ABSTRACT
To optimize the immunization strategy against HIV-1, a DNA vaccine was combined with a recombinant vaccinia virus (rTV) vaccine and a protein vaccine. Immune responses against HIV-1 were detected in 30 female guinea pigs divided into six groups. Three groups of guinea pigs were primed with HIV-1 DNA vaccine three times, boosted with rTV at week 14, and then boosted with gp140 protein at intervals of 4, 8 or 12 weeks. Simultaneously, the other three groups of animals were primed with rTV vaccine once, and then boosted with gp140 after 4, 8 or 12 weeks. The HIV-1 specific binding antibody and neutralizing antibody, in addition to the relative affinity of these antibodies, were detected at different time points after the final administration of vaccine in each group. The DNA-rTV-gp140 immune regimen induced higher titers and affinity levels of HIV-1 gp120/gp140 antibodies and stronger V1V2-gp70 antibodies than the rTV-gp140 regimen. In the guinea pigs that underwent the DNA-rTV-gp140 regimen, the highest V1V2-gp70 antibody was induced in the 12-week-interval group. However, the avidity of antibodies was improved in the 4-week-interval group. Using the rTV-gp140 immunization strategy, guinea pigs boosted at 8 or 12 weeks after rTV priming elicited stronger humoral responses than those boosted at 4 weeks after priming. In conclusion, this study shows that the immunization strategy of HIV-1 DNA vaccine priming, followed by rTV and protein vaccine boosting, could strengthen the humoral response against HIV-1. Longer intervals were better to induce V1V2-gp70-specific antibodies, while shorter intervals were more beneficial to enhance the avidity of antibodies.
Subject(s)
Animals , Female , Humans , AIDS Vaccines , Genetics , Allergy and Immunology , DNA, Viral , Genetics , Allergy and Immunology , Guinea Pigs , HIV Infections , Allergy and Immunology , Virology , HIV-1 , Genetics , Allergy and Immunology , Immunization , Methods , Vaccines, DNA , Genetics , Allergy and Immunology , Vaccinia virus , Genetics , Allergy and Immunology , env Gene Products, Human Immunodeficiency Virus , Genetics , Allergy and ImmunologyABSTRACT
<p><b>AIM</b>To investigated the effect of estrogen on global myocardial ischemia/reperfusion (I/R) injury in ovariectomized (Ovx) rats.</p><p><b>METHODS</b>Sprague-Dawley rats were randomly repartitioned into three groups including sham-operated(Sham), ovariectomized (Ovx), or ovariectomized and then given 17beta-estradiol (Ovx + E2). Hearts were excised, mounted on the Langendorff. After the initial stabilization period, all of the three group hearts were randomly divided into normal perfusion subgroup (Control) and I/R perfusion subgroups. Control, perfused for 60 min after stabilization. I/R perfusion subgroups divided into 10 min I + 30 min R, 20 min I + 30 min R, 30 min I + 0 min R, 30 min I + 5 min R, 30 min I + 15 min R and 30 min I + 30 min R. And then, every group hearts were isolated into the single cardiomyocyte. The cardiomyocytes basal contraction and isoproterenol(ISO) stimulation contraction were measured. The viability and yield of cardiomyocytes were counted. LDH and CK concentrations in coronary effluent were assayed with assay kit.</p><p><b>RESULTS</b>The viability and yield of cardiomyocytes were significantly decreased in the conditions of 30 min ischemia followed by different times of reperfusion. The releases of LDH and CK in coronary effluent were significantly increased in the conditions of 30 min ischemia followed by different times of reperfusion. Except the 10 min and 20 min ischemia, the releases of LDH and CK were significantly increased in Ovx during I/R. Ovx + E2 could abate the heart injury through decreasing the releases of LDH and CK. Besides the control and the 10 min I + 30 min R groups, the myocardial basal and ISO stimulation contraction were higher from Ovx than Sham, and the effect was reversed by Ovx + Ez.</p><p><b>CONCLUSION</b>The results indicate estrogen plays a cardioprotective role in global myocardial ischemia/reperfusion injury in ovariectomized (Ovx) rats.</p>
Subject(s)
Animals , Female , Rats , Estradiol , Pharmacology , Estrogens , Pharmacology , Myocardial Contraction , Physiology , Myocardial Ischemia , Myocardial Reperfusion Injury , Myocytes, Cardiac , Metabolism , Physiology , Ovariectomy , Random Allocation , Rats, Sprague-DawleyABSTRACT
<p><b>OBJECTIVE</b>To express the Gag protein of HIV-1 strain CN54 in Pichia pastoris (P.pastoris), optimize fermentation parameters and purify Gag antigen.</p><p><b>METHODS</b>The Gag gene was subcloned into downstream of aox1 promoter of Pichia expression vector pPS1.0, an integrative vector which possesses an identical 5' untranslated region as the natural aox1 gene and employs both in vitro construction and in vivo selection for multi-copy integrants. The recombinant vector was introduced into P.pastoris strain GS115 by electroporation and selected with G418 for Gag gene integration. Super G418 resistant clones were selected and screened for Gag expression. The engineered P.pastoris was cultured to high cell density (>300 A600 Units/ml) in a 5L fermentor. Through methanol induction, the expression level of Gag reached 120 mg/L. Intracellularly expressed Gag was released by high-pressure homogenization and purified through Sepharose FF and DEAE Sepharose FF column chromatography, the purity of Gag reached up to 90%.</p><p><b>RESULTS</b>Western-blotting suggested that purified Gag expressed in P.pastoris could react specifically with serum from HIV infected individual.</p><p><b>CONCLUSION</b>Gag antigen expressed in P.pastoris has provided a good basis for the development of a new generation of HIV vaccine candidates against some Chinese prevalent strains.</p>