以新型脂肽为佐剂的LP-LNP构建及其对mRNA疫苗的增效作用

曹静文; 迟宇; 李郭成; 程浩; 邓炎; 魏静; 朱吉; 高英英; 李海波

doi:10.16016/j.2097-0927.202402048

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以新型脂肽为佐剂的LP-LNP构建及其对mRNA疫苗的增效作用

基础医学 | 更新时间：2026-01-09

- 以新型脂肽为佐剂的LP-LNP构建及其对mRNA疫苗的增效作用
- Construction of LP-LNP with novel lipopeptides as adjuvants and its enhancing effects on mRNA vaccines
- 陆军军医大学学报 2024年46卷第17期页码：1925-1933
- 作者机构：
  
  1.154007 黑龙江佳木斯，佳木斯大学临床医学院
  2.400038 重庆，陆军军医大学(第三军医大学)药学与检验医学系微生物与生化药学教研室，国家免疫生物制品工程技术研究中心
  3.401320 重庆，重庆医科大学附属巴南医院检验科
- 作者简介：
  
  [高英英，E-mail：63543545@qq.com GAO Yingying, E-mail: 63543545@qq.com
  [李海波，E-mail：lihaibo@tmmu.edu.cn LI Haibo, E-mail: lihaibo@tmmu.edu.cn
- 基金信息：
  
  国家自然科学基金面上项目(32270988);重庆市自然科学基金重点项目(cstc2020jcyj-zdxmX0027)
- DOI：10.16016/j.2097-0927.202402048
  中图分类号： R342.22;R914.2;R979.5
- 收稿：2024-02-19，
  
  修回：2024-04-16，
  
  纸质出版：2024-09-15
- 稿件说明：
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曹静文, 迟宇, 李郭成, 等. 以新型脂肽为佐剂的LP-LNP构建及其对mRNA疫苗的增效作用[J]. 陆军军医大学学报, 2024,46(17):1925-1933.

Jingwen CAO, Yu CHI, Guocheng LI, et al. Construction of LP-LNP with novel lipopeptides as adjuvants and its enhancing effects on mRNA vaccines[J]. Journal of Army Medical University, 2024, 46(17): 1925-1933.
曹静文, 迟宇, 李郭成, 等. 以新型脂肽为佐剂的LP-LNP构建及其对mRNA疫苗的增效作用[J]. 陆军军医大学学报, 2024,46(17):1925-1933. DOI： 10.16016/j.2097-0927.202402048.

Jingwen CAO, Yu CHI, Guocheng LI, et al. Construction of LP-LNP with novel lipopeptides as adjuvants and its enhancing effects on mRNA vaccines[J]. Journal of Army Medical University, 2024, 46(17): 1925-1933. DOI： 10.16016/j.2097-0927.202402048.

摘要

目的

构建以新型脂肽为佐剂的递送系统脂肽-脂质纳米颗粒(lipopeptide-lipid nanoparticle，LP-LNP)，初步探究其对mRNA疫苗的增效作用。

方法

设计并合成2种新型脂肽SS-10和SQ18，微流控技术将编码增强型绿色荧光蛋白(enhanced green fluorescent protein，eGFP)、萤火虫荧光素酶(firefly luciferase，F-luc)和卵清白蛋白(ovalbumin，OVA)的mRNA包载到LP-LNP中，构建以新型脂肽为佐剂的mRNA递送系统；利用动态光散射法表征LP-LNP的粒径、多分散系数；利用HEK-Blue

mTLR2报告细胞检测其Toll样受体2(Toll-like receptors 2

TLR2)的激活作用，筛选最优脂肽掺入比；将优选的LP-LNP-eGFP-mRNA转染至HEK293T细胞，荧光显微镜观察eGFP的表达；利用活体成像技术考察LP-LNP-F-luc-mRNA在小鼠体内的表达水平；流式细胞术评价LP-LNP-OVA-mRNA免疫小鼠后诱导引流淋巴结中树突状细胞(dendritic cells，DCs)成熟水平以及交叉递呈抗原的能力。

结果

脂肽SQ18和SS-10分别以0.50%、0.75%的摩尔比掺入后，得到的LP-LNP平均粒径分别为127和121 nm，平均多分散系数分别为0.11和0.08，包封率均在90%以上，且体外安全性好；该配方条件下激活TLR2的能力显著强于阳性对照Pam

CSK

(

＜0.01)；优选的LP-LNP均可以实现有效的体外转染，其中SQ18以0.50%掺入制备的LP-LNP体内转染效果显著优于传统LNP(

＜0.01)，且可以显著促进引流淋巴结DCs的成熟以及抗原的交叉递呈(

＜0.05)。

结论

以新型脂肽为佐剂的LP-LNP可以增强mRNA的递送能力，进一步提高mRNA疫苗的免疫效果。

Abstract

Objective

To construct lipid nanoparticles (lipopeptide-lipid nanoparticle

LP-LNP) with novel lipopeptides as adjuvants

and initially explore their synergistic effect on mRNA vaccines.

Methods

Two novel lipopeptides

SS-10 and SQ18

were designed and synthesized. Microfluidic technology was used to encapsulate lipopeptides in differ

ent proportions

as well as mRNAs encoding enhanced green fluorescent protein (eGFP)

firefly luciferase (F-luc)

and ovalbumin (OVA) into lipid nanoparticles to construct an mRNA delivery system with novel lipopeptides as adjuvants (LP-LNP). The particle size and polydispersity coefficient of LP-LNP were measured using dynamic light scattering. The activation effect on Toll-like receptors 2 (TLR2) was detected using HEK-Blue

mTLR2 reporter cells to screen the optimal lipopeptide ratio. The preferred LP-LNP-eGFP-mRNA was transfected into HEK293T cells

and the expression of eGFP was observed under a fluorescence microscope.

In vivo

imaging was used to investigate the expression level of LP-LNP-F-luc-mRNA in mice. Flow cytometry was used to evaluate the ability of LP-LNP-OVA-mRNA to induce the maturation of dendritic cells (DCs) in draining lymph nodes and cross-presentation of antigens after immunization.

Results

Lipopeptides SQ18 and SS-10 were incorporated into LNP at 0.50% and 0.75% molar ratios

respectively

to obtain LP-LNP with uniform particle size

high encapsulation efficiency

and good

in vitro

safety. The ability of this formulation to activate TLR2 was significantly stronger than the positive control Pam

CSK

(

0.01). The preferred LP-LNP obtained effective

in vitro

transfection

and LP-LNP prepared with SQ18 at 0.50% molar ratio had significantly better

in vivo

transfection efficiency than traditional LNP (

0.01)

and significantly promoted the maturation of DCs in draining lymph nodes and cross-presentation of antigens (

0.05).

Conclusion

LP-LNP with novel lipopeptides as adjuvants can enhance the delivery capacity of mRNA and further improve the immune effect of mRNA vaccines.

关键词

Keywords

references

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