Construction Of Rabies Virus Glycoprotein DNA Vaccine And Immune Response In Mice Induced By The Vaccine

Preparing the plasmid of the rabies glycoprotein DNA vaccine and inducethe immune responses in NIH mice,to lay a foundation of a new rabies vaccine.Isolated the G gene of the CTN strain,the site II part of G of the SRV9strain(aa 1-252) and the site III part of G of the CTN strain(aa 253-504) byRT-PCR.Constructing a chimeric rabies G genes,which contains a frame fusion ofaa 1 to 252 from G SRV9 with aa 253 to 504 from CTN.The homogeneous andchimeric rabies G genes were introduced into the eukaryotic expression vectorpcDNA3.1. The inserted foreign gene subtype was determined throughsequence analysis. Plsmids extracted large quantities by alkali lysis andpurified by sepharose 4FF chromatography technique. We injected theplasmids intramuscularly directly into NIH mice. ELISA detected the inducedhumoral immunal response, and cellular immunal response was evaluated byspleen lymphocyte cells transformation experiment and IL-2 activity detectionby CTLL-2 cell line/MTT method.The titers of rabies antibodies were detected in the serum of immunizedmice by plsmids pcDNA/CTN and pcDNA/SRV9CTN DNA vaccineimmunized group;the OD450 values were0.426 and 0.384 respectively. Themean value of ConA stimulated indexes were 7.140 ± 0.5613 in thepcDNA/CTN DNA vaccine immunized mice spleen cells, while in thepcDNA/SRV9CTN and pcDNA3.1 control groups, the mean values were6.476±0.5436 and 1.786±0.528 respectively. The activities of IL-2 canreach 1.812±0.068IU/ml in the pcDNA/CTN immunized groups, while inthepcDNA/SRV9CTN and pcDNA3.1 immunized groups, the IL-2 activitieswere 1.614±0.062IU/ml and 0.163±0.031IU/ml respectively.The plasmidpcDNA/CTN protected 75% of NIH mice against CVS.Under the sameconditions ,pcDNA/SRV9CTN and pcDNA3.1 protected 65%and 25% of NIHmice against CVS respectively.