,减少或抑制过敏反应)所需的组合物中载体的剂量通常以载体基因组拷贝/细胞(gc/细胞)或载体基因组拷贝/每千克体重(gc/kg)的单位施用。本领域普通技术人员可以基于这些和本领域公知的其他因素容易地确定合适的用于治疗具有特定免疫应答的患者的载体剂量范围。
本发明还提供了提供重组人源化小鼠过敏模型的方法,其包括将来自具有过敏的人受试者的外周血单核细胞(PBMC)递送入免疫缺陷小鼠中。在一个优选的实施方式中,血单核细胞来自具有任何过敏或过敏反应临床病史的人受试者。过敏可以是本文先前公开的任何过敏原,优选食物过敏原、花粉、尘螨、昆虫毒液、花生、树坚果或蜂螯毒液。在一个更优选的实施方式中,人受试者具有花生过敏和过敏反应临床病史。可以通过任何合适的方法将PBMC细胞递送入免疫缺陷小鼠中,例如通过注射(例如,腹膜内或静脉内注射)。PBMC可以任选地与相关抗原共施用于小鼠。在一个相关的方面中,本发明还提供了适合用作过敏模型的人源化免疫缺陷小鼠,其中该小鼠包含来自对过敏原过敏的人受试者的PBMC,并且当暴露于过敏原时该小鼠表现出免疫应答或过敏反应。PBMC可来自对如前文所述的任何过敏原过敏的人受试者,所述过敏原例如食物过敏原、花粉、尘螨、昆虫毒液、花生、树坚果或蜜蜂螯毒液。在一个优选的实施方式中,PBMC来自具有花生过敏和过敏反应临床病史的人受试者。
以下实施例进一步说明本发明,但是当然不应将其解释为以任何方式限制其范围。
实施例1
本实施例证实了重组人源化小鼠花生过敏模型的开发。
从具有对花生过敏的供体或非过敏性健康对照受试者获得肝素化血液。通过检测供体血清中的过敏原特异性IgE(ImmunoCAP特异性IgE血液测试;Phadia AB,Uppsala,瑞典)而记录特异性敏化。通过使用Ficoll-Paque密度离心(Paque Plus,SigmaAldrich,St Louis,MO)从肝素化血液中分离血液单核细胞。在用人细胞重建之前,通过ELISA确认小鼠血清中不存在可检测的人IgG。
向6-8周龄的NOD-scidIL@Rgammanull(NSG)小鼠施用通过Ficoll-Paque方法分离的细胞。每只动物腹膜内接受在RPMI(Sigma Aldrich,St Louis,MO)中的3x107个血液单核细胞,其与100μg粗花生在总体积为200μl的0.9%氯化钠中混合,分成两个单独的注射部位(每个100μl)。
通过将25g磨碎的花生与250ml 20mM Tris缓冲液(pH7.2)混合,在施用至小鼠的同一天制备来自经烘焙无盐花生(Arachishypogaea;Hampton Farms;Severn,NC)的蛋白质提取物。2小时后,23℃,收集含水部分,随后离心除去残留的痕量脂肪和不溶性颗粒。使用Bradford分析以牛血清白蛋白作为标准测定蛋白质浓度。
小鼠在第0至4周每周一次通过腹膜内注射100μg粗花生提取物来进行致敏。然后,在第5至10周,通过使用弯曲的20号针头(图1A)用300μg粗花生提取物灌胃来攻击小鼠,并在过敏反应迹象的程序后观察多至4小时。为了最大限度地使花生抗原穿过胃粘膜吸收,所有小鼠在花生攻击前禁食8小时。评估小鼠的总人IgG(图1B)、总人IgE(图1C)、总小鼠IgE、花生特异性(PN特异性)人IgE(图1D)、过敏症状(图1E)、过敏反应评分图(1F)、血浆组胺(图1G)和被动皮肤过敏反应(图1H)。
结果显示用具有或不具有花生过敏的供体的单核细胞重建的小鼠在重建后具有增加的人IgG水平(图1B)。与之相比,在花生致敏后,只有用来自花生过敏受试者的单核细胞重建的小鼠表达总人IgE和花生特异性IgE(图1C和1D)。重要的是,用来自花生过敏供体的血液单核细胞重建的小鼠显示与过敏反应相关的临床表型。小鼠表现出眼和口鼻周围浮肿、毛发直立、皮毛瘙痒/皱褶、步行和呼吸频率下降(图1E),过敏反应评分为2±1(图1F)。这些临床特征在接受来自非过敏性个体的血液单核细胞的动物中未观察到。当与用来自非花生过敏性、非特应性供体的血液单核细胞重建的小鼠比较时,用来自花生过敏性供体的血液单核细胞重建并随后用粗花生提取物攻击的小鼠中的组胺水平在花生攻击后显示出升高的组胺水平(图1G)。
总之,来自这些研究的结果显示用来自花生过敏性供体的血液单核细胞重建的小鼠显示出了与过敏反应相关的表型,而在接受来自非过敏性供体的血液单核细胞的动物中没有观察到这些特征。
为了证明在重建的NOD-scid IL2Rγnull小鼠中的花生诱导的过敏反应是由人IgE介导的,用250μg奥马珠单抗(Novartis,Huningue,France)单次处理重建小鼠的亚组,该剂量是基于如在其它小鼠研究中已经使用的基于体重的剂量。在治疗前和治疗1周后评估小鼠的IgE,并在治疗2周后对小鼠的过敏反应特征和被动皮肤过敏反应进行身体评估。
结果显示,与治疗前一周的水平相比,在给药奥马珠单抗1周后,用奥马珠单抗治疗的小鼠在过敏症状的第一迹象后具有显著更低的(p<0.001)游离IgE水平(图2A)。在花生攻击后,奥马珠单抗小鼠表现正常;奥马珠单抗治疗后2周(比较图2B与图1E)。最后,奥马珠单抗阻断了花生诱导的花生特异性IgE介导的被动皮肤过敏反应,类似于用来自非花生过敏性供体的血清观察到的结果,该血清没有诱导出染料外渗(图2C)。
总之,实施例1的结果证实了花生过敏的小鼠模型的开发。
实施例2
本实施例证实了AAV载体的设计和表达,该载体包含可操作地连接到编码抗hIgE抗体的核酸序列的启动子。
表达盒由巨细胞病毒(CMV)增强子、与抗hIgE单克隆重链和轻链cDNA序列可操作地连接的鸡β-肌动蛋白启动子(CAG启动子)、以及兔β-球蛋白聚腺苷酸化信号组成。使用人优选的密码子对来自人源化抗IgE抗体SEQ ID NO:10和11的全长重链和轻链氨基酸序列进行回译,并对序列进行优化以改善mRNA稳定性和蛋白质表达。Ig重链和Igκ分泌信号分别被添加到重链和轻链。通过使用弗林蛋白酶切割识别位点(RKRR)下游的Thosea asigna病毒(Tav)2A可切割序列,将重链和轻链克隆到相同的开放阅读框中。两个抗体链以等摩尔比表达自相同的开放阅读框(图3A)。
