CN115137814A - 一种肿瘤疫苗佐剂 - Google Patents
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Abstract
本发明公开了一种肿瘤疫苗佐剂,本发明所需的肿瘤疫苗佐剂的主要成分是GM‑CSF、白细胞介素‑2和IFN‑γ,本发明所述新型疫苗佐剂为免疫刺激型佐剂,具有免疫调节活性作用,在体内诱导T好型免疫应答,壳作为治疗与预防肿瘤用的疫苗佐剂。
Description
技术领域
本发明涉及疫苗佐剂相关领域,具体是一种肿瘤疫苗佐剂。
背景技术
佐剂是一种非特异性免疫增强物质,当与抗原一起注射或预先注入机体时,可增强机体对抗原的免疫应答或改变免疫应答类型,目前常用疫苗佐剂有氢氧化铝佐剂、短小棒状杆菌、脂多糖、细胞因子、明矾等,但是以上这些佐剂对细胞免疫作用不明显,尤其是对以细胞免疫为主的疫苗,如对高纯化的第二代重组疫苗和第三代DNA疫苗,效果甚微,现有以上这些佐剂在临床上的应用已经满足不了现代疫苗技术发展的需求。
恶性肿瘤是严重威胁人类的健康的多发病和常见病,手术、化疗和放疗是治疗恶性肿瘤的三大传统疗法,肿瘤的免疫治疗近年来发展迅速,提高肿瘤的免疫原性,制备高效的肿瘤疫苗已成为免疫治疗的热点,最有效的疫苗必须既能激活CD4+T细胞又能激活CD8+T细胞,因此基于树突状细胞的免疫治疗和肿瘤疫苗具有极大的开发及临床应用价值。
发明内容
因此,为了解决上述不足,本发明在此提供一种肿瘤疫苗佐剂。
本发明是这样实现的,构造一种肿瘤疫苗佐剂,主要成分是GM-CSF、白细胞介素-2和IFN-γ。
优选的,其中所述的GM-CSF的质量百分比为15%,GM-CSF是由胺基酸所組成。
优选的,其中所述的白细胞介素-2的质量百分比为40%-70%,白细胞介素 -2是由胺基酸所組成。
优选的,其中所述的IFN-γ的质量百分比为15%-35%。
优选的,其中所述的IFN-γ是由胺基酸所組成。
优选的,该佐剂在制备治疗和预防用疫苗佐剂中的应用。
优选的,其抗原和佐剂比例为:1∶1-10∶1-2.5。
优选的,所述抗原为γ干扰素、白细胞介素-2、粒细胞-巨噬细胞集落刺激因子。
优选的,该佐剂包括通过化学合成方式来合成肽的过程及其制备方法。
附图说明
图1是免疫细胞在实验生物内的分裂量示意图;
图2是五只实验鼠的注射疫苗的免疫率图。
具体实施方式
下面将结合附图1-2对本发明进行详细说明,对本发明实施例中的技术方案进行清楚、完整地描述,显然,所描述的实施例仅仅是本发明一部分实施例,而不是全部的实施例。基于本发明中的实施例,本领域普通技术人员在没有做出创造性劳动前提下所获得的所有其他实施例,都属于本发明保护的范围。
本文所述的术语“抗肿瘤疫苗佐剂”,也称作“疫苗佐剂”,是指能够非特异性的增强免疫的物质,优选多肽物质,其能够将体液免疫转变为细胞免疫,改变免疫球蛋白的种类和亚类,消除免疫耐受性从而增强免疫反应。其特征在于:能促进体液和细胞介导的免疫,也能作用于弱免疫性抗原,而且不引起有害的副作用,能以不同途径免疫,也能用于不同抗原,能在免疫抑制个体中发挥作用。
所述两个以上的基因可为相同基因或不同基因,本发明中优选为不同基因,更优选为来源于不同生物物种的基因,例如,两个以上的基因可为来源于微生物与高等生物的基因。
本文所述的“氨基酸序列”是指两个以上的氨基酸通过共价键结合的化合物。其中氨基酸可为天然氨基酸或氨基酸衍生物,或者人工修改的氨基酸。
本文所述的融合蛋白可通过本领域内已知的任何手段来获得,例如通过 DNA重组技术,用不同基因融合而成的基因在生物表达系统中表达而产生。另外,也可通过化学合成法合成不同的氨基酸序列,最后通过连接不同氨基酸序列而成,或者通过化学合成法延长氨基酸链而成。
