ABSTRACT
<p><b>OBJECTIVE</b>The tripartite motif (TRIM) proteins are a family of more than 70 human members, however only a few of them have been well studied. It has been shown that TRIM proteins are involved in various cellular processes such as cell proliferation, differentiation, apoptosis and antiviral defense. The functions of TRIM38 are largely unknown. In this study we explore the effect of TRIM38 on NF-kappaB signaling pathway.</p><p><b>METHODS</b>293T cells were transfected with NF-kappaB-Luc and plasmids expressing TRIM38 and its mutants fused to Flag. 24 h after transfection, cells were harvested and luciferase activities were measured. Data are representative of three independent experiments with triplicate samples. The expression of proteins was analyzed by Western Blot.</p><p><b>RESULTS</b>TRIM38 could activate NF-kappaB signaling pathway. The mutants of TRIM38 affected the function of TRIM38. Only the mutant of SPRY domain deletion had no obviously influence of the function of TRIM38.</p><p><b>CONCLUSION</b>Our study reveals that NF-kappaB is activated in response to TRIM38.</p>
Subject(s)
Humans , Carrier Proteins , Cells, Cultured , NF-kappa B , Physiology , Proteins , Physiology , Signal Transduction , PhysiologyABSTRACT
HIV-1 subtype B gag genes were cloned from the infected paid blood donors in Henan, and the consensus sequence based on these prevalent strains was obtained by aligning. The codons of the consensus gag sequence were modified according to mammalian codon usage. Western blot analysis was used to compare the expression level of wild type and codon-modified gag gene. It was found that the expression level of Gag protein was improved largely by codon-modification. Then the mod. gag gene was inserted into the adenovirus vector and the recombinant adenovirus rAdV-mod. gag was constructed. 10(8) PFU or 10(9) PFU rAdV-mod, gag vaccinated mice twicely could elicit high level Gag-specific CTL responses in immunized mice. In conclusion,the codon modification of gag gene is successful. The recombinant adenovirus vaccine harbouring mod. gag can induce robust Gag-specific CTL immune response in mice.
Subject(s)
Animals , Female , Humans , Mice , Adenoviridae , Genetics , Blotting, Western , Codon , Genetics , Gene Products, gag , Genetics , Allergy and Immunology , Metabolism , HIV-1 , Genetics , Allergy and Immunology , Metabolism , Immunization , Methods , Mice, Inbred BALB C , Mutation , Recombinant Proteins , Allergy and Immunology , Metabolism , Reverse Transcriptase Polymerase Chain Reaction , T-Lymphocytes, Cytotoxic , Allergy and ImmunologyABSTRACT
<p><b>OBJECTIVE</b>To study the immune effect of a chimeric adenovirus type 5 vector with type 35 fiber (rAd5/F35) vaccine in BALB/c mice.</p><p><b>METHODS</b>The expression of HIV Gag protein was determined using indirect immunofluorescent staining. The rAd5/F35-mod.gag vector was injected intramuscularly to mice. The IgG antibody was detected by ELISA and CTL response was detected by intracellular cytokine stain assay.</p><p><b>RESULTS</b>The rAd5/F35-mod.gag vector could express HIV Gag protein in vitro and generate strong HIV-specific immune responses in vivo. But anti-Ad5 immunity could limit its immunogenicity in vivo.</p><p><b>CONCLUSION</b>The rAd5/F35-mod.gag vector can elicit specific CTL response and IgG antibody in animal model. In mice with high Ad5 vector-specific immunity, Ad5/F35-mod.gag showed lower level of Gag specific CTL and antibody response than in mice without pre-existing adenovirus type 5 immunity. The results indicated that fiber exchange alone does not evade pre-existing Ad5 immunity.</p>