合成优化的全长抗hIgE cDNA序列并将其克隆到CAG启动子控制下的pAAV质粒中。AAV抗hIgE载体的产生是通过将pAAV质粒连同携带来源于AAV2的载体复制所需的AAV Rep蛋白、AAVrh.10病毒结构(Cap)蛋白VP1、2和3(其定义所产生的AAV载体的血清型);以及E2、E4和VA RNA的腺病毒辅助功能的质粒共转染入人胚胎肾293T细胞(HEK293T;美国典型培养物保藏中心)。通过碘克沙醇梯度和QHP阴离子交换层析纯化AAV抗hIgE载体(称为“AAVrh.10抗hIgE”)。通过定量TaqMan实时PCR分析确定载体基因组滴度。将编码针对尼古丁的无关抗体的载体AAVantiNic(称为“AAVrh.10IgG对照”)用作体内研究的对照。
为了评估在体外AAVrh.10抗hIgE的单克隆抗体的定向表达,用AAVrh.10抗hIgE质粒或编码无关人抗体对照的AAVrh.10IgG对照质粒转染HEK293T细胞,并在72小时后收获上清液。对上清液中抗hIgE抗体表达的评估是通过考马斯亮蓝染色SDS-PAGE和Western分析,使用过氧化物酶缀合的山羊抗人κ轻链抗体和过氧化物酶缀合的山羊抗人IgG抗体和增强的化学发光底物(Bio-Rad,Hercules,CA)。如图3B所示,在细胞培养上清液中检测到抗IgE抗体的重链和轻链。
对于体内研究,6-8周龄的雌性NOD-scidIL2Rγnull(NSG)或雌性Balb/C小鼠通过静脉注射接受单次施用的100μl体积的1011基因组拷贝(gc)的AAVrh.10抗hIgE载体、AAV9抗hIgE载体、AAV8抗hIgE载体、AAVrh.10抗尼古丁载体(对照)或AAVrh.10IgG对照载体。
在0时和各个时间点评估来自尾静脉的血液(100μl),直到第24周。使血样在23℃凝结1小时,然后是30分钟,4℃,然后在1800g下旋转20分钟以收集血清。然后通过ELISA测定抗IgE抗体的浓度。在4℃下用100μl碳酸盐缓冲液(pH9.6)中的0.2μg人IgE涂覆平底96孔EIA/RIA板(Corning,Corning,NY)的孔过夜,然后用0.05%Tween 20(PBS-Tween)的PBS洗涤并用5%奶粉的PBS在23℃封闭60分钟。将连续稀释的血清加入到孔中并在23℃温育60分钟。将板用PBS-Tween洗涤三次,用100μl 1%奶粉的PBS(含1:2000稀释的与辣根过氧化物酶(Abcam,Cambridge,MA)缀合的小鼠抗人IgG)在23℃孵育60分钟。4个洗涤步骤后,将过氧化物酶底物(100μl/孔;Bio-Rad,Hercules,CA)加入到每个孔中,在23℃下孵育15分钟,并加入2%草酸(100μl/孔)停止反应。在415nm下测量吸光度。通过log(OD)-log(稀释)的差值计算抗hIgE抗体的滴度,截断值等于背景吸光度的两倍,并基于采用奥马珠单抗(Genentech,San Francisco,CA)的标准曲线转化为μg/ml,通过PierceTM BCA蛋白分析试剂盒(Life Technologies,Grand Island,NY)进行定量。如图3C中所示,在实验持续期间(44周),人抗IgE表达的水平大于200μg/ml,而对照处理的动物没有检测到人抗IgE。如图3D-3E中所示,对于每种AAV抗IgE载体,证实了人抗IgE的表达,而对照处理的动物没有检测到人抗IgE。
这些数据证实,单次施用AAV-抗hIgE抗体表达盒就能提供高水平、特异性、长期的抗IgE抗体表达。
实施例3
本实施例证实了用AAV-抗hIgE载体预防性治疗以减少对过敏原的过敏反应
为了测试用AAVrh.10抗hIgE预处理是否能保护花生过敏性小鼠,在第-3周用AAVrh.10抗hIgE(1011)或AAVrh.10IgG对照(1011)处理NOD-scid IL2Rγnull小鼠(6-8周龄),然后在第0周用血液单核细胞重建。随后用花生提取物攻击小鼠(图4A)。从载体注射后第2周,每2周评估抗hIgE抗体水平。在第0-4周和第5-8周分别用粗花生提取物致敏和攻击小鼠。在第2、4和8周评估人IgG水平。在第4周时评估人IgE和花生特异性IgE。在第4周时评估游离IgE水平。在每次花生攻击后30分钟评估过敏反应评分,在第6周时评估自发活动,在第7周时评估组胺水平和在第7.5周时评估被动皮肤过敏反应。
在血液单核细胞转移和用花生提取物致敏后,诱导了总人IgE、花生特异性IgE和游离IgE水平(图4B-D)。仅当将特异性过敏原施用至用来自花生过敏性供体的血液单核细胞重建的NSG小鼠时才产生IgE应答。当与用AAVrh.10IgG对照处理的小鼠相比时,用AAVrh.10抗IgE处理的小鼠从第28天起具有显著较低水平的总IgE和花生特异性IgE(p<0.002)。重要的是,与AAVrh.10IgG对照处理的小鼠相比,用AAVrh.10抗hIgE处理的小鼠的游离IgE水平显著更低(p<0.01;图4D)。在用花生提取物灌胃攻击后30分钟评价过敏反应。引人注目的是,与接受对照载体的那些相比,AAVrh.10抗hIgE处理的小鼠表现出如临床表型所定义的严重程度低的多的过敏表型、抑制的移动、较低的过敏反应评分、降低的组胺释放和降低的PCA(图5A-D)。
这些数据证实了AAVrh.10抗hIgE载体的单一预防性治疗就能减少或抑制免疫应答或过敏反应。
实施例4
本实施例证实了暴露于抗原后载体施用的有效性。
为了确定在小鼠已经用花生提取物致敏并且表现出花生提取物诱导的过敏反应之后,用AAVrh.10抗hIgE处理是否可以保护花生过敏性小鼠,在第0周时用血液单核细胞重建NOD-scidIL2Rγnull小鼠,然后在第0-4周和第5-10周时分别用粗花生提取物致敏和攻击。在与花生攻击(第5周)相关的过敏反应的第一迹象之后,向小鼠施用AAVrh.10抗hIgE、AAVrh.10IgG对照或200μl 0.9%NaCl中的250μg人源化抗IgE mAb奥马珠单抗。以固定间隔(每2周)评估抗hIgE抗体水平(图6A)。在第2、4和8周时评估人IgG水平。在第4和第6周时评估人IgE,在第4和第6周时评估游离IgE水平,并在第4周时评估花生特异性IgE。在第7周和第10周的每次花生攻击后30分钟评估过敏反应评分和临床评分。在第7周和第10周时评估自发活动,在第6周和第9周时评估组胺水平,并在第7周和第10周时评估被动皮肤过敏反应。
