[1]付倩梅,唐华明,张 鹏,等.偶联 CD206 抗体载 Fe3O4的 PLGA 纳米微球促进巨噬细胞 M1型极化[J].南方医科大学学报,2020,(02):246-254.[doi:10.12122/j.issn.1673-4254.2020.02.05]
 Anti-CD206 antibody- conjugated Fe3O4-based PLGA nanoparticles selectively promotesM1 polarization of tumor-associated macrophages in mice[J].Journal of Southern Medical University,2020,(02):246-254.[doi:10.12122/j.issn.1673-4254.2020.02.05]
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偶联 CD206 抗体载 Fe3O4的 PLGA 纳米微球促进巨噬细胞 M1型极化()
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《南方医科大学学报》[ISSN:1673-4254/CN:44-1627/R]

卷:
期数:
2020年02期
页码:
246-254
栏目:
出版日期:
2020-02-29

文章信息/Info

Title:
Anti-CD206 antibody- conjugated Fe3O4-based PLGA nanoparticles selectively promotes M1 polarization of tumor-associated macrophages in mice
作者:
付倩梅唐华明张 鹏阙克婷刘作金周 赟
关键词:
PLGA微球纳米氧化铁靶向药物转运肿瘤的免疫治疗
Keywords:
poly (lactic-co-glycolic acid) iron oxide nanoparticles targeted drug delivery tumor immunotherapy
DOI:
10.12122/j.issn.1673-4254.2020.02.05
文献标志码:
A
摘要:
目的 通过构建纳米微球靶向性地提高巨噬细胞中的铁浓度来增强抗肿瘤免疫。方法 采用W/O/W复乳化溶剂扩散法制备CD206单克隆抗体表面修饰的载Fe3O4聚乳酸羟基乙酸(PLGA)纳米微球。使用马尔文粒径检测仪测定微粒直径,Zeta电位 法测定Zeta电位。用铁测定试剂盒测定Fe3O4的包封率。采用免疫荧光实验检测CD206抗体与巨噬细胞的结合及靶向性。Western blot、qRT-PCR检测巨噬细胞的极化指数。并用BALB/C-57小鼠皮下肿瘤模型验证纳米微球促进肿瘤相关巨噬细胞(TAM)的极化状态。结果 纳米微球的平均直径在260~95 nm范围内,Zeta电位值在-19~-33 MV。Fe3O4的包封率在65%~75%。流式细胞术检测CD206单克隆抗体与PLGA微球偶联率为65%~70%,免疫荧光实验证实了PLGA微球与CD206高表达巨噬细胞的靶向结合能力。Western blot和qRT-PCR证实了偶联CD206抗体载Fe3O4的PLGA纳米微球(CD206-Fe3O4-PLGA)和载Fe3O4的PLGA纳米微球(Fe3O4-PLGA)促进TNF-α、iNOS和IL-1β的表达(P<0.05)。小鼠肿瘤模型研究证实CD206-Fe3O4-PLGA纳米颗粒促进TAMs中CD86的表达。结论 PLGA纳米微球具有均匀的粒子的大小及Zeta电位,以及较好的抗体偶联效率及纳米铁包封率,同时偶联CD206的PLGA微球能够较好的靶向结M2型巨噬细胞,并通过释放包被的Fe3O4促进巨噬细胞的M1型极化。本研究为肿瘤的免疫治疗提供了一种潜在的方法。
Abstract:
Objective To enhance the anti-tumor immunity of macrophages by increasing iron concentration in the macrophages using nanospheres. Methods Anti-CD206 antibody-conjugated Fe3O4-based polylactic acid glycolic acid (CD206- Fe3O4-PLGA) nanoparticles were prepared with the W/O/W method. The particle diameter was measured using Malvern particle size detector, the Zeta potential was determined using Zeta potentiometry, and the encapsulation efficiency of Fe3O4 was determined using an iron determination kit. The macrophage-binding and targeting abilities of the conjugated nanoparticles were evaluated using immunofluorescence assay, and the polarization index of macrophages was determined with Western blotting and qRT-PCR. BALB/C-57 mouse models bearing subcutaneous tumors were used to verify the efficacy of the nanoparticles to promote polarization of the tumor-associated macrophages (TAMs). Results The conjugated nanoparticles had a mean diameter of 260-295 nm with Zeta potential values ranging from -19 mV to -33 mV, encapsulation efficiency of Fe3O4 ranging from 65% to 75%, and anti-CD206 conjunction efficiency of 65%-70%. Immunofluorescence assay verified the targeted binding ability of the nanoparticles with M2 macrophages. Western blotting and qRT-PCR confirmed that both CD206-Fe3O4-PLGA and Fe3O4-PLGA nanoparticles promoted the expression of TNF-α, iNOS and IL-1β (P<0.05). In the tumor-bearing mouse models, CD206-Fe3O4-PLGA nanoparticles were confirmed to promote CD86 expression in the TAMs. Conclusion CD206-Fe3O4-PLGA nanoparticles are capable of targeted binding to M2 macrophages and reversing the M2 macrophages to M1 phenotype by releasing coated iron oxide particles.
更新日期/Last Update: 2020-03-14