Anti-Tumor Effificacy of an Adjuvant Built-In Nanovaccine Based on Ubiquitinated Proteins from Tumor Cell
Anti-Tumor Effificacy of an Adjuvant Built-In Nanovaccine Based on Ubiquitinated Proteins from Tumor Cell
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基于肿瘤细胞泛素化蛋白的佐剂内置纳米疫苗的抗肿瘤功效
DOI：
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发表时间：
2020
影响因子：
8
通讯作者：
Ning Pan
中科院分区：
文献类型：
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作者：
Fang Huang;Jinjin Zhao;Yiting Wei;Zhifa Wen;Yue Zhang;Xuru Wang;Yanfei Shen;Li-xin Wang;Ning Pan
背景与目的：我们之前已从肿瘤细胞裂解物中鉴定出泛素化蛋白（UPs），在不同的小鼠肿瘤模型中，其作为一种有前景的癌症免疫治疗疫苗。在本研究中，我们旨在通过一种简单方法开发一种高效的内置治疗佐剂纳米疫苗（α -Al₂O₃ -UPs），利用与Vx3蛋白共价偶联的α -Al₂O₃纳米颗粒（α -Al₂O₃ -CONH -Vx3），可高效便捷地富集肿瘤细胞中的UPs。
方法：用4 -羟基苯甲酸对α -Al₂O₃纳米颗粒进行修饰，随后与泛素结合蛋白Vx3偶联。接着用其从4T1细胞裂解物中富集UPs。检测α -Al₂O₃ -CONH -Vx3富集UPs的稳定性和效率。随后在体外检测α -Al₂O₃ -UPs激活树突状细胞（DCs）的能力。用灭活的肿瘤细胞重新刺激接种疫苗小鼠的脾细胞，通过酶联免疫吸附试验（ELISA）和流式细胞术检测γ -干扰素（IFN -γ）的分泌情况。此外，在4T1荷瘤小鼠中检测α -Al₂O₃ -UPs单独使用及与化疗联合使用的治疗效果。
结果：我们的结果表明，成功合成了α -Al₂O₃ -UPs，并且通过新方法富集到了肿瘤细胞裂解物中大量的UPs。体外研究显示，与α -Al₂O₃纳米颗粒和UPs的物理混合物（α -Al₂O₃ +UPs）相比，α -Al₂O₃ -UPs刺激导致DCs上CD80、CD86、I类主要组织相容性复合体（MHC class I）和II类主要组织相容性复合体（MHC class II）的上调幅度更大，表明其激活DCs的能力更强。此外，α -Al₂O₃ -UPs在小鼠中引发了更有效的免疫反应，表现为比α -Al₂O₃ +UPs分泌更多的IFN -γ。再者，α -Al₂O₃ -UPs在4T1荷瘤小鼠中对肿瘤生长抑制和生存期延长也展现出更强的作用。值得注意的是，与低剂量化疗联合使用时，抗肿瘤效果比单独使用α -Al₂O₃ -UPs时进一步增强。
结论：本研究通过一种新的简单方法制备了一种内置佐剂纳米疫苗，该疫苗有潜在的癌症免疫治疗应用前景，为未来的临床应用奠定了实验基础。
Background and Aim: We have previously identified ubiquitinated proteins (UPs) from.tumor cell lysates as a promising vaccine for cancer immunotherapy in different mouse.tumor models. In this study, we aimed at developing a highly efficient therapeutic adjuvant.built-in nanovaccine (α-Al2O3-UPs) by a simple method, in which UPs from tumor cells.could be efficiently and conveniently enriched by α-Al2O3 nanoparticles covalently coupled.with Vx3 proteins (α-Al2O3-CONH-Vx3)..Methods: The α-Al2O3 nanoparticles were modified with 4-hydroxybenzoic acid followed.by coupling with ubiquitin-binding protein Vx3. It was then used to enrich UPs from 4T1.cell lysate. The stability and the efficiency for the UPs enrichment of α-Al2O3-CONH-Vx3.were examined. The ability of α-Al2O3-UPs to activate DCs was examined in vitro subsequently. The splenocytes from the vaccinated mice were re-stimulated with inactivated tumor.cells, and the IFN-γ secretion was detected by ELISA and flow cytometry. Moreover, the.therapeutic efficacy of α-Al2O3-UPs, alone and in combination with chemotherapy, was.examined in 4T1 tumor-bearing mice..Results: Our results showed that α-Al2O3-UPs were successfully synthesized and abundant.UPs from tumor cell lysate were enriched by the new method. In vitro study showed that.compared to the physical mixture of α-Al2O3 nanoparticles and UPs (α-Al2O3+UPs), α-Al2 O3-UPs stimulation resulted in higher upregulations of CD80, CD86, MHC class I, and MHC.class II on DCs, indicating the higher ability of DC activation. Moreover, α-Al2O3-UPs.elicited a more effective immune response in mice, demonstrated by higher IFN-γ secretion.than α-Al2O3+UPs. Furthermore, α-Al2O3-UPs also exhibited a more potent effect on tumor.growth inhibition and survival prolongation in 4T1 tumor-bearing mice. Notably, when in.combination with low dose chemotherapy, the anti-tumor effect was further enhanced, rather.than using α-Al2O3-UPs alone..Conclusion: This study presents an adjuvant built-in nanovaccine generated by a new.simple method that can be potentially applied to cancer immunotherapy and lays the.experimental foundation for future clinical application.