[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan ...[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan three-way cross-breeding pigs. According to the porcine IL-18 gene in GenBank, a pair of specific primers was designed. The full length cDNA of porcine IL-18 was amplified by RT-PCR. Subsequently, porcine IL-18 cDNA was cloned into pGEM-T vector and sequenced and analyzed. [ Result] The porcine IL-18 gene demonstrated an open reading frame of 579 bp encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and cleaved by interleukin-1 beta （IL-1β） converting enzyme（ICE） in caspase-1 splice site; the porcine mature protein had biological activity： After comparing with other porcine IL-18 genes, the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [ Conclusion] A full length procine IL-18 gene was gained. It lays the foundation for porcine IL-18 as an adjuvant of genetic vaccine.展开更多
The purpose of this investigation was to evaluate the effect of a dietary supplement （Mirigen TM）, a fungal cell wall derivative product, as a new generation alternative to antibiotics, on the growth and the innate ...The purpose of this investigation was to evaluate the effect of a dietary supplement （Mirigen TM）, a fungal cell wall derivative product, as a new generation alternative to antibiotics, on the growth and the innate and adaptive immune functions in broilers from birth to 45 d of age. Newborn chicken were randomly assigned to one of three groups： G 1 （n=150） controls no supplement fed; G2 （n=150） is fed with dietary supplement at a designed regular dose （0.5 %, weight of additive to food）; G3 （n= 150） is fed with dietary supplement at double doses （1%）. All three groups were housed in the same conditions. Body weight and blood were taken on day 1, 14, 28 and 45. Medications used and costs/treatment were recorded for each group. The whole blood was used to purify heterophils for reactive oxygen species （ROS） generation and E coli killing abilities examination assays, and the serum samples were preserved in freezer for enzyme linked immunosorbent assays （ELISAs） to determine concentration of macrophage inflammatory protein-1β （MIP-1β）, CD4/CD8, interferon-y （IFN-y）, and titers of antibody against Newcastle disease virus （NDV）. Group differences were analyzed using analysis of variance （ANOVA） algorithm （S-Plus）. There was no significant birth weight difference in three groups. After 45 d growth, the dietary supplement treated groups had significantly higher body weight gain （BWG） with lower mortality rate if compared to the untreated control group （P〈0.05）. Their BWG and mortality rate were 2.23 kg and 10 % in GI （control group）, 2.89 kg and 2 % in G2 （experimental group, 0.5 % dose）, and 2.77 kg and 1% in G3 （experimental group, 1% dose）, respectively. Heterophil ROS generation in treated groups were markedly improved through the addition of dietary supplement in both regular and double doses to the diet （P〈 0.05）. The ability of heterophil to kill E coli was also significantly improved in dietary supplement treated groups （P〈0.01）. Comparing to control group, there was significantly higher serum IFN-y concentration in treated groups （P〈0.05） on day 45. The CD4/CD8 was also improved in treated groups （P〈0.05）. Newcastle di-sease is the most prevalent avian disease, and vaccination is an effective method to protect the animals from the virus infection. In our study, it is found G2 and G3 that fed with dietary supplement had higher antibody titers against NDV after vaccination （P〈0.01） and the antibodies lasted longer. Results from this study demonstrated dietary supplement to broilers improved the immune capabilities of immune cells, which are vital to the establishment of immune response against pathogens, thereby, to improve chicken＇ s health and growth and reduce medication cost in chicken farming.展开更多
基金Supported by Project of Basic and Frontier Technology Research in Henan Province(072300430060 )Key Scientific and Technological Project of Henan Province(072102130023)~~
文摘[ Objective] To clone the porcine interleukin-18（/L-18） cDNA and explore the immunological effectiveness of porcine IL-18 as an adjuvant of genetic vaccine. [ Method] The spleen lymphccytes were isolated from Henan three-way cross-breeding pigs. According to the porcine IL-18 gene in GenBank, a pair of specific primers was designed. The full length cDNA of porcine IL-18 was amplified by RT-PCR. Subsequently, porcine IL-18 cDNA was cloned into pGEM-T vector and sequenced and analyzed. [ Result] The porcine IL-18 gene demonstrated an open reading frame of 579 bp encoding an inactive precursor protein with 192 amino acids. The precursor protein had no typical hydrophobic signal peptide and cleaved by interleukin-1 beta （IL-1β） converting enzyme（ICE） in caspase-1 splice site; the porcine mature protein had biological activity： After comparing with other porcine IL-18 genes, the nucleotide sequence homology was over 96% and the deduced amino acid homology was more than 98%. [ Conclusion] A full length procine IL-18 gene was gained. It lays the foundation for porcine IL-18 as an adjuvant of genetic vaccine.
文摘The purpose of this investigation was to evaluate the effect of a dietary supplement （Mirigen TM）, a fungal cell wall derivative product, as a new generation alternative to antibiotics, on the growth and the innate and adaptive immune functions in broilers from birth to 45 d of age. Newborn chicken were randomly assigned to one of three groups： G 1 （n=150） controls no supplement fed; G2 （n=150） is fed with dietary supplement at a designed regular dose （0.5 %, weight of additive to food）; G3 （n= 150） is fed with dietary supplement at double doses （1%）. All three groups were housed in the same conditions. Body weight and blood were taken on day 1, 14, 28 and 45. Medications used and costs/treatment were recorded for each group. The whole blood was used to purify heterophils for reactive oxygen species （ROS） generation and E coli killing abilities examination assays, and the serum samples were preserved in freezer for enzyme linked immunosorbent assays （ELISAs） to determine concentration of macrophage inflammatory protein-1β （MIP-1β）, CD4/CD8, interferon-y （IFN-y）, and titers of antibody against Newcastle disease virus （NDV）. Group differences were analyzed using analysis of variance （ANOVA） algorithm （S-Plus）. There was no significant birth weight difference in three groups. After 45 d growth, the dietary supplement treated groups had significantly higher body weight gain （BWG） with lower mortality rate if compared to the untreated control group （P〈0.05）. Their BWG and mortality rate were 2.23 kg and 10 % in GI （control group）, 2.89 kg and 2 % in G2 （experimental group, 0.5 % dose）, and 2.77 kg and 1% in G3 （experimental group, 1% dose）, respectively. Heterophil ROS generation in treated groups were markedly improved through the addition of dietary supplement in both regular and double doses to the diet （P〈 0.05）. The ability of heterophil to kill E coli was also significantly improved in dietary supplement treated groups （P〈0.01）. Comparing to control group, there was significantly higher serum IFN-y concentration in treated groups （P〈0.05） on day 45. The CD4/CD8 was also improved in treated groups （P〈0.05）. Newcastle di-sease is the most prevalent avian disease, and vaccination is an effective method to protect the animals from the virus infection. In our study, it is found G2 and G3 that fed with dietary supplement had higher antibody titers against NDV after vaccination （P〈0.01） and the antibodies lasted longer. Results from this study demonstrated dietary supplement to broilers improved the immune capabilities of immune cells, which are vital to the establishment of immune response against pathogens, thereby, to improve chicken＇ s health and growth and reduce medication cost in chicken farming.
