Pereira Daniele R, Pérez-Betancourt Yunys, Távora Bianca C L F, Magalhães Geraldo S, Carmona-Ribeiro Ana Maria, Faquim-Mauro Eliana L
Laboratory of Immunopathology, Butantan Institute, São Paulo 05585-000, Brazil.
Department of Immunology, Institute of Biomedical Sciences, University of São Paulo, São Paulo 04021-001, Brazil.
Vaccines (Basel). 2025 Jan 16;13(1):76. doi: 10.3390/vaccines13010076.
: Cationic polymers were shown to assemble with negatively charged proteins yielding nanoparticles (NPs). Poly-diallyl-dimethyl-ammonium chloride (PDDA) combined with ovalbumin (OVA) yielded a stable colloidal dispersion (OVA/PDDA-NPs) eliciting significant anti-OVA immune response. Dendritic cells (DCs), as sentinels of foreign antigens, exert a crucial role in the antigen-specific immune response. Here, we aimed to evaluate the involvement of DCs in the immune response induced by OVA/PDDA. : In vivo experiments were used to assess the ability of OVA/PDDA-NPs to induce anti-OVA antibodies by ELISA, as well as plasma cells and memory B cells using flow cytometry. Additionally, DC migration to draining lymph nodes following OVA/PDDA-NP immunization was evaluated by flow cytometry. In vitro experiments using bone marrow-derived DCs (BM-DCs) were used to analyze the binding and uptake of OVA/PDDA-NPs, DC maturation status, and their antigen-presenting capacity. Our data confirmed the potent effect of OVA/PDDA-NPs inducing anti-OVA IgG1 and IgG2a antibodies with increased CD19CD138 plasma cells and CD19CD38CD27 memory cells in immunized mice. OVA/PDDA-NPs induced DC maturation and migration to draining lymph nodes. The in vitro results showed higher binding and the uptake of OVA/PDDA-NPs by BM-DCs. In addition, the NPs were able to induce the upregulation of costimulatory and MHC-II molecules on DCs, as well as TNF-α and IL-12 production. Higher OVA-specific T cell proliferation was promoted by BM-DCs incubated with OVA/PDDA-NPs. : The data showed the central role of DCs in the induction of antigen-specific immune response by OVA-PDDA-NPs, thus proving that these NPs are a potent adjuvant for subunit vaccine design.
阳离子聚合物已被证明可与带负电荷的蛋白质组装形成纳米颗粒(NPs)。聚二烯丙基二甲基氯化铵(PDDA)与卵清蛋白(OVA)结合产生了稳定的胶体分散体(OVA/PDDA-NPs),引发了显著的抗OVA免疫反应。树突状细胞(DCs)作为外来抗原的哨兵,在抗原特异性免疫反应中发挥着关键作用。在此,我们旨在评估DCs在OVA/PDDA诱导的免疫反应中的作用。
体内实验用于通过ELISA评估OVA/PDDA-NPs诱导抗OVA抗体的能力,以及使用流式细胞术评估浆细胞和记忆B细胞。此外,通过流式细胞术评估OVA/PDDA-NP免疫后DC向引流淋巴结的迁移。使用骨髓来源的DCs(BM-DCs)进行的体外实验用于分析OVA/PDDA-NPs的结合和摄取、DC成熟状态及其抗原呈递能力。我们的数据证实了OVA/PDDA-NPs在免疫小鼠中诱导抗OVA IgG1和IgG2a抗体的强大作用,同时增加了CD19CD138浆细胞和CD19CD38CD27记忆细胞。OVA/PDDA-NPs诱导DC成熟并迁移至引流淋巴结。体外结果显示BM-DCs对OVA/PDDA-NPs有更高的结合和摄取。此外,这些纳米颗粒能够诱导DC上共刺激分子和MHC-II分子的上调,以及TNF-α和IL-12的产生。与OVA/PDDA-NPs孵育的BM-DCs促进了更高的OVA特异性T细胞增殖。
数据显示DCs在OVA-PDDA-NPs诱导的抗原特异性免疫反应中起核心作用,从而证明这些纳米颗粒是亚单位疫苗设计的有效佐剂。