Research On The Laboratory Efficacy And Safety Of Gene Adjuvant IL2 And IL-4 Of Chicken Coccidiosis Live Vaccine

In order to study the immune enhancement effect and safety of the gene adjuvant IL-2 and IL-4 of chicken coccidiosis live vaccine,which provides a scientific evidence for clinical application,p CI-ch IL-4-EGFP,p CI-ch IL-2-EGFP and p CI-ch IL-4-ch IL-2-EGFP were combined with chicken coccidiosis trivalent and four live vaccines to immunize SPF chicks,laboratory safety and distribution of expression in vivo were studied as well.The results showed:(1)All of the adjuvant group could make the vaccine efficacy test standard;the ACI and RWG increased as the increase of the dose,which decreased gradually when the dose was more than 250 ?g.The ACI made the high efficient when the does was 100 ?g p CI-ch IL-4-ch IL-2-EGFP,50 ?g p CI-ch IL-2-EGFP and 200 ?g p CI-ch IL-4-EGFP were mixed.It indicates that the optimal dose of adjuvant was 100 ?g p CI-ch IL-4-ch IL-2-EGFP,50 ?g p CI-ch IL-2-EGFP and 200 ?g p CI-ch IL-4-EGFP mixed.(2)p CI-ch IL-4-ch IL-2-EGFP(100 ?g)were intramuscularly inoculated with the coccidiosis vaccine concurrently,the interval between the two immunizations was 4~7 d.The results showed that it's could make the efficacy test standard when the adjuvant interval of the adjuvant group ?6 d,immune control group ?7 d.It is indicated that 100 ?g p CI-ch IL-4-ch IL-2-EGFP shortened the immune interval of live coccidiosis vaccine by 1 d.(3)The oral group(100~300 ?g)appeared an increasing trend as the dose increased.The ACI was188.41 when the does of oral administration was 300 ?g,which was close to the intramuscular group ACI value of 190.49.It is indicated that the immune effect of the oral group was effective,but oral administration dose is 3 times that of intramuscular injection.(4)It's could make the efficacy test standard when the adjuvant group 5th day and beyond,immune control group 6th day and beyond after the second immunization.It is indicates that 100 ?g p CI-ch IL-4-ch IL-2-EGFP can make the immune generation period shortened by 1d.(5)Chickens were given a single-dose and repeated single-dose intramuscularly with 100 ?g p CI-ch IL-4-ch IL-2-EGFP,and 1000 ?g p CI-ch IL-4-ch IL-2-EGFP was overdose.The results showed that there was no death in each group;the body temperature of the chickens was between 39.6~40.7?,whichwas consistent with the normal body temperature of healthy chickens;the average weight gain of the test group was not significantly different from that of the blank group(p>0.05).There was no lesions in each group of chickens,and no lesions were seen by HE staining either,genetic adjuvant does not transfer and integrate in microorganisms in the body.It indicates that the safety of the adjuvant is in line with the requirements of veterinary biological products.(6)SPF chickens were inoculated intramuscularly and orally with 100 ?g p CI-ch IL-4-ch IL-2-EGFP,the inoculation site was always the first to detect the plasmid;the second was the blood and heart;the liver,spleen,kidney and lung were at the end.The intramuscular route could be present for up to 21 d at the site of the inoculation site,it could be continuously expressed from 8 h after injection to 14 d after administration.It is indicated that the gene adjuvant could be continuously presented in vivo for long-term inoculation by intramuscular injection to 21 d.And the adjuvant is distributed and expressed in the body by the intramuscular route for a longer time than the oral route.