在血液单核细胞转移并用花生提取物致敏之后,仅当将特异性过敏原注射到用来自花生过敏性供体的血液单核细胞重建的NSG小鼠组中时才产生IgE应答,并在花生刺激后持续(图6B)。在治疗前1周,在第4周时完成花生致敏后,所有小鼠都产生了花生特异性IgE(图6C)。与AAVrh.10IgG对照处理的小鼠相比,AAVrh.10抗hIgE处理的小鼠的游离IgE水平在治疗后1周显著降低(第6周;p<0.01)(图6D)。在第10周时,用AAVrh.10抗hIgE治疗5周后,花生过敏性小鼠不具有临床症状,而用奥马珠单抗处理的花生过敏性小鼠具有眼和口鼻周围浮肿、毛发直立、皮毛瘙痒和皱褶(图7A)。在第7周时,治疗2周后,AAVrh.10抗hIgE和奥马珠单抗治疗的小鼠均比对照显著增加移动性(图7B,左),但在第10周时,治疗5周后,AAVrh.10抗hIgE小鼠比奥马珠单抗处理的小鼠显著增加移动性(图7B右;到第10周时,所有对照小鼠已经死亡)。与临床表型和移动数据相一致,与对照相比,AAVrh.10抗hIgE和奥马珠单抗处理的小鼠在第7周(治疗后2周)时的过敏反应评分显著更低,但在第10周(治疗后5周)时仅有AAVrh.10抗hIgE疗法继续有效,在第7周时奥马珠单抗小鼠与对照小鼠相似(到第10周时所有对照小鼠已经死亡;图7C)。对于血浆组胺水平(图7D)和被动皮肤过敏反应(图7E)具有相同的观察结果。
最引人注目的观察结果是生存分析。治疗后40天,仅有接受AAVrh.10抗hIgE的小鼠被保护免于死亡(图8)。90%的AAVrh.10抗hIgE小鼠(9/10)在治疗后存活直至40天(最后一个评估时间点),而70%(7/10)的奥马珠单抗处理的小鼠死亡且89%(8/9)AAVrh.10IgG对照小鼠死亡。与小鼠中的异种移植物抗宿主病(GVHD)相同,炎症和免疫浸润均出现在用来自过敏性和非过敏供体的单核细胞重建后第6到第7周的人源化小鼠的肺和小肠中(表I)。在肺组织中见到了肺部GVHD的诊断特征,即淋巴组织细胞、浆细胞和中性粒细胞血管周炎和细支气管周炎、弥漫性间质中性粒细胞增多和多灶性细支气管上皮内嗜酸性包涵体。小肠组织显示出符合GVHD的组织学特征,即嗜酸性粒细胞、中性粒细胞和淋巴细胞浸润。
本文引用的所有参考文献,包括出版物、专利申请和专利,在此通过引用并入,其程度如同每个参考文献单独且具体地指示为通过引用并入并以其全部内容在本文中阐述。
除非在本文中另有指示或明显与上下文矛盾,在描述本发明的上下文中(特别是在以下权利要求的上下文中)使用的术语“一”、“一个”、“该”和“至少一个”以及类似的指示物应被解释为涵盖单数和复数两种情况。除非在本文中另有指示或明显与上下文矛盾,使用的术语“至少一个”后接一个或多个项目的列表(例如,“A和B中的至少一个”)应理解为是指从列出的项目(A或B)或所列项目(A和B)中的两个或更多个的任何组合。除非另有说明,术语“包含”、“具有”、“包括”和“含有”应被解释为开放式术语(即是指“包括但不限于”)。除非在本文中另有指示,本文中数值范围的记载仅旨在用作单独提及落在该范围内的每个单独值的速记方法,并且将每个单独的值并入本说明书中,如同其在本文中单独列举一样。除非在本文中另有指示或在其它方面明显与上下文矛盾,本文描述的所有方法可以以任何合适的顺序进行。除非另有要求,本文提供的任何和所有实例或示例性语言(例如“例如”)的使用仅旨在更好地阐明本发明,并且不对本发明的范围构成限制。说明书中的任何语言不应被解释为指示任何未被要求保护的要素对于本发明的实践是必需的。
这里描述了本发明的优选实施例,包括发明人已知的用于实施本发明的最佳模式。在阅读前面的描述之后,那些优选实施例的变化对于本领域的普通技术人员来说会变得明显。本发明人期望本领域技术人员适当地采用这样的变化,并且本发明人希望以不同于在此具体描述的方式来实践本发明。因此,本发明包括适用法律允许的所附权利要求书中记载的主题的所有修改和等同物。此外,除非本文另有说明或者与上下文明显矛盾,否则本发明涵盖上述要素在其所有可能变型中的任何组合。
序列表
<110> 康奈尔大学
<120> 基因治疗以预防对过敏原的反应
<130> 723279
<150> US 62/314,740
<151> 2016-03-29
<150> US 62/145,035
<151> 2015-04-09
<160> 11
<170> PatentIn version 3.5
<210> 1
<211> 36
<212> DNA
<213> 人工序列
<220>
<223> 合成重链 CDR-H3
<400> 1
cgccgtgagc ggctacagca tcaccagcgg ctacag 36
<210> 2
<211> 30
<212> DNA
<213> 人工序列
<220>
<223> 合成重链 CDR-H2
<400> 2
cagcatcacc tacgacggca gcaccaacta 30
<210> 3
<211> 42
<212> DNA
<213> 人工序列
<220>
<223> 合成重链 CDR-H3
<400> 3
cgctcggggc agccactact tcggccactg gcacttcgcc gt 42
<210> 4
<211> 45
<212> DNA
<213> 人工序列
<220>
<223> 合成轻链 CDR-L1
<400> 4
ccgggctagc cagagcgtgg actacgacgg cgacagctac atgaa 45
<210> 5
<211> 24
<212> DNA
<213> 人工序列
<220>
<223> 合成轻链 CDR-L2
<400> 5
ctacgctgcc agctacctgg agag 24
<210> 6
<211> 27
<212> DNA
<213> 人工序列
<220>
<223> 合成轻链 CDR-L3
<400> 6
ccagcagagc cacgaggacc cctacac 27
<210> 7
<211> 363
<212> DNA
<213> 人工序列
<220>
<223> 合成重链可变区
<400> 7
gaggtgcagc tggtggagag cggcggtggc ctggtgcagc ctggtggtag cctgcgcctg 60
