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Evaluation of antibody response in mice against avian influenza A (H5N1) strain neuraminidase expressed in yeast Pichia pastoris

机译:小鼠巴斯德毕赤酵母中表达的针对禽流感A(H5N1)株神经氨酸酶的抗体反应的评估

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Avian influenza has raised many apprehension in the recent years because of its potential transmitability to humans. With the increasing emergence of drug-resistant avian influenza strains, development of potential vaccines are imperative to manage this disease. Two structural antigens, haemagglutinin and neuraminidase, have been the target candidates for the development of subunit vaccine against influenza. In an effort to develop a faster and economically beneficial vaccine, the neuraminidase gene of a highly pathogenic avian influenza isolate was cloned and expressed in the methylotrophic yeast Pichia pastoris. The recombinant neuraminidase (rNA) antigen was purified, and its bioactivity was analysed. The rNA was found to be functional, as determined by the neuraminidase assay. Four groups of mice were immunized with different concentrations of purified rNA antigen, which were adjuvanted with aluminium hydroxide. The immune response against rNA was analysed by enzyme-linked immunosorbent assay (ELISA) and neuraminidase inhibition assay. The mice groups immunized with 25 e???g and 10 e???g of antigen had a significant immune response against rNA. This method can be utilized for faster and cost-effective development of vaccines for a circulating and newer strain of avian influenza, and would aid in combating the disease in a pandemic situation, in which production time matters greatly.
机译:近年来,禽流感由于其对人类的潜在传播能力而引起了人们的广泛关注。随着耐药禽流感病毒株的出现,必须开发潜在的疫苗来控制这种疾病。两种结构抗原,血凝素和神经氨酸酶已成为开发针对流感的亚基疫苗的目标候选物。为了开发一种更快和经济上有益的疫苗,克隆了高致病性禽流感分离株的神经氨酸酶基因,并在甲基营养酵母巴斯德毕赤酵母中表达。纯化重组神经氨酸酶(rNA)抗原,并对其生物活性进行分析。如神经氨酸酶测定所确定,发现rNA是有功能的。用不同浓度的纯化rNA抗原免疫四组小鼠,将其与氢氧化铝佐剂结合。通过酶联免疫吸附测定(ELISA)和神经氨酸酶抑制测定分析对rNA的免疫应答。用25μg和10μg抗原免疫的小鼠组具有明显的针对rNA的免疫应答。这种方法可用于为循环的和较新的禽流感菌株开发更快,更具成本效益的疫苗,并有助于在生产时间很重要的大流行情况下与该疾病作斗争。

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