ABSTRACT
Objective:To build murine asthma model with Dermatophagoides pteronyssinus. To compare the curative effects of the immunotherapy regimens with either Dermatophagoides pteronyssinus allergen extracts or a recombinant Derp2(rDerp2). To offer a novel thread for immunotherapy of allergic diseases.Methods:The BALB/c mice were divided into four groups randomly. They were: negative control, positive control, allergen extracts and rDerp2. Mice were sensitized by i.p. injection of mixture of allergen extracts and Al(OH)3 except the negative control,and were i.p. given a treatment with either allergen extracts or rDerp2 respectively, and then were i.n. given allergen extracts as a challenge. Negative control was treated by saline always. Assays for IgE,IgG1,IgG2a in serum,IL-4,IFN-? in BALF and SCCS were performed by ELISA. Total cell number and assorted cell number in BALF samples were determined. The lungs kept in 10% formalin were made into slices stained by HE in order to observe the pathologic changes in lungs.Results:Compared to the negative control, in the positive control the total cell number and the number of inflammatory cell in BALF were significantly increased, BALF eosinophil counts were significantly increased, neutrophils and eosinophils were the predominant BALF inflammatory; and the Th2 cytokine, e.g. IL-4, was the main cytokine in BALF and SCCS. Compared with the positive control, the allergic responses in the mice given relevant immunotherapy were obviously alleviated, and the predominant cytokine in BALF and SCCS was Th1 pattern, e.g. IFN-?.Conclusion:We established successfully the murine asthma model. Immunotherapy with Dermatophagoides pteronyssinus allergen extracts had better curative effects than immunotherapy with recombinant Derp2.
ABSTRACT
Objective To investigate the efficacy and mechanism of subcutaneously given recombinant Der p 2 entrapped PLGA nanoparticles(DEPN) on mouse model with allergic airway inflammation.Methods 40 BALB/c mice were randomly divided into 5 groups,group A(normal control) were treated with saline(100 ?l) all the time,groups B,C,D and E were sensitized intraperitoneally with crude dust mite extracts(10 ?g) and then subcutaneously treated respectively with PBS(100 ?l),2 mg empty PLGA(EP),100 ?g rDer p 2,and 2 mg DEPN(loaded with 100 ?g rDer p 2) for 3 times,once per day,followed by intranasal challenge of 50 ?g rDer p 2.One day post challenge,mice were sacrificed and bronchoalveolar lavage fluid(BALF) was collected.Number of the total cells and eosinophils was determined,and airway inflammation and mucus secretion were analyzed by haematoxylin and eosin(H&E) staining and periodic acid-Schiff(PAS) staining.Level of cytokines in the supernatant of splenocyte culture was assayed by ELISA.Level of rDer p 2 specific IgG2a and IgE in the sera was determined by ELISA.Results The lung histology showed development of eosinophil infiltration in the airway of mice in groups B and C.The lung inflammation and mucus secretion in groups D and E were significantly alleviated than that of groups B and C.Number of total cells(63.50?5.12) andeosinophils(15.32?3.04) in BALF decreased in group B.Compared with group B,the number of total cells in groups D(55.3?5.20) ? 104 /ml and E(41.00?4.91) ?104 /ml greatly decreased(P