Construction and expression of a plasmid encoding B-lymphocyte chemoattractant CXCL13

Objectives: As a B-lymphocyte chemoattractant, CXCL13 may provide “dangerous signal” to immune system and may be used as an adjuvant to enhance the immune responses induced by anti-caries DNA vaccine. Here we report a plasmid encoding CXCL13 gene and detect its expression in eukaryotic cells, which will be used in the future to enhance the efficacy of anti-caries DNA vaccine. Methods: The CXCL13 gene fragment was generated by reverse transcript-PCR from splenic cells and then inserted into pMD18-T vector. The gene fragment was sequenced and then cloned into pVAX1 to get the recombinant plasmid pCXCL13/VAX. 293 cells were transfected with pCXCL13/VAX. The expression of recombinant CXCL13 was detected by immunohistochemitry. Results: Electrophoresis and sequencing results showed the expected fragment was obtained. Positive staining cells were found in the cytoplasm of pCXCL13/VAX-transfected cells. Conclusions: pCXCL13/VAX was constructed successfully and could express recombinant protein in 293 cells. The further study will be done to evaluate the ability of CXCL13 to enhance the immune responses induced by anti-caries DNA vaccine. The study was supported by National Natural Scientific Foundation of China. 304161099