Effect Comparison And Anlysis Of Mechanism Of Two New Kinds Of Inhibin DNA Vaccine On Rat

Two fusion expression plasmids of two copies inhibin and HBsAg gene (pCISI and pEGISI) were applied to immunize rats, through analysis on effect of immune original type, immune dosage; immune times to immune response, follicular development, the hormone levels and the reproduction and reproductive endocrinology, to compare the effect of two kind of inhibin gene vaccines, and to discuss the mechanism of inhibit gene vaccine.1 Immunoreactive character and affecting factors of inhibin gene immunizationA total of 90 rats were divided randomly into 9 groups (n=10) including six experimental groups. 100 μl procaine hydrochloride (0.5%) were injected as the pretreatment at 24 h before the immunization. In the group pCISI and pEGISI , three different dosages( 10, 50 and 100 μg/100 μ L) for each animal were respectively injected at the same site as the pretreatment. The control groups were injected with 100μl saline, 100 μg/100μL vectors pcMV-S or pEGFP-N1. A booster was performed as the primary immunization after 4 weeks.The results are as follows: ratio of positive antibody against INH for six experimental groups ranged from 10% to 50.0%. The highest value of P/N was determined to be 8.98. After twice immunizations of pCISI, 23.30%(7/30) of rats were detected in positive antibody against inhibin. The average of the value of antibody P/N value of 100 μg immunization group were remarkably higher than that of 10 μ g and 50 μ g on 4 weeks after booster. The increase of immunization dosage didn't promote the ratio of rats with positive antibody. After twice immunizations of pEGISI, 20.0%(6/30) of rats were detected in positive antibody against inhibin. The increase of immunization dosage raised the level of antibody. Considering the antibody level of immunized rats and positive rats, we found that, with regard to immunization with pCISI, rats immunized with 50 μ g produced the highest level of antibody against inhibin; with regard to immunization with pEGISI, rats immunized with 100 μ g produced the highest level of antibody against inhibin. The increase of immunization dosage wasn't sure to raise the ratio of rats with positive antibody.2 Effects of inhibin gene immunization on the reproduction and reproductive endocrinology of rats.The number of matured follicular(MF) in the 100 μ g group was 6.4 more than that in the control(p<0.05) after twice immunizations of pCISI, and there were 4.3, 6.4and 5.8more MF in 10, 50 ,100 μ g group after twice times immunizations of pEGISI than that in the control(p<0.05). 100μg pEGISI stimulated the follicular development obviously after twice immunizations (p<0.05). There is not a remarkable difference in plasma FSH between experimental and control rats on 4 weeks after booster of pCISI(p>0.05). After immunization of pEGISI, the level of FSH on estrus in 50 ,100 μ g groups was higher than that in the controls ,and remarkably more than 10 μ g group (p<0.05). The plasma 17-β estradiol went up after twice immunizations of pCISI. It went up after twice immunizations of pEGISI (p<0.05) on 2 weeks after booster of pEGISI, 50 μ g, 100 μ g pEGISI stimulated remarkably higher level of 17- β estradiol than the control group(p<0.05). The level of 17-β estradiol on estrus after twice immunization of pCISI, but there was no difference between positive and negative rats. The plasma progesterone went up after twice immunizations of pCISI. In conclusions, inhibin gene immunization can stimulate follicular development of rats. Inhibin gene immunization can raise the level of plasma FSH,17-β estradiol and progesterone, which set up the basis of inhibin gene immunizations on large animals.