自组装纳米颗粒肿瘤疫苗OVA<sub>257-264</sub>-mi3的构建及其保护效果评价

陈源; 高晨; 李宇航; 崔致远; 程新; 张怡; 于博; 顾江; 杨宪

doi:10.16016/j.2097-0927.202401050

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自组装纳米颗粒肿瘤疫苗OVA_257-264-mi3的构建及其保护效果评价

基础医学 | 更新时间：2026-01-09

- 自组装纳米颗粒肿瘤疫苗OVA_257-264-mi3的构建及其保护效果评价
- Construction of self-assembled nanoparticle tumor vaccine OVA_257-264-mi3 and evaluation of its protective efficacy
- 陆军军医大学学报 2024年46卷第12期页码：1361-1368
- 作者机构：
  
  1.401331 重庆，重庆师范大学生命科学学院
  2.400038 重庆，陆军军医大学(第三军医大学)：药学与检验医学系
  3.400038 重庆，陆军军医大学(第三军医大学)：第一附属医院骨科
- 作者简介：
  
  [顾江，E-mail：jianggu2012@163.com GU Jiang, E-mail: jianggu2012@163.com
  [杨宪，E-mail：xianyang2015@163.com YANG Xian, E-mail: xianyang2015@163.com
- 基金信息：
  
  国家重点研发计划(2021YFC2301405)
- DOI：10.16016/j.2097-0927.202401050
  中图分类号： R392-33;R392.7;R979.19
- 收稿：2024-01-14，
  
  修回：2024-02-25，
  
  纸质出版：2024-06-30
- 稿件说明：
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陈源, 高晨, 李宇航, 等. 自组装纳米颗粒肿瘤疫苗OVA_257-264-mi3的构建及其保护效果评价[J]. 陆军军医大学学报, 2024,46(12):1361-1368.

Yuan CHEN, Chen GAO, Yuhang LI, et al. Construction of self-assembled nanoparticle tumor vaccine OVA_257-264-mi3 and evaluation of its protective efficacy[J]. Journal of Army Medical University, 2024, 46(12): 1361-1368.
陈源, 高晨, 李宇航, 等. 自组装纳米颗粒肿瘤疫苗OVA_257-264-mi3的构建及其保护效果评价[J]. 陆军军医大学学报, 2024,46(12):1361-1368. DOI： 10.16016/j.2097-0927.202401050.

Yuan CHEN, Chen GAO, Yuhang LI, et al. Construction of self-assembled nanoparticle tumor vaccine OVA_257-264-mi3 and evaluation of its protective efficacy[J]. Journal of Army Medical University, 2024, 46(12): 1361-1368. DOI： 10.16016/j.2097-0927.202401050.

摘要

目的

构建SpyCatcher-mi3纳米颗粒疫苗递送载体，评价其在增强卵清蛋白CD8

T细胞表位肽OVA

257-264

免疫原性和在小鼠荷瘤模型中免疫保护效果。

方法

SpyCatcher-mi3蛋白由大肠杆菌表达，依次经亲和层析、阴离子交换层析纯化得到；合成OVA

257-264

-SpyTag多肽，通过SpyTag/SpyCatcher蛋白质连接系统构建纳米颗粒OVA

257-264

-mi3；通过细胞溶血实验和细胞毒性实验评价OVA

257-264

-mi3体外安全性，记录小鼠体重变化和镜检脏器组织切片HE染色评价OVA

257-264

-mi3体内安全性；将6~8周龄，18~20 g，SPF级，雌性C57BL/6小鼠按随机数字表法分为OVA

257-264

-mi3组、OVA

257-264

组和Control组，每组14只，分别于第0、14、28天免疫小鼠，末次免疫后第14天通过ELISpot检测其脾脏淋巴细胞中IFN-γ分泌细胞数量评价其免疫原性；在小鼠荷瘤模型上通过核磁共振成像等手段评价纳米颗粒疫苗OVA

