ABSTRACT
UL48 plays essential role in replication of MDV genome and interacts with UL36 as well as other MDV tegument proteins.To investigate the interaction between UL48 and UL36 during MDV oncogenisis,antibody against UL48 was prepared and characterized in current study.UL48 gene was amplified from MDV-Ⅰ genome and then subcloned into pTYB1 and pGEX-4T3 vectors for UL48 expression with induction of IPTG in BL21(DE3) E..coli cells.Chitin-sepharose and Glutathion-sepharose were,respectively,used to purify fusion protein intein-UL48 and GST-UL48.Four subcutaneous injections of intein-UL48 fusion protein were done on the lower back and the thigh of rabbit and then other three injections with an interval 10 days.The titer of antibody was measured by the sandwich ELISA with UL48 protein isolated from GST-UL48 after cleavage of thrombin.Western blot was carried out for specificity analysis of antibody against UL48 protein.The results suggested that UL48 antibody was succesfully prepared,and its titer was 1 ∶ 512 000.
ABSTRACT
Objective To investigate the expression of blood CD +4 CD +25 Treg GITR,CD +4 T cell GITRL in children with asthma,and the role of them in asthmatic inflammation.Methods 50 cases of severe asthma were selected,and were controlled with thirty two healthy children.The venous blood was collected both in the period of acute episode and clinic remission.The mean fluorescence intensity of CD +4 CD +25 Treg GITR and CD +4 T cell GITRL was detected by flow cytometry.Results The expression of CD +4 CD +25 Treg GITR in the asthma acute period group was (24.2 ±8.2)MFI,which was significantly lower than (28.5 ±6.0)MFI in the control group(t =2.5,P 0.05).Moreover,the expression of CD +4 CD +25 Treg GITR in the asthma in remission group after treatment was (29.5 ±8.3)MFI,which was significantly higher than that in acute period group before treatment(t =-9.9,P 0.05).Furthermore,there was no significant correlation between levels of CD +4 CD +25 Treg GITR and CD +4 T cell GITRL.Conclusion The level of CD +4 CD +25 Treg GITR in acute period asthmatic patients was decreased,but it was increased in remission,but no changes of CD +4 T cell GITRL expression were observed.GITR/GITRL signal system might be involved in the asthmatic inflammation procession.