agctgcgccg tgagcggcta cagcatcacc agcggctaca gctggaactg gatccggcag 120
gcccctggca agggcctgga gtgggtggcc agcatcacct acgacggcag caccaactac 180
gccgacagcg tgaagggccg gttcaccatc agccgggacg acagcaagaa caccttctac 240
ctgcagatga actccctgcg ggccgaggac accgccgtgt actactgcgc tcggggcagc 300
cactacttcg gccactggca cttcgccgtg tggggtcagg gcaccctggt gaccgtgagc 360
agc 363
<210> 8
<211> 330
<212> DNA
<213> 人工序列
<220>
<223> 合成轻链可变区
<400> 8
gacatccagc tgacccagag ccctagcagc ctgagcgcca gcgtgggcga ccgggtgacc 60
atcacctgcc gggctagcca gagcgtggac tacgacggcg acagctacat gaattggtac 120
cagcagaagc ccggcaaggc tcccaagctg ctgatctacg ctgccagcta cctggagagc 180
ggcgtgccca gccggttcag cggcagcggc agcggcaccg acttcaccct gaccatcagc 240
agcctgcagc ccgaggactt cgccacctac tactgccagc agagccacga ggacccctac 300
accttcggcc agggcaccaa ggtggagatc 330
<210> 9
<211> 2214
<212> DNA
<213> 人工序列
<220>
<223> 合成抗IgE抗体
<400> 9
atggagttcg gcctgagctg gctgttcctg gtggccatcc tgaagggcgt gcagtgcgag 60
gtgcagctgg tggagagcgg cggtggcctg gtgcagcctg gtggtagcct gcgcctgagc 120
tgcgccgtga gcggctacag catcaccagc ggctacagct ggaactggat ccggcaggcc 180
cctggcaagg gcctggagtg ggtggccagc atcacctacg acggcagcac caactacgcc 240
gacagcgtga agggccggtt caccatcagc cgggacgaca gcaagaacac cttctacctg 300
cagatgaact ccctgcgggc cgaggacacc gccgtgtact actgcgctcg gggcagccac 360
tacttcggcc actggcactt cgccgtgtgg ggtcagggca ccctggtgac cgtgagcagc 420
gccagcacca agggccccag cgtgttccct ctggctccct cttccaaatc caccagcggc 480
ggcaccgctg ccctgggctg cctggtgaag gactacttcc ccgagcccgt gaccgtgagc 540
tggaacagcg gcgccctgac cagcggcgtg cacaccttcc ctgccgtgct gcaatccagc 600
ggcctgtaca gcctgagcag cgtcgtgacc gtgcccagca gcagcctggg cacccagacc 660
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggccgagccc 720
aagagctgcg acaagaccca cacctgccct ccctgccccg cccctgagct gctcggcgga 780
cccagcgtgt tcctgttccc tcccaagccc aaggacaccc tgatgatcag ccggacccct 840
gaggtgacct gcgtggtcgt ggacgtgagc cacgaggacc ccgaggtgaa gttcaactgg 900
tatgtggacg gcgtggaggt gcacaacgcc aagaccaagc cccgggagga gcagtacaac 960
agcacctacc gggtggtgag cgtgctgacc gtgctgcacc aggactggct gaacggcaag 1020
gagtacaagt gcaaggtctc caacaaggcc ctgcctgccc ctatcgagaa gaccatcagc 1080
aaggccaagg gccagccccg ggagccccag gtgtacaccc tgcctcctag ccgggacgag 1140
ctgaccaaga accaggtctc cctgacctgc ctggtgaagg gcttctaccc cagcgacatc 1200
gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac ccctcccgtg 1260
ctggacagcg acggcagctt cttcctgtac tccaagctga ccgtggacaa gagccggtgg 1320
cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1380
cagaagagcc tgagcctgag ccccggacgg aagcggagga gcggcgctcc tgtgaagcag 1440
accctgaact tcgacctgct gaagctggcc ggcgacgtgg agagcaaccc tgggcctatg 1500
aagtacctgc tgcccaccgc cgctgctggc ctgctcctgc tcgctgccca acccgctgct 1560
