Construction of the DNA vaccine plasmid against CDR3 region of immunoglobulin heavy chain of the human B-cell lymphoma cell line Namalwa / 中国免疫学杂志

ABSTRACT

Objective:To analyze whether the DNA vaccine against the CDR3 regio n of immunoglobulin heavy chain of the human B-cell lymphoma could elicit the s p ecific idiotypic antibody,the authors established a model with the human B-cell lymphoma c ell line Namalwa.The CDR3 gene fragment of Namalwa membranous immunoglobulin hea vy chain was amplified.The sequenced CDR3 fragment was used as the antigen gene to construct the DNA vaccine plasmid. Methods:The authors acquired the CDR3 gene fragment using Ig superfamily primers by means of RT-PCR and the murine monocyte chemot ic protein(MCP-3) as the adjuvant molecular.The fused gene fragment of CDR3 and MCP-3 was obtained by recombinant PCR and then cloned into the eukaryonic plas mid vector pcDNA3.1 to construct the DNA vaccine plamid.Then the vaccine plasmid was transiently expressed in the eukaryonic cell COS-7. Results:The authors acquired th e DNA vaccine plasmid in which the mIgCDR3 of the Namalwa cell was used as the a ntigen gene by the molecular biology. Conclusion:The transient transfection assa y proved that the recombinant plasmid could express in eukaryonic cells in right way.They have constructed an expression plasmid containing fused MCP3-CDR3 seq uen ce which could be used in further study of DNA vaccine against B-cell lymphoma in vivo. [