将细胞在37℃湿度培养箱中孵育直到细胞出现病毒感染迹象,将感染的细胞反复冻融,使细胞裂解,由于鞭毛蛋白与其配体TLR5结合,将激活下游NF- κB、MEKK-p38MAPK等信号通路,我们将Flagp96在DC中共培养,在不同时间点收获细胞,提取蛋白,免疫迹象试验方法检测MAPK及其磷酸化,结果表明,这些Flagp96能够激活DC中的MAPK及其磷酸化。
幼稚PmelCD8+T细胞(2×106)通过尾静脉注射给C57BL/6小鼠(每组5只),用1×106负载gp96、Flagp96与黑色素瘤抗原肽gp10025-33复合物、OVA的 DC及负载gp10025-33的Flagellin-DC尾静脉注射免疫小鼠,不同的时间点,收集脾、回流淋巴结,流式分析检测Pmel特异CD8+T细胞的增殖;在高尔基体阻断剂存在的情况下,gp10025-33肽刺激4小时后,细胞内染色分析CD8+T 细胞的IL-2、IFN-γ分泌,明显提高DNA疫苗抗肿瘤免疫作用。
记录了实施组与对比组小鼠的肿瘤生长变化,通过试验,实施组的小鼠在免疫后第135天依然存活,且身体看不到肿瘤块;而对比组的小鼠在第35天肿瘤扩散严重。另外,与未包裹的mcGMCSF-293细胞系(剂量:1×106细胞))免疫组相比,包裹的mcGMCSF-293细胞系(剂量:1×105细胞))免疫组仅需要较少的免疫佐剂便可达到完全的保护,
以上显示和描述了本发明的基本原理和主要特征和本发明的优点,并且本发明使用到的标准零件均可以从市场上购买,异形件根据说明书的和附图的记载均可以进行订制,各个零件的具体连接方式均采用现有技术中成熟的螺栓铆钉、焊接等常规手段,机械、零件和设备均采用现有技术中,常规的型号,加上电路连接采用现有技术中常规的连接方式,在此不再详述。
对所公开的实施例的上述说明,使本领域专业技术人员能够实现或使用本发明。对这些实施例的多种修改对本领域的专业技术人员来说将是显而易见的,本文中所定义的一般原理可以在不脱离本发明的精神或范围的情况下,在其它实施例中实现。因此,本发明将不会被限制于本文所示的这些实施例,而是要符合与本文所公开的原理和新颖特点相一致的最宽的范围。
Claims (9)
1.一种肿瘤疫苗佐剂,其主要成分是GM-CSF、白细胞介素-2和IFN-γ。
2.根据权利要求1所述的一种肿瘤疫苗佐剂,其中所述的GM-CSF的质量百分比为15%,GM-CSF是由胺基酸所組成。
3.根据权利要求1所述的一种肿瘤疫苗佐剂,其中所述的白细胞介素-2的质量百分比为40%-70%,白细胞介素-2是由胺基酸所組成。
4.根据权利要求1所述的一种肿瘤疫苗佐剂,其中所述的IFN-γ的质量百分比为15%-35%。
5.根据权利要求1所述的一种肿瘤疫苗佐剂,其中所述的IFN-γ是由胺基酸所組成。
6.根据权利要求1所述的一种肿瘤疫苗佐剂,该佐剂在制备治疗和预防用疫苗佐剂中的应用。
7.根据权利要求1所述的一种肿瘤疫苗佐剂,其抗原和佐剂比例为:1∶1-10∶1-2.5。
8.根据权利要求6所述一种肿瘤疫苗佐剂,其特征在于:所述抗原为γ干扰素、白细胞介素-2、粒细胞-巨噬细胞集落刺激因子。
9.根据权利要求1所述的一种肿瘤疫苗佐剂,该佐剂包括通过化学合成方式来合成肽的过程及其制备方法。
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CN105473157A (zh) * | 2013-08-21 | 2016-04-06 | 库瑞瓦格股份公司 | 组合疫苗 |
CN106636138A (zh) * | 2017-01-12 | 2017-05-10 | 李增魁 | 一种针对羊种3型布鲁氏菌感染的dna疫苗及其制备方法 |
CN107157933A (zh) * | 2017-05-04 | 2017-09-15 | 同济大学 | 一种蛋白自组装新型纳米疫苗及其制备方法 |
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