gacatccagc tgacccagag ccctagcagc ctgagcgcca gcgtgggcga ccgggtgacc 1620
atcacctgcc gggctagcca gagcgtggac tacgacggcg acagctacat gaattggtac 1680
cagcagaagc ccggcaaggc tcccaagctg ctgatctacg ctgccagcta cctggagagc 1740
ggcgtgccca gccggttcag cggcagcggc agcggcaccg acttcaccct gaccatcagc 1800
agcctgcagc ccgaggactt cgccacctac tactgccagc agagccacga ggacccctac 1860
accttcggcc agggcaccaa ggtggagatc aagcggaccg tggccgctcc tagcgtgttc 1920
atcttccctc cctccgacga gcagctgaag agcggcaccg ccagcgtggt gtgcctgctg 1980
aacaacttct accctcggga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 2040
ggcaacagcc aggagagcgt gaccgagcag gacagcaagg acagcaccta cagcctgagc 2100
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 2160
acccaccagg gcctgagcag ccccgtgacc aagagcttca accggggcga gtgc 2214
<210> 10
<211> 1407
<212> DNA
<213> 人工序列
<220>
<223> 合成奥马珠单抗重链
<400> 10
atggagttcg gcctgagctg gctgttcctg gtggccatcc tgaagggcgt gcagtgcgag 60
gtgcagctgg tggagagcgg cggtggcctg gtgcagcctg gtggtagcct gcgcctgagc 120
tgcgccgtga gcggctacag catcaccagc ggctacagct ggaactggat ccggcaggcc 180
cctggcaagg gcctggagtg ggtggccagc atcacctacg acggcagcac caactacgcc 240
gacagcgtga agggccggtt caccatcagc cgggacgaca gcaagaacac cttctacctg 300
cagatgaact ccctgcgggc cgaggacacc gccgtgtact actgcgctcg gggcagccac 360
tacttcggcc actggcactt cgccgtgtgg ggtcagggca ccctggtgac cgtgagcagc 420
gccagcacca agggccccag cgtgttccct ctggctccct cttccaaatc caccagcggc 480
ggcaccgctg ccctgggctg cctggtgaag gactacttcc ccgagcccgt gaccgtgagc 540
tggaacagcg gcgccctgac cagcggcgtg cacaccttcc ctgccgtgct gcaatccagc 600
ggcctgtaca gcctgagcag cgtcgtgacc gtgcccagca gcagcctggg cacccagacc 660
tacatctgca acgtgaacca caagcccagc aacaccaagg tggacaagaa ggccgagccc 720
aagagctgcg acaagaccca cacctgccct ccctgccccg cccctgagct gctcggcgga 780
cccagcgtgt tcctgttccc tcccaagccc aaggacaccc tgatgatcag ccggacccct 840
gaggtgacct gcgtggtcgt ggacgtgagc cacgaggacc ccgaggtgaa gttcaactgg 900
tatgtggacg gcgtggaggt gcacaacgcc aagaccaagc cccgggagga gcagtacaac 960
agcacctacc gggtggtgag cgtgctgacc gtgctgcacc aggactggct gaacggcaag 1020
gagtacaagt gcaaggtctc caacaaggcc ctgcctgccc ctatcgagaa gaccatcagc 1080
aaggccaagg gccagccccg ggagccccag gtgtacaccc tgcctcctag ccgggacgag 1140
ctgaccaaga accaggtctc cctgacctgc ctggtgaagg gcttctaccc cagcgacatc 1200
gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac ccctcccgtg 1260
ctggacagcg acggcagctt cttcctgtac tccaagctga ccgtggacaa gagccggtgg 1320
cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa ccactacacc 1380
cagaagagcc tgagcctgag ccccgga 1407
<210> 11
<211> 645
<212> DNA
<213> 人工序列
<220>
<223> 合成奥马珠单抗轻链
<400> 11
gacatccagc tgacccagag ccctagcagc ctgagcgcca gcgtgggcga ccgggtgacc 60
atcacctgcc gggctagcca gagcgtggac tacgacggcg acagctacat gaattggtac 120