257-264

-mi3的保护效果。

结果

SpyCatcher-mi3和OVA

257-264

-mi3均自组装形成均一稳定的纳米颗粒，平均粒径约为43.8 nm和91.3 nm；OVA

257-264

-mi3在体内外均具有良好的安全性；OVA

257-264

-mi3组小鼠每1×10

个脾脏淋巴细胞中IFN-γ分泌细胞数量达253，显著高于OVA

257-264

组(

＜0.05)；OVA

257-264

-mi3组小鼠第22天荷瘤体积约151.1 mm

，显著小于OVA

257-264

组(

＜0.05)，且观察期内生存率达60%，显著高于OVA

257-264

组(

＜0.05)。

结论

成功构建了纳米颗粒疫苗OVA

257-264

-mi3，其增强肿瘤抗原表位肽OVA

257-264

免疫原性，在小鼠荷瘤模型中保护效果良好，为肿瘤新抗原疫苗的研究提供了理论基础。

Abstract

Objective

To construct SpyCatcher-mi3 nanoparticle vaccine delivery vectors

evaluate their role in enhancing the immunogenicity of the ovalbumin CD8

T-cell epitope peptide

OVA

257-264

and determine its protective effect in a model which mice were immunized and subcutaneously challenged with E.G7-OVA tumor cells.

Methods

SpyCatcher-mi3 proteins were expressed by

E.coli

and purified by affinity chromatography and anion exchange chromatography sequentially. OVA

257-264

-SpyTag peptide was obtained by synthesis. The OVA

257-264

-mi3 nanoparticles were produced by the SpyTag/SpyCatcher system. The toxicity of OVA

257-264

-mi3 was evaluated using hemolysis assay

CCK-8 assay and mouse experiment. A total of 42 female SPF-grade C57BL/6 mice (6~8 weeks old

18~20 g) were randomly divided into OVA

257-264

-mi3

OVA

257-264

and control groups

with 14 mice in each group. Then the mice in each group were immunized on days 0

14 and 28. In 14 d after the last immunization

the amounts of spot-forming cells (SFCs

indicating IFN-γ secreting cells in splenic lymphocytes) were determined using ELISpot assay to evaluate their immunogenicity. After the immunized mice were subcutaneously implanted with E.G7-OVA

tumor cells

the antitumor effect of the vaccine in prophylactic xenograft tumor model was evaluate by observing tumor volumes with a caliper and tumor growth with MRI.

Results

Both SpyCatcher-mi3 and OVA

257-264

-mi3 could be self-assembled to form homogeneous and stable nanoparticles

with an average particle size of about 43.8 and 91.3 nm

respectively. The OVA

257-264

-mi3 was safe for

in vitro

and

in vivo

toxicity evaluation. The number of IFN-γ secreting cells per 1×10

splenic lymphocytes reached 253 in the OVA

257-264

-mi3 group of mice

significantly higher than that in the OVA

257-264

group and the Control group (

0.05). The tumor volume of mice in the OVA

257-264

-mi3 group was about 151.1 mm

on day 22

which was significantly smaller than that of the OVA

257-264

group and the Control group (

0.05)

and the survival rate during the observation period reached 60%

which was significantly higher than that of the OVA

257-264

groups (

0.05).

Conclusion

Nanoparticle vaccine OVA

257-264

-mi3 is successfully constructed

and it shows enhancing effect on the immunogenicity of the antigen epitope peptide

and exerts protective effect on prophylactic xenograft tumor model

providing a theoretical basis for the research of tumor neoantigen vaccines.

关键词

Keywords

references

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自组装纳米颗粒肿瘤疫苗OVA257-264-mi3的构建及其保护效果评价

Construction of self-assembled nanoparticle tumor vaccine OVA257-264-mi3 and evaluation of its protective efficacy

DOI：10.16016/j.2097-0927.202401050

摘要

Abstract

关键词

Keywords

references

自组装纳米颗粒肿瘤疫苗OVA_257-264-mi3的构建及其保护效果评价

Construction of self-assembled nanoparticle tumor vaccine OVA_257-264-mi3 and evaluation of its protective efficacy