Khademi Farzad, Yousefi Arshid, Derakhshan Mohammad, Najafi Adel, Tafaghodi Mohsen
Department of Microbiology, School of Medicine, Ardabil University of Medical Sciences, Ardabil, Iran.
Department of Medical Bacteriology and Virology, Qaem University Hospital, School of Medicine, Mashhad University of Medical Sciences, Mashhad, Iran.
Iran J Basic Med Sci. 2019 Aug;22(8):893-900. doi: 10.22038/ijbms.2019.33962.8079.
A new strategy in recent studies is using effective tuberculosis (TB) subunit vaccines combined with appropriate carriers and adjuvants which have shown promising results in preclinical and clinical studies. The aim of the present study was to evaluate the PLGA:DDA hybrid nanoparticles (NPs) for subcutaneous delivery of a novel multistage subunit vaccine along with MPLA adjuvant against ().
PLGA and PLGA:DDA NPs containing HspX/EsxS fusion protein and MPLA were prepared by double emulsion method (w/o/w). After characterization, these NPs were subcutaneously administered to BALB/c mice aged 6-8 weeks old. Immunogenicity of formulations were assessed by measuring the level of IFN-γ, IL-4, IL-17 and TGF-β cytokines as well as IgG1, IgG2a and IgA antibodies using ELISA.
Both particles had spherical shape and smooth surface with 316.7±12.7 nm in size, surface charge of -33±1.7 mV, and encapsulation efficiency of 92.2±2% for PLGA NPs and 249.7±16.7 nm in size, surface charge of 39±1.8 mV, and encapsulation efficiency of 35.7±1.4% for PLGA:DDA NPs. The highest IFN-γ response and also IgG2a and IgG1 antibodies titers were observed in groups immunized with PLGA:DDA/HspX/EsxS/MPLA and PLGA:DDA/HspX/EsxS/MPLA as booster as well as PLGA:DDA/HspX/EsxS and PLGA:DDA/HspX/EsxS as booster.
With regard to effective induction of IFN-γ and IgG2a immune responses, PLGA:DDA hybrid NP along with MPLA adjuvant have good potentials for improving the immunogenicity of HspX/EsxS multistage subunit vaccine as well as promoting BCG efficacy as a BCG prime-boost.
近期研究中的一种新策略是使用有效的结核亚单位疫苗与合适的载体和佐剂相结合，这在临床前和临床研究中已显示出有前景的结果。本研究的目的是评估聚乳酸-羟基乙酸共聚物：二油酰基磷脂酰乙醇胺（PLGA:DDA）混合纳米颗粒（NPs）用于皮下递送一种新型多阶段亚单位疫苗以及与单磷酰脂质A（MPLA）佐剂联合用于对抗（）。
通过双乳液法（水包油包水）制备含有热休克蛋白X（HspX）/分泌性蛋白EsxS融合蛋白和MPLA的PLGA及PLGA:DDA纳米颗粒。表征后，将这些纳米颗粒皮下注射给6 - 8周龄的BALB/c小鼠。通过酶联免疫吸附测定（ELISA）测量干扰素-γ（IFN-γ）、白细胞介素-4（IL-4）、白细胞介素-17（IL-17）和转化生长因子-β（TGF-β）细胞因子水平以及IgG1、IgG2a和IgA抗体，评估制剂的免疫原性。
两种颗粒均呈球形且表面光滑，PLGA纳米颗粒大小为316.7±12.7 nm，表面电荷为 - 33±1.7 mV，包封率为92.2±2%；PLGA:DDA纳米颗粒大小为249.7±16.7 nm，表面电荷为39±1.8 mV且包封率为35.7±1.4%。在用PLGA:DDA/HspX/EsxS/MPLA和PLGA:DDA/HspX/EsxS/MPLA作为加强免疫以及PLGA:DDA/HspX/EsxS和PLGA:DDA/HspX/EsxS作为加强免疫的组中观察到最高的IFN-γ反应以及IgG2a和IgG1抗体滴度。
关于有效诱导IFN-γ及IgG2a免疫反应，PLGA:DDA混合纳米颗粒与MPLA佐剂具有良好潜力，可提高HspX/EsxS多阶段亚单位疫苗的免疫原性以及作为卡介苗（BCG）的初免-加强免疫方案来提高BCG的效力。