cagcagaagc ccggcaaggc tcccaagctg ctgatctacg ctgccagcta cctggagagc 180
ggcgtgccca gccggttcag cggcagcggc agcggcaccg acttcaccct gaccatcagc 240
agcctgcagc ccgaggactt cgccacctac tactgccagc agagccacga ggacccctac 300
accttcggcc agggcaccaa ggtggagatc aagcggaccg tggccgctcc tagcgtgttc 360
atcttccctc cctccgacga gcagctgaag agcggcaccg ccagcgtggt gtgcctgctg 420
aacaacttct accctcggga ggccaaggtg cagtggaagg tggacaacgc cctgcagagc 480
ggcaacagcc aggagagcgt gaccgagcag gacagcaagg acagcaccta cagcctgagc 540
agcaccctga ccctgagcaa ggccgactac gagaagcaca aggtgtacgc ctgcgaggtg 600
acccaccagg gcctgagcag ccccgtgacc aagagcttca accgg 645
Claims (20)
1.一种用于减少哺乳动物中对过敏原的免疫应答或过敏反应的腺相关病毒(AAV)载体,其包含可操作地连接到核酸序列的启动子,
其中所述核酸序列编码抗IgE抗体或其抗原结合片段,其中所述抗体或其抗原结合片段包含重链多肽以及轻链多肽,
其中所述重链多肽包含三个互补决定区(CDR),其中CDR-H1由SEQ ID NO:1的核酸序列编码,CDR-H2由SEQ ID NO:2的核酸序列编码,CDR-H3由SEQ ID NO:3的核酸序列编码;
其中所述轻链多肽包含三个CDR,其中CDR-L1由SEQ ID NO:4的核酸序列编码,CDR-L2由SEQ ID NO:5的核酸序列编码,CDR-L3由SEQ ID NO:6的核酸序列编码;和
其中所述AAV载体是腺相关病毒血清型rh.10载体。
2.权利要求1所述的AAV载体,其中所述启动子是组成型活性启动子。
3.权利要求1所述的AAV载体,其中所述启动子是细胞类型特异性启动子。
4.权利要求1所述的AAV载体,其中所述启动子是诱导型启动子。
5.权利要求4所述的AAV载体,其中所述组成型活性启动子是鸡β-肌动蛋白启动子。
6.权利要求1所述的AAV载体,其中所述抗IgE抗体包含由SEQ ID NO:7的核酸序列编码的重链可变区和由SEQ ID NO:8的核酸序列编码的轻链可变区。
7.权利要求1所述的AAV载体,其中所述AAV载体包含SEQ ID NO:9的核酸序列。
8.权利要求1所述的AAV载体,其中所述AAV载体包含编码奥马珠单抗的核酸序列。
9.权利要求1所述的AAV载体,其中所述核酸序列编码抗IgE抗体片段,所述抗IgE抗体片段选自由F(ab’)2、Fab'、Fab、Fv、scFv、dsFv、dAb和单链结合多肽组成的组。
10.一种组合物,其包含权利要求1-9中任一项所述的AAV载体和药学上可接受的运载体。
11.权利要求1-9中任一项所述的AAV载体在制备用于减少哺乳动物中对过敏原的免疫应答或过敏反应的药物中的用途,所述药物用于向所述哺乳动物施用,由此表达所述核酸,并且减少针对所述过敏原的免疫应答或过敏反应。
12.权利要求11所述的用途,其中所述药物在30天内施用至所述哺乳动物不超过一次。
13.权利要求11所述的用途,其中在施用所述药物后,所述哺乳动物表达治疗或预防水平的抗IgE抗体或其抗原结合片段,30天或更长时间。
14.权利要求11所述的用途,其中所述药物预防性地施用于所述哺乳动物。
15.权利要求11所述的用途,其中所述过敏原是食物过敏原。
16.权利要求11所述的用途,其中所述过敏反应是过敏性休克。
17.权利要求11所述的用途,其中所述哺乳动物是人。
18.权利要求11所述的用途,其中所述药物通过选自由口内、肌内、透皮、静脉内、动脉内、皮下、皮内和腹膜内组成的组的给药途径施用于所述哺乳动物。
19.权利要求11所述的用途,其中所述过敏原选自花粉、尘螨、昆虫毒液、花生和树坚果。
20.权利要求11所述的用途,其中所述过敏原是蜂螯毒液。
Applications Claiming Priority (5)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
US201562145035P | 2015-04-09 | 2015-04-09 | |
US62/145,035 | 2015-04-09 | ||
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WO2017189964A2 (en) | 2016-04-29 | 2017-11-02 | Voyager Therapeutics, Inc. | Compositions for the treatment of disease |
EP3448874A4 (en) | 2016-04-29 | 2020-04-22 | Voyager Therapeutics, Inc. | COMPOSITIONS FOR TREATING A DISEASE |
WO2018139634A1 (ja) * | 2017-01-30 | 2018-08-02 | 学校法人日本医科大学 | アデノ随伴ウイルス(aav)キャプシドタンパク質の変異体 |
CN112041338A (zh) * | 2017-12-29 | 2020-12-04 | 康奈尔大学 | 嗜酸性粒细胞病症的基因疗法 |
CN110066768A (zh) * | 2018-08-24 | 2019-07-30 | 中国疾病预防控制中心营养与健康所 | 表达hFcεRIα的RBL-2H3细胞株的构建方法 |
CN111351921A (zh) * | 2018-12-21 | 2020-06-30 | 泰州迈博太科药业有限公司 | 食物致敏动物模型、构建方法及其应用 |
