1.Using mimicry epitopes from phage displayed peptide library as substitute antigen for cyclophilin A in ELISA for detection of anti-CyPA autoantibody
Chinese Journal of Laboratory Medicine 2003;0(08):-
Objective To use mimicry epitopes from phage displayed peptide library as substitute for recombinant human cyclophilin A (rhCyPA) in detection of CyPA autoantibody in sera of patients with systemic lupus erythematosus (SLE).Methods Monoclonal antibody D4 and E6 were used as the capture antibody; the Ph.D.-12 p hage display peptide library was screened by biopanning. Ten phage clones with h igh affinity for McAb D4 and E6 (five for each) were chosen. Their inserts were sequenced. The amino sequence of mimicry epitope was subsequently reasoned out. Selected 12 peptide and previously obtained 7 peptide mimicry epitopes were used as coated antigen and compared with rhCyPA in ELISA for anti-CyPA autoantibody .Results Nine of ten phage clones with high affinity for McAb D4 and E6 (five for D4, four for E6) shared a consensus amino sequence HW EYTTSPHPRL. In comparative study of ELISA, 56 serum samples from SLE patients we re tested for anti-CyPA autoantibody, 28 samples were positive (50%) with 12 pe ptide mimicry antigen and 24 were positive (43%) with rhCyPA, 4 of 34 (11%) samp les from those with other autoimmune disorders were positive with both antigens, the coincidence rate between two methods was 95 6%. The research also showed t hat 12 peptide mimicry antigen has the advantages of rhCyPA and 7 peptide mimicr y antigen in detecting anti-CyPA autoantibody. Conclusions The specific 12 peptide screened from phage-displayed peptide library by ant i-CyPA monoclonal antibody can mimic epitope information of cyclophilin A well and be used as substitute for rhCyPA in ELISA.
2.Effect of olmesartan medoxomil on the expression of osteoprotegerin in rats with atherosclerosis
Guiming CHEN ; Zhe SHEN ; Zhengzhang LI ; Yefu SUN ; Jiagao QIU ; Fuyin LUO
International Journal of Laboratory Medicine 2017;38(17):2374-2376
Objective To investigate the effect of olmesartan medoxomil on osteoprotegerin and serum inflammatory factor expression in atherosclerosis model rat.Methods From 2015 April in Jiangsu Institute of schistosomiasis control,24 Sprague-Dawley rats were randomly divided into the following three groups:the control group was fed with standard food,the model group was fed with high fat diet based on standard food(normal food +2% cholesterol +5% goat fat +0.2% acid +7×105 IU/(kg·d) Vitamin D3),and the olmesartan group was given 3 mg/(kg·d) olmesartan gavage daily on the basis of high fat diet.On the fourth week of experiment the level of serum high sensitivity C reactive protein and serum lipids include total cholesterol,low density lipoprotein,high density lipoprotein and triglyceride were detected.The structure and changes of aortic pathology was observed.The expression of osteoprotegerinin in aortic was detected by immunohistochemistry staining and Western blot.Results Within the model group,the level of serum lipid and high-sensitivity C reactive protein increased significantly,endometrial thickness,intima-to-media thickness ratio and osteoprotegerin expression in aorta was significantly higher than that of normal group,the difference was statistically significant(P<0.05).The serum high density lipoprotein increased significantly,the level of other serum lipids and high sensitive C reaction protein decreased significantly,endometrial thickness,intima-to-media thickness ratio and osteoprotegerin expression in aorta of the olmesartan treated rats were significantly lower than those of model animals,the difference was statistically significant(P<0.05).Conclusion Olmesartan may suppress the development of atherosclerosis in model rats by decreasing the expression of osteoprotegerin and the level of serum inflammatory cytokines.