[Frontiers in Bioscience 11, 121-125, January 1, 2006]

Construction and direct immunization of eukaryotic expression plasmid encoding human soluble B lymphocyte stimulator

Guangyu Chen 1, Hongwu Du 2, Shanyun Peng 1, Donggang Xu 1, and Jiaxi Wang 1

1 Laboratory of Molecular Genetics, Beijing Institute of Basic Medical Sciences, Beijing, China, 2 Department of Biological Sciences and Biotechnology, Tsinghua University, Beijing, China

TABLE OF CONTENTS

1. Abstract
2. Introduction
3. Materials and methods
3.1. Antibodies
3.2. Plasmids construction
3.3. Enzyme-linked immunosorbent assay (ELISA)
3.4. Statistical analysis
4. Results
4.1. Production of the recombinant BLyS
4.2. Construction of eukaryotic expression plasmids of BLyS
4.3. Immunological effects of administered eukaryotic expression plasmid of BLyS on the expression of human BLyS, the type of Igs anti-recombinant protein and serum immunoglobulin in BALB/c mice
5. Discussion
6. Acknowledgments
7. References

1. ABSTRACT

B lymphocyte stimulator of the tumor-necrosis-factor family (BLyS) enhances B cell survival, a function indispensable to B cell maturation. This factor plays a crucial role in enhancing immune responses. Here, to study the primary immune effect of BLyS gene in animals, we constructed recombinant eukaryotic expression plasmid of the human soluble BLyS by using expression vector pcDNA3.1 (-). The recombinant plasmid pcDNA3.1BLyS was injected subcutaneously into BALb/C mice and this administration induced the sustained expression of human BLyS and specific IgG against the recombinant BLyS protein. This study shows that recombinant eukaryotic expression plasmid, pcDNA3.1BLyS, might be used in human gene therapy.