CN112023062A (zh) * | 2020-09-18 | 2020-12-04 | 北京基因安科技有限公司 | 一个用可溶性IgE受体抑制过敏反应的方法 |
WO2022109093A1 (en) * | 2020-11-18 | 2022-05-27 | IgGenix, Inc. | Compositions and methods for treating and suppressing allergic responses |
Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999001556A2 (en) * | 1997-07-02 | 1999-01-14 | Genentech, Inc. | IMPROVED ANTI-IgE ANTIBODIES AND METHOD OF IMPROVING POLYPEPTIDES |
US20100143262A1 (en) * | 2008-10-17 | 2010-06-10 | Rudolph Valenta | Method for Identifying Allergenic Proteins And Peptides |
US20110064726A1 (en) * | 2007-04-02 | 2011-03-17 | Pfizer Inc. | ANTI IgE ANTIBODIES |
Family Cites Families (25)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US4797368A (en) | 1985-03-15 | 1989-01-10 | The United States Of America As Represented By The Department Of Health And Human Services | Adeno-associated virus as eukaryotic expression vector |
US4863457A (en) | 1986-11-24 | 1989-09-05 | Lee David A | Drug delivery device |
US5378475A (en) | 1991-02-21 | 1995-01-03 | University Of Kentucky Research Foundation | Sustained release drug delivery devices |
US5464758A (en) | 1993-06-14 | 1995-11-07 | Gossen; Manfred | Tight control of gene expression in eucaryotic cells by tetracycline-responsive promoters |
US5814618A (en) | 1993-06-14 | 1998-09-29 | Basf Aktiengesellschaft | Methods for regulating gene expression |
US5443505A (en) | 1993-11-15 | 1995-08-22 | Oculex Pharmaceuticals, Inc. | Biocompatible ocular implants |
US6342390B1 (en) | 1994-11-23 | 2002-01-29 | The United States Of America As Represented By The Secretary Of Health And Human Services | Lipid vesicles containing adeno-associated virus rep protein for transgene integration and gene therapy |
CN1131314C (zh) * | 1996-04-29 | 2003-12-17 | 仁文有限公司 | 含有过敏原基因的重组真核载体及其应用 |
EP2361635A3 (en) * | 2000-08-30 | 2011-09-14 | Pfizer Products Inc. | Anti IgE vaccines |
FR2814642B1 (fr) | 2000-10-03 | 2005-07-01 | Ass Pour Le Dev De La Rech En | Souris transgenique pour la recombinaison ciblee mediee par la cre-er modifiee |
US6723551B2 (en) | 2001-11-09 | 2004-04-20 | The United States Of America As Represented By The Department Of Health And Human Services | Production of adeno-associated virus in insect cells |
EP1585768A2 (en) * | 2003-01-23 | 2005-10-19 | Genentech, Inc. | Methods for producing humanized antibodies and improving yield of antibodies or antigen binding fragments in cell culture |
CN1317304C (zh) * | 2004-03-10 | 2007-05-23 | 伍祥贵 | 一种融合蛋白及其编码基因与表达方法和应用 |
WO2006017325A2 (en) * | 2004-07-13 | 2006-02-16 | Cell Genesys, Inc. | Aav vector compositions and methods for enhanced expression of immunoglobulins the same |
