ABSTRACT
For improving epitope immunogenicity and achieving the co-immunization, late protein 1 (L1) of HPV type 16 (HPV16L1) was selected as the vector to carry the dominant epitope of Toxoplasma gondii because of the shared common population between Toxoplasma gondii and human papillomavirus (HPV). RSepitope-HPV16L1 (RSepitope fused at the "N-terminus" of HPV16L1) and HPV16L1-RSepitope (RSepitope fused at the "C-terminus" of HPV16L1) chimeras were constructed. After transfection of COS-7 cells with the recombinants, Western blot, RT-PCR, and immunofluorescence experiments confirmed that RSepitope-HPV16L1 could successfully express the corresponding mRNA and protein of RSepitope and HPV16L1, but the HPV16L1-RSepitope construct could not. A "prime-boost" immunization program was applied in mice to further evaluate the immune response elicited by the constructs, and the RSepitope-HPV16L1 immunization group produced the most significantly increased humoral and cellular immune responses (the highest RSepitope-specific IgG antibody level and the highest IFN-γ production, respectively), in which both elevated Th1 and Th2 immune responses were obtained. Moreover, the advantage of HPV16L1 as an epitope carrier was remarkable for RSepitope-HPV16L1, which induced a more prominent immunological response than RSepitope alone (without fusion with HPV16L1). Our research indicated that the N-terminus of HPV16L1 could be a better insertion site for enhancing target epitope immunogenicity, and our study offers a design for epitope vaccine of reasonable combination.
Subject(s)
Animals , Mice , Antibody Formation , Epitopes , Immunization , Mice, Inbred BALB C , Toxoplasma , Vaccination , Vaccines, DNAABSTRACT
AIM:To investigate the effect of curcumin derivatives B06(B06) on the synthesis of testosterone from type 2 diabetic rats.METHODS: Male Sprague-Dawley rats were evenly divided into 5 groups randomly: normal control group (C group), high fat group (H group), high fat treatment group (HT group), diabetes mellitus group (D group) and diabetes treatment group ( DT group) .The rats in the later 4 groups were fed with high fat diet, after 4 weeks of high fat diet feeding, the rats from D group and DT group were injected with low dose of streptozotocin intraperitoneally to induce diabetes mellitus, while the rats in HT group and DT group were gavaged with B06 at the dose of 0.2 mg· kg-1 · d-1 for 8 weeks.The blood glucose was detected by glucometer, blood insulin was assayed by ELISA and the insulin resist-ance index was calculated.The morphology of testes were observed by light and transmission electron microscopy.Serum testosterone and estradiol were measured by radioimmunoassay.The protein expression of steroidogenic acute regulatory pro-tein ( StAR) was detected by immunohistochemistry.The mRNA expression of StAR, cholesterol side-chain cleavage en-zyme (P450scc), cytochrome P450 17A1 (P450c17), cytochrome P450 aromatizing enzyme (P450arom), 3β-hydroxyste-roid dehydrogenase ( HSD) and 17β-HSD was detected by RT-PCR.RESULTS:The levels of blood glucose and insulin resistance index were increased in H group and D group, and serum testosterone was decreased, all of which were reversed after the treatment of B06.Testicular seminiferous tubule was distorted, spermatogenic cells were dropped in H group and D group.In addition, leydig cells were found to have swelling mitochondria in H group and D group, endoplasmic reticu-lum was reduced, and there was karyopyknosis accompany with sparse chromatin, all of which were ameliorated by B06. The protein expression of StAR was decreased in D group.The mRNA expression of StAR and P450scc was decreased in H group and D group, all of which were increased in B06 treatment group.There was no significant difference in the mRNA expression of P450c17, P450arom, 3β-HSD and 17β-HSD.CONCLUSION: B06 may increase serum testosterone and relieve the damage of testes from type 2 diabetic rats.B06 improves metabolic disorder by up-regulating mRNA expression of StAR and P450scc.