Research On Efficacy Of Immunocontraception By Using Human Urokinase To Immunize Male Mice

PARTⅠMethods of Immunocontraception by Using Human Urokinase to Immunize Male Mice and the Potency AnalysisObjective: To establish a method of inducing Kunming male mice to produce human uPA-specific antibody, and to detect the efficacy and stability of the method.Methods: Thirty adult Kunming male mice were randomly divided into five groups: blank control group, adjuvant control group(100μl Physiological saline+100μl adjuvant), uPA low dose immunized group(20μg/100μl uPA solution+100μl adjuvant), uPA middle dose immunized group(40μg/100μl uPA solution+100μl adjuvant) and uPA high dose immunized group(80μg/100μl uPA solution+100μl adjuvant). The antigen mixed with Freund's adjuvant was subcutaneous injected once every two weeks. In one week after the third immunity, Antibody potency of every group were determined by enzyme linked immunosorbent assay(ELISA).Results: As compared with control groups, the antibody content of human uPA-specific antibody in uPA three-doses immunized groups significantly increased(P<0.05);with the increase in immunization dose the content of antibody showed a downward trend, but had no statistically significant difference(P>0.05) among these groups; the potency of anti-uPA polyclonal antibody in immune serum all reached 1:10240 or higher levels.Conclusion: Using human uPA antigen to immunize by there times, mice of the uPA immunized groups could produce a stable antibody potency.PARTⅡThe Effects of Reproductive Capacity on Male Mice after Immunizing by Human UrokinaseExperiment 1 The Impacts of Mating and Fertility on Male Mice after Immunizing by Human UrokinaseObjective: To observe the changes in mating and fertility on male mice after immunized by human urokinase.Methods: Animal groups and immunization were the same as partⅠ. After the third immunity, male mice were coupled with the adult female mice. The vaginal embolus or sperm in vaginal cells smear was inspected in next morning, and the mating rate was caculated. 14 days after the successful mating, females were sacrified to observe the pregnancy and the number of live embryos in pregnant mice of every groups.Results: There is no significant difference in the mating rate between the immunized groups and the control groups. Pregnancy rates and the average number of live embryos of the three doses uPA immunized groups markedly decreased(P<0.001), there was no significant difference between blank control and adjuvant control(P>0.005).Conclusion: Using human uPA to immunize male mice there was no significant change in mating but their fertility decline. Experiment 2 The Impacts of Sperm Function on Male Mice after Immunizing by Human UrokinaseObjective: To observe the changes in sperm function on male mice after inmmunizing by human uPA.Methods: Animal grouping and immunization refer to partⅠ. After the third immunity, the male gonadal organs were weighted, and the sperm function (such as: motile rate, viable rate, swollen rate, sperm density, activity, sperm chemotaxis that induced by the uPA, in vitro fertilization rate) of every groups were evaluated.Results:①Gonadal organ indexes of each group had no significant difference(P>0.05);②Sperm survival rate: compared with the control groups, sperm survival rates of three doses uPA immunized groups had a significant decline(P<0.05);③Sperm motile rate: compared with the control groups, sperm motile rate of three doses uPA immunized groups had a significant decline(P<0.05);④Sperm swollen rate: compared with the control groups, sperm swollen rates of uPA low dose immunized group had a significant increase(P<0.05), that of other doses uPA groups had no significant difference(P>0.05);⑤Sperm density: compared with the control groups, sperm density of three doses uPA immunized groups had a significant decline(P<0.05);⑥Sperm run-up experiment: there was no significant difference in run-up distances between every groups(P>0.05);⑦Sperm chemotaxis that induced by the uPA: compared with the control groups, sperm chemotaxis that induced by the uPA of uPA middle and high doses immunized groups had a significant decline(P<0.05), that of uPA low dose immunized group had no significant difference(P>0.05);⑧In vitro fertilization rate: compared with the control groups, in vitro fertilization rates of three doses uPA immunized groups had a significant decline(P<0.05).Conclusion: There was significant decline in sperm function on male mice after immunzing the male mice by human uPA.