CN101062949B (zh) * | 2006-04-26 | 2011-07-27 | 上海抗体药物国家工程研究中心有限公司 | 重组抗人IgE单克隆抗体及其制备方法和用途 |
DE102007001370A1 (de) * | 2007-01-09 | 2008-07-10 | Curevac Gmbh | RNA-kodierte Antikörper |
US20100136030A1 (en) | 2007-02-27 | 2010-06-03 | Lamhamedi-Cherradi Salah-Eddine | Antagonist ox40 antibodies and their use in the treatment of inflammatory and autoimmune diseases |
US8242093B2 (en) * | 2008-02-07 | 2012-08-14 | Ceregene, Inc. | Rescue of photoreceptors by intravitreal administration of an expression vector encoding a therapeutic protein |
JP5634272B2 (ja) * | 2008-03-14 | 2014-12-03 | ヒューマンザイム リミテッド | ヒト細胞発現系を用いる真正ヒトタンパク質の組換え生産 |
GB0917044D0 (en) | 2009-09-29 | 2009-11-18 | Cytoguide As | Agents, uses and methods |
MX2013005847A (es) | 2010-11-24 | 2013-12-12 | Glaxo Group Ltd | Proteinas de union a antigeno multiespecificas que eligen como blanco a hgf. |
MX356947B (es) * | 2011-08-23 | 2018-06-20 | Roche Glycart Ag | Anticuerpos bioespecíficos específicos para antígenos que activan células t y un antígeno tumoral y métodos de uso. |
EP3539568A1 (en) * | 2011-11-22 | 2019-09-18 | The Children's Hospital of Philadelphia | Virus vectors for highly efficient transgene delivery |
CN105452458A (zh) * | 2013-06-07 | 2016-03-30 | 加利福尼亚大学董事会 | 用于治疗心力衰竭和增加心脏功能的不能产生环腺苷酸的腺苷酸环化酶和组合物和方法 |
US11408899B2 (en) * | 2013-07-12 | 2022-08-09 | The Children's Hospital Of Philadelphia | AAV vector and assay for anti-AAV (adeno-associated virus) neutralizing antibodies |
-
2016
- 2016-04-11 WO PCT/US2016/026977 patent/WO2016164920A1/en unknown
- 2016-04-11 EP EP16777501.4A patent/EP3280445B1/en active Active
- 2016-04-11 JP JP2017553166A patent/JP6878301B2/ja active Active
- 2016-04-11 CA CA2982213A patent/CA2982213C/en active Active
- 2016-04-11 US US15/096,120 patent/US10293059B2/en active Active
- 2016-04-11 CN CN201680033587.3A patent/CN107635584B/zh active Active
-
2021
- 2021-02-15 JP JP2021022067A patent/JP2021097677A/ja active Pending
Patent Citations (3)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO1999001556A2 (en) * | 1997-07-02 | 1999-01-14 | Genentech, Inc. | IMPROVED ANTI-IgE ANTIBODIES AND METHOD OF IMPROVING POLYPEPTIDES |
US20110064726A1 (en) * | 2007-04-02 | 2011-03-17 | Pfizer Inc. | ANTI IgE ANTIBODIES |
US20100143262A1 (en) * | 2008-10-17 | 2010-06-10 | Rudolph Valenta | Method for Identifying Allergenic Proteins And Peptides |
Non-Patent Citations (1)
Title |
---|
Bicistronic expression plasmid encoding allergen and anti-IgE single chain variable fragment antibody as a novel DNA vaccine for allergy therapy and prevention;Nariman Aghaei Bandbon Balenga等;《Medical Hypotheses》;20060302;第67卷(第1期);摘要 * |
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