Construction Of Cysticercus Cellulosae AgB Gene Amino-terminal,Carboxyl-terminal Fragment And Porcine CD58 Gene Coexpression DNA Vaccines And Elementary Analyses Of Their Immunological Activity

Inoculation of cysticercus cellulosae AgB recombinant or DNA vaccine can efficiently prevent mouse or pig from infecting of Taenia solium has been proved as many researches in both China and abroad. In this study, total RNA was extracted from peripheral blood mononuclear cells (PBMC) stimulated by ConA, and then an 800bp specific fragment was amplified by reverse transcription-polymerase chain reaction (RT-PCR) using specific primers, and cloned into the vectors later. The positive clones contain the complete open reading frame (ORF) of the CD58 gene was identified by restriction endonuclease analysis, polymerase chain reaction (PCR) and sequencing. Compared the nucleotide and amino acid sequences with the sequences published in GenBank, the homogeneity was 99.4% and 99.2% respectively. Firstly CD58 was inserted into the control of EF-1α promoter of eukaryotic expression vector pBudCE4.1, which contains two promoters, to construct expression plasmid vector PCD, and then the genes of AgB amino-terminal and carboxyl-terminal fragment protein(AgB-N 808bp,AgB-C 943bp)were inserted into the control of CMV promoter of plasmid vector PCD to construct eukaryotic expression recombinant plasmids PBN and PBC respectively, while costimulatory molecule porcine CD58 as immunoadjuvant gene. 4-week-old female healthy BALB/c mice were respectively immunized with PBN and PBC plasmids, and a booster vaccination was given two weeks later. Kinetic tests were carried out by ELISA and MTT assays. Results show that humoral and cellular immunoresponse were not only elicited by PBN and PBC DNA vaccines, but also humoral immunoresponse elicited by PBC was stronger than that by PBN, and cellular immunoresponse elicited by PBN was stronger than that by PBC. Briefly, DNA vaccines can induce stronger cellular immunoresponse and weaker humoral immunoresponse than cysticercus cellulosae purified antigen dose.