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

Navin Horthongkham,Sontana Siritantikorn,Tananun Srihtrakul,Wannee Kantakamalakul,Ruengpung Sutthent,Yong Poovorawan,Niracha Athipanyasilp

doi:10.1186/1476-8518-5-10

Navin Horthongkham, Sontana Siritantikorn + Show 5 more

Open Access

https://doi.org/10.1186/1476-8518-5-10

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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 μg 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.

Highlights

From January 2004, the pandemic of highly pathogenic avian influenza H5N1 (AI) in poultry and human had started from 9 Asian countries, such as Cambodia, China, Indonesia, Japan, Laos, Malaysia, South Korea, Thailand, and Vietnam [1]
Mice were immunized three times intramuscularly (IM) at 1-week interval with 200 μl doses of 50, 100, 150, 200, and 250 μg of rHA, rNA, rNS1, and rM protein produced in E. coli and COS-7 cells plus an emulsion prepared with Alum adjuvant
The mammalian COS-7 cell system has successfully used as host for the efficient production of avian influenza virus (H5N1) proteins including hemagglutinin (HA), neuraminidase (NA), matrix (M), and non-structural (NS1) proteins with yield of 0.05 mg per 100 ml cells

Summary

Introduction

From January 2004, the pandemic of highly pathogenic avian influenza H5N1 (AI) in poultry and human had started from 9 Asian countries, such as Cambodia, China, Indonesia, Japan, Laos, Malaysia, South Korea, Thailand, and Vietnam [1]. The purified proteins, rHA5, rNA1, rNS1, and rM, produced from E. coli and COS-7 cellls, were administered in mice in combination with adjuvant, was capable of eliciting antibody specific for avian influenza virus, detected by ELISA and neutralizing antibody assay

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