Braz Luis, Grenha Ana, Ferreira Domingos, Rosa da Costa Ana M, Gamazo Carlos, Sarmento Bruno
CIQA-Centre of Research in Chemistry of Algarve, Faculty of Sciences and Technology, Campus de Gambelas, 8005-139 Faro, Portugal; School of Health-University of Algarve, Avenida Dr. Adelino da Palma Carlos, 8000-510 Faro, Portugal; Department of Pharmaceutical Technology, Faculty of Pharmacy, University of Porto, Rua de Jorge Viterbo Ferreira n.° 228, 4050-313 Porto, Portugal; CBMR-Centre for Biomedical Research, University of Algarve, Faculty of Sciences and Technology, Campus de Gambelas, 8005-139 Faro, Portugal.
CBMR-Centre for Biomedical Research, University of Algarve, Faculty of Sciences and Technology, Campus de Gambelas, 8005-139 Faro, Portugal; CCMAR-Centre for Marine Sciences, University of Algarve, Campus de Gambelas, 8005-139 Faro, Portugal.
Int J Biol Macromol. 2017 Mar;96:786-797. doi: 10.1016/j.ijbiomac.2016.12.076. Epub 2016 Dec 31.
This work proposes the design of nanoparticles based on locus bean gum (LBG) and chitosan to be used as oral immunoadjuvant for vaccination purposes. LBG-based nanoparticles were prepared by mild polyelectrolyte complexation between chitosan (CS) and a synthesized LBG sulfate derivative (LBGS). Morphological characterization suggested that nanoparticles present a solid and compact structure with spherical-like shape. Sizes around 180-200nm and a positive surface charge between +9mV and +14mV were obtained. CS/LBGS nanoparticles did not affect cell viability of Caco-2 cells after 3h and 24h of exposure when tested at concentrations up to 1.0mg/mL. Two model antigens (a particulate acellular extract HE of Salmonella enterica serovar Enteritidis, and ovalbumin as soluble antigen) were associated to CS/LBGS nanoparticles with efficiencies around 26% for ovalbumin and 32% for HE, which resulted in loading capacities up to 12%. The process did not affect the antigenicity of the associated antigens. BALB/c mice were orally immunized with ovalbumin-loaded nanoparticles (100μg), and results indicate an adjuvant effect of the CS/LBGS nanoparticles, eliciting a balanced Th1/Th2 immune response. Thus, CS/LBGS nanoparticles are promising as antigen mucosal delivery strategy, with particular interest for oral administration.
本研究提出了基于刺槐豆胶（LBG）和壳聚糖设计纳米颗粒，用作疫苗接种的口服免疫佐剂。通过壳聚糖（CS）与合成的硫酸化刺槐豆胶衍生物（LBGS）之间的温和聚电解质络合制备了基于LBG的纳米颗粒。形态学表征表明，纳米颗粒呈现出固体且致密的结构，形状类似球形。获得了约180 - 200nm的尺寸以及 +9mV至 +14mV的正表面电荷。当在高达1.0mg/mL的浓度下测试时，CS/LBGS纳米颗粒在暴露3小时和24小时后对Caco - 2细胞的细胞活力没有影响。两种模型抗原（肠炎沙门氏菌肠炎血清型的颗粒性无细胞提取物HE和作为可溶性抗原的卵清蛋白）与CS/LBGS纳米颗粒结合，卵清蛋白的结合效率约为26%，HE的结合效率约为32%，这导致负载能力高达12%。该过程不影响相关抗原的抗原性。用负载卵清蛋白的纳米颗粒（100μg）对BALB/c小鼠进行口服免疫，结果表明CS/LBGS纳米颗粒具有佐剂作用，可引发平衡的Th1/Th2免疫反应。因此，CS/LBGS纳米颗粒作为抗原黏膜递送策略具有潜力，尤其在口服给药方面具有特别的意义。
