Effects Of Different Immunization On The Development Of Immune Organs,cell Proliferation And Expression Of Apoptosis-relatived Gene In Chicken

The experiment was conducted to determine the stress of different immunizations on Gushi chicken and effects on the development of immune organs,cell cycle,cell proliferation and differentiation and expression level of apoptosis-related gene.Experimental design240 one-day-old healthy Gushi male chickens were randomly assigned to 4 groups,each group contained 4 pens and 15 chickens per pen.Group A was control group without any immune, group B was given intranasal-conjunctival immunization, group C and D were given both intranasal-conjunctival immunization and subcutaneous-injection immunization. Additionally, vitamin E (100IU/kg) was added into diet of group D from 7-day-old. Chicken were treated with different immunizations at 10 days.Trail 1 Effects of different immunizations on stress-related hermone level in chickenThree chickens of each pen were randomly caught and blood samples were collected after 1,3,5,and 7d of immuned, serum concentration of CORT and ACTH were measured.The results suggested that:1day after immunization, serum concentration of CORT in group D was highest and significantly higher than group A and B(p<0.05);during 3 to 7days after immunization,CORT concentration of all group decreased to a similar level,while group D kept in a lower level;ACTH concentration of all groups had no significant difference(p>0.05).It indicated that subcutaneous-injection immunization caused severe stress on chicken ,and it was down-regulated by VE added in diet.Trail 2 Effects of different immunizations on body weights and the development of immune organs in chickenThree chickens of each pen were randomly caught and weighted before killed,immune organs were collected and weighted,organ index were calculated. The results suggested that:body weights had no significant difference within each group(p>0.05);Thymus index of group C was significantly higher than other groups at 3,5 and 7 days after immunization(p<0.05),while spleen index and bursa index had no significant difference in all treatments.These results indicated that: different immunizations had no significant effect on body weights, subcutaneous-injection immunization caused higher development of thymus,while had no effects on spleen and brusa of chicken.Trail 3 Effects of different immunizations on cell proliferation of immune organs in chickenOne chickens of each pen were randomly caught and weighted before killed,immune organs were collected for cell cycle assay by FCM. The results suggested that:it had significant effect on S phase and G2/M phase by different immunizations, cell proliferation indexes of thymus and brusa were down-regulated,while cell proliferation indexes of spleen had no effect.Trail 4 Effects of different immunizations on relative expression level of apoptosis-related gene of immune organs in chickenOne chickens of each pen were randomly caught and weighted before killed,immune organs were collected,total RNA were detection and reverse transcription into cDNA,SYBR GreenⅠreal-time PCR were used to test expression level of Bcl-2,Fas,andFasL.The results showed that expression of Bcl-2 and FasL in thymus were higher by subcutaneous-injection immunization; Bcl-2 gene was lower by different immunization,while expression of Fas was higher in spleen; Relative expression level of Bcl-2 was higher in bursa at 5d after immunization,and relative expression level of Fas and FasL were also higher in bursa at 3-5d after immunization.