Leya Tasok, Ahmad Irshad, Sharma Rupam, Tripathi Gayatri, Kurcheti Pani Prasad, Rajendran Kooloth Valappil, Bedekar Megha Kadam
ICAR-Central Institute of Fisheries Education, Mumbai 400061, India; School of Fisheries, Centurion University of Technology and Management, R-Sitapur, Paralakhemundi, Odisha 761211, India.
College of Fisheries Science, Gumla, Birsa Agricultural University, Ranchi 834006, India.
Int J Biol Macromol. 2020 Aug 1;156:928-937. doi: 10.1016/j.ijbiomac.2020.04.048. Epub 2020 Apr 11.
DNA vaccine is an important tool to elicit both humoral and cellular immunity. Present study investigates mucosal immune response of Labeo rohita (15 ± 04 g) to plasmid DNA (pDNA) vaccine macromolecule complexed with nanoparticles (NPs). Poly lactic-co-glycolic acid (PLGA), Chitosan (Chit) and PLGA-Chit-NPs were synthesized by double emulsion solvent evaporation method. Synthesized NPs were complexed with pDNA (pGPD + IFN) vaccine construct. Size and zeta potential of PLGA-NPs, Chit-NPs and PLGA-Chit-NPs-pDNA complex were recorded to be 120 nm and +0.5 mV, 117 nm and +32 mV, 189 nm and +11 mV, respectively. Immunization by immersion was carried out in two groups receiving PLGA-Chit-NPs-pDNA (T1) and PLGA-NPs-pDNA (T2) respectively. After immersion, samples were collected on 0, 2, 4, 7, 15 and 30 days from mucosa-associated lymphoid tissues (MALT) for mRNA expression studies of IgM, IgD and IgZ using qRT-PCR. Significant up-regulation of the mRNA expression of IgM, IgD, and IgT were observed in MALT in immunized fish compared to control. After 30 days post-immunization fish were infected with a virulent strain of Edwardsiella tarda. The highest relative percentage survival was observed in T1 (64.7%) compared to T2. The study showed better efficiency of pDNA-PLGA-Chit-NPs compared to pDNA-PLGA-NPs for inducing adaptive mucosal immunity in fish.
DNA疫苗是引发体液免疫和细胞免疫的重要工具。本研究调查了体重为15±0.4克的露斯塔野鲮对与纳米颗粒（NPs）复合的质粒DNA（pDNA）疫苗大分子的黏膜免疫反应。通过双乳液溶剂蒸发法合成了聚乳酸-乙醇酸共聚物（PLGA）、壳聚糖（Chit）和PLGA-壳聚糖纳米颗粒（PLGA-Chit-NPs）。合成的纳米颗粒与pDNA（pGPD + IFN）疫苗构建体复合。记录到PLGA纳米颗粒、壳聚糖纳米颗粒和PLGA-壳聚糖纳米颗粒-pDNA复合物的大小和zeta电位分别为120纳米和+0.5毫伏、117纳米和+32毫伏、189纳米和+11毫伏。分别在两组中通过浸泡进行免疫，一组接受PLGA-壳聚糖纳米颗粒-pDNA（T1），另一组接受PLGA纳米颗粒-pDNA（T2）。浸泡后，在第0、2、4、7、15和30天从黏膜相关淋巴组织（MALT）收集样本，使用qRT-PCR研究IgM、IgD和IgZ的mRNA表达。与对照组相比，在免疫鱼的MALT中观察到IgM、IgD和IgT的mRNA表达显著上调。免疫后30天，鱼感染了迟缓爱德华氏菌的强毒株。与T2组相比，T1组观察到的相对存活率最高（64.7%）。该研究表明，与pDNA-PLGA纳米颗粒相比，pDNA-PLGA-壳聚糖纳米颗粒在诱导鱼的适应性黏膜免疫方面效率更高。
