ABSTRACT
Objective:To analyze the immune response in mice after immunization with vaccine of rAd5F35-SIVenvT in combination with rMVA-SIVenvT to evaluate the efficacy of different immunization strategies.Methods:Two recombinant viruses were identified in vitro by PCR and Western blot. The BALB/c mice were immunized with homologous and heterologous immune strategies. The numbers of splenic lymphocytes secreting IFN-γ were measured by ELISPOT assay, meanwhile SIV gp120 antibody titer were measured by ELISA assay. Results:SIVenvT protein was expressed effectively by rAd5F35-SIVenvT and rMVA-SIVenvT in HEK293 cells. The specific immune response reached its peak at 4-week post first immunization, then decreased. SIV Env specific cellular immune response and SIV gp120 specific antibody could be detected at 4-16 weeks post first immunization. The specific cellular response was significant stronger in heterologous immunization group than homologous group at 4 week and 16 week. Furthermore, heterologous immunization induced significant higher titer of SIV gp120 antibody at 4 week than homologous group.Conclusions:Specific immune response induced by rAd5F35-SIVenvT in combination with rMVA-SIVenvT was stronger than homologous vector immunization. The results provided references for further study in nonhuman primates.
ABSTRACT
Objective To explore the structure of ribosomal protein S7 ( RPS7) gene of giant panda ( Ailuropoda melanoleuca) and investigate its homologies with other already reported sequences,including Homo sapiens,Mus musculus,Rattus norvegicus and Bos taurus. Methods The cDNA of RPS7 was cloned from the giant panda by RT-PCR. The sequence data were analyzed by GenScan software. Blast 2. 1 was used to study the homology of the obtained RPS7 sequence with the gene sequences of other species; Open reading frame ( ORF) of the DNA sequence was searched using ORF finder software; Protein structure of the RPS7 sequence cloned was deduced using Predict Protein software. Results The full length of the sequence fragment was 589 bp containing an ORF of 585 bp. The deduced protein sequence showed that the protein was composed of 194 ami- no acids and its estimated molecular weight was 22. 126 85 ?103 with a pI of 10. 09. There were 7 different pat- terns of functional sites: one N-glycosylation site; two cAMP and cGMP-dependent kinase phosphorylation sites; four casein kinase C phosphorylation sites; one casein kinase Ⅱ phosphorylation site; two N-myristoylation sites; two amidation sites and one ribosomal protein S7e signature site in the RPS7 protein. Further analysis indicated that the sequence of RPS7 and the protein encoded were highly homologous to some mammals reported.Conclusion The complete coding sequence of RPS7 gene has been cloned through RT-PCR successfully, which is the first report on the RPS7 gene from the giant panda.