Specific antibody response of mice after immunization with COS-7 cell derived avian influenza virus (H5N1) recombinant proteins.

Navin Horthongkham, Tananun Srihtrakul, Niracha Athipanyasilp, Sontana Siritantikorn, Wannee Kantakamalakul, Yong Poovorawan, Ruengpung Sutthent
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引用次数: 3

Abstract

To develop avian influenza H5N1 recombinant protein, the hemagglutinin (HA), neuraminidase (NA), matrix (M), and non-structural (NS1) of avian influenza H5N1 isolates from Thailand were engineered to be expressed in prokaryotic (E. coli) and mammalian cell (COS-7) system. The plasmid pBAD-His and pSec-His were used as vectors for these inserted genes. Mice immunized with purified recombinant proteins at concentration 50-250 mug intramuscularly with Alum adjuvant at week 0, week 2, and week 3 showed a good immunogenicity measured by ELISA and neutralization assay. The HA and NS recombinant proteins produced in COS-7 cells can induce specific antibody titer detected by neutralization assay significantly higher than corresponding recombinant proteins produced in E. coli system. The antibody produced in immunized mice could neutralize heterologous avian influenza virus determined by micro-neutralization assay. This study shows that avian influenza virus H5N1 recombinant proteins produced in mammalian cell system were able to induce neutralizing antibody response.

Abstract Image

Abstract Image

COS-7细胞源性禽流感病毒(H5N1)重组蛋白免疫小鼠后的特异性抗体反应
为了开发H5N1禽流感重组蛋白,对泰国H5N1禽流感分离株的血凝素(HA)、神经氨酸酶(NA)、基质(M)和非结构(NS1)进行了工程设计,在原核(大肠杆菌)和哺乳动物细胞(COS-7)系统中表达。质粒pBAD-His和pSec-His分别作为这些插入基因的载体。以50 ~ 250杯浓度的纯化重组蛋白与明矾佐剂分别于第0周、第2周和第3周肌内免疫小鼠,经ELISA和中和实验检测,显示出良好的免疫原性。通过中和试验检测,COS-7细胞中产生的HA和NS重组蛋白可诱导特异性抗体滴度显著高于大肠杆菌系统中产生的相应重组蛋白。微中和试验证实免疫小鼠产生的抗体能中和异源禽流感病毒。本研究表明,在哺乳动物细胞系统中产生的H5N1禽流感病毒重组蛋白能够诱导中和抗体反应。
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