Babapoor Sankhiros, Neef Tobias, Mittelholzer Christian, Girshick Theodore, Garmendia Antonio, Shang Hongwei, Khan Mazhar I, Burkhard Peter
Department of Pathobiology and Veterinary Science, University of Connecticut, 61 North Eagleville Road, Storrs, CT 06268, USA.
Influenza Res Treat. 2011;2011:126794. doi: 10.1155/2011/126794. Epub 2012 Jan 12.
Using peptide nanoparticle technology, we have designed two novel vaccine constructs representing M2e in monomeric (Mono-M2e) and tetrameric (Tetra-M2e) forms. Groups of specific pathogen free (SPF) chickens were immunized intramuscularly with Mono-M2e or Tetra-M2e with and without an adjuvant. Two weeks after the second boost, chickens were challenged with 107.2 EID50 of H5N2 low pathogenicity avian influenza (LPAI) virus. M2e-specific antibody responses to each of the vaccine constructs were tested by ELISA. Vaccinated chickens exhibited increased M2e-specific IgG responses for each of the constructs as compared to a non-vaccinated group. However, the vaccine construct Tetra-M2e elicited a significantly higher antibody response when it was used with an adjuvant. On the other hand, virus neutralization assays indicated that immune protection is not by way of neutralizing antibodies. The level of protection was evaluated using quantitative real time PCR at 4, 6, and 8 days post-challenge with H5N2 LPAI by measuring virus shedding from trachea and cloaca. The Tetra-M2e with adjuvant offered statistically significant (P < 0.05) protection against subtype H5N2 LPAI by reduction of the AI virus shedding. The results suggest that the self-assembling polypeptide nanoparticle shows promise as a potential platform for a development of a vaccine against AI.
利用肽纳米颗粒技术,我们设计了两种新型疫苗构建体,分别以单体(Mono-M2e)和四聚体(Tetra-M2e)形式呈现M2e。将特定病原体-free(SPF)鸡分组,分别用Mono-M2e或Tetra-M2e进行肌肉注射免疫,同时设置有无佐剂的组。第二次加强免疫两周后,用107.2 EID50的H5N2低致病性禽流感(LPAI)病毒对鸡进行攻毒。通过ELISA检测每种疫苗构建体的M2e特异性抗体反应。与未接种疫苗的组相比,接种疫苗的鸡对每种构建体的M2e特异性IgG反应均有所增加。然而,疫苗构建体Tetra-M2e与佐剂一起使用时引发的抗体反应明显更高。另一方面,病毒中和试验表明免疫保护不是通过中和抗体实现的。在H5N2 LPAI攻毒后第4、6和8天,通过测量气管和泄殖腔的病毒排出量,使用定量实时PCR评估保护水平。含佐剂的Tetra-M2e通过减少禽流感病毒排出量,对H5N2 LPAI亚型提供了具有统计学意义(P < 0.05)的保护。结果表明,自组装多肽纳米颗粒有望成为开发禽流感疫苗的潜在平台。