Interleukin-8 homolog (vIL-8) is a chemokine encoded by the genome of Marek's disease virus. Chicken IL-8 (cIL8) is an important chemokine of chickens which plays a role in antiviral activity. To explore the relat...Interleukin-8 homolog (vIL-8) is a chemokine encoded by the genome of Marek's disease virus. Chicken IL-8 (cIL8) is an important chemokine of chickens which plays a role in antiviral activity. To explore the relationship between vIL-8 and cIL8 can help to discern the function of vIL-8. In this study, the amino acid sequences of vIL-8 and cIL8 were aligned. The cIL8 gene was expressed in E. coli and the cIL8 fusion protein was obtained, after induction with IPTG. The protein was separated by SDS-PAGE and the band of interest was excised and minced for mouse immunization. An immunofluorescence test was used to detect vIL-8 expression in insect cells. The results showed that both vIL-8 and cIL8 were typical CXC chemokines in structure, and they had similar key amino acids, known to be important for receptor binding. The result of the immunofluorescence test showed that the mouse anti-cIL8 serum could react with vIL-8 expressed by insect cells. Therefore, vIL-8 shares common antigenic determinants with cIL8, and they may have a common receptor. This feature of vIL-8 suggests that it may participate in the immune evasion of the virus.展开更多
Five to ten serotyqe I Marek’s disease virus (MDV1) strains of different pathotypes were compared for their DNA sequences of gI, gE, pp38 and meq genes. The reference strains were vMDV GA and JM, vvMDV RB1B and Md11(...Five to ten serotyqe I Marek’s disease virus (MDV1) strains of different pathotypes were compared for their DNA sequences of gI, gE, pp38 and meq genes. The reference strains were vMDV GA and JM, vvMDV RB1B and Md11(p16), vv+MDV strains 648A and 584A,vaccine strain CVI988/Rispens; Chinese strains were: v MDV strain N, vvMDV strain G2, vaccine strain 814. Only random aa changes were found in 12 positions within gE of 497 aa among 10 analyzed strains and in 10 positions within gI of 355 aa among 5 strains. There was no relationship found between virus pathotypes and aa changes of both glycoproteins. The aa changes happened in 14 positions within meq of 339 aa among 5 compared strains. But a proline deletion at aa #194 in a proline-rich domain of meq were shared by two vaccine strains CVI988/Rispens and 814, the latter was a non-pathogenic vaccine strain of serotype 1 isolated in 1983 in China. In another hand, vv+MDV strains 648A demonstrated 5 unique aa changes and 4 of them also located in the proline-rich region. The pp38 was very conservative among sequenced 10 strains, there were only two positions at #107 and #109 with aa altered in its 290 aa. All tested MDV1 strains had glutamine at #107, instead, only vaccine CVI988 had arginine at the position and lost its epitope reactive with Mab H19, to which all other tested MDV1 strains were positive. However, CVI988 and vMDV GA shared a Mab T65-recognized epitope when there was glycine at aa#109. It was glutamic acid at aa#109 in all other MDV1 strains which were not reactive with Mab T65. It seems like that there is some relationship between pathotypes and sequences of pp38 and meq, but more strains need to be compared.展开更多
基金funded by Program for Changjiang Scholars and Innovative Research Team in University(IRT0978,PCSIRT)the National Natural Science Foundation of China(30270982)
文摘Interleukin-8 homolog (vIL-8) is a chemokine encoded by the genome of Marek's disease virus. Chicken IL-8 (cIL8) is an important chemokine of chickens which plays a role in antiviral activity. To explore the relationship between vIL-8 and cIL8 can help to discern the function of vIL-8. In this study, the amino acid sequences of vIL-8 and cIL8 were aligned. The cIL8 gene was expressed in E. coli and the cIL8 fusion protein was obtained, after induction with IPTG. The protein was separated by SDS-PAGE and the band of interest was excised and minced for mouse immunization. An immunofluorescence test was used to detect vIL-8 expression in insect cells. The results showed that both vIL-8 and cIL8 were typical CXC chemokines in structure, and they had similar key amino acids, known to be important for receptor binding. The result of the immunofluorescence test showed that the mouse anti-cIL8 serum could react with vIL-8 expressed by insect cells. Therefore, vIL-8 shares common antigenic determinants with cIL8, and they may have a common receptor. This feature of vIL-8 suggests that it may participate in the immune evasion of the virus.
文摘Five to ten serotyqe I Marek’s disease virus (MDV1) strains of different pathotypes were compared for their DNA sequences of gI, gE, pp38 and meq genes. The reference strains were vMDV GA and JM, vvMDV RB1B and Md11(p16), vv+MDV strains 648A and 584A,vaccine strain CVI988/Rispens; Chinese strains were: v MDV strain N, vvMDV strain G2, vaccine strain 814. Only random aa changes were found in 12 positions within gE of 497 aa among 10 analyzed strains and in 10 positions within gI of 355 aa among 5 strains. There was no relationship found between virus pathotypes and aa changes of both glycoproteins. The aa changes happened in 14 positions within meq of 339 aa among 5 compared strains. But a proline deletion at aa #194 in a proline-rich domain of meq were shared by two vaccine strains CVI988/Rispens and 814, the latter was a non-pathogenic vaccine strain of serotype 1 isolated in 1983 in China. In another hand, vv+MDV strains 648A demonstrated 5 unique aa changes and 4 of them also located in the proline-rich region. The pp38 was very conservative among sequenced 10 strains, there were only two positions at #107 and #109 with aa altered in its 290 aa. All tested MDV1 strains had glutamine at #107, instead, only vaccine CVI988 had arginine at the position and lost its epitope reactive with Mab H19, to which all other tested MDV1 strains were positive. However, CVI988 and vMDV GA shared a Mab T65-recognized epitope when there was glycine at aa#109. It was glutamic acid at aa#109 in all other MDV1 strains which were not reactive with Mab T65. It seems like that there is some relationship between pathotypes and sequences of pp38 and meq, but more strains